Please Scroll Down to See Forums Below I've done well on a single 20mg tablet per day.. Most guys continue Nolvadex for a few weeks anyway, so I don't think it a bad idea. Cholesterol levels do tend to rebound a little slowly..
- Bill Llewellyn
- Bill Llewellyn
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Winstrol is one of the safe anabolics,right?Wrong!!Actually it is one of the most dan
- Thread starter Realgains
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macrophage69alpha
New member
Lipoprotein effects are due to its effects on Hepatic lipase. Taking nolva with winstrol may help.. but Nolva will make YOU FAT(if you would like I will post the studies)... adding a SERM is ok, but clomid might be a better choice.
you would be better off stacking with test, even used with arimidex you will get some estrogen (which will not happen with winstrol)
NOTE- STACKING WITH DECA MAY EXACERBATE THE HDL PROBLEM... AND IT DECREASES THE BENEFICIAL EFFECTS OF ANY SERM OR NATURAL ESTROGEN PRESENT...
Hepatic lipase activity influences high density lipoprotein subclass distribution in normotriglyceridemic men: genetic and pharmacological evidence
Scott M. Grundy, Gloria L. Vega, James D. Otvos,
David L. Rainwater, and Jonathan C. Cohen
Abbreviations:
HDL-C, high density lipoprotein cholesterol;
GGE, gradient gel electrophoresis;
NMR, nuclear magnetic resonance.
Abstract: Several studies have reported an inverse relationship between hepatic lipase activity and plasma high density lipoprotein (HDL) cholesterol concentrations. The purpose of the present study was to determine whether genetic and pharmacological variation in hepatic lipase activity alters the distribution of HDL subclasses. Two independent analytical methods (nuclear magnetic resonance and gradient gel electrophoresis) were used to compare HDL subclass distributions in 11 homozygotes for the -514C allele of hepatic lipase and in 6 homozygotes for the -514T allele. Mean hepatic lipase activity was 45 ± 15 mmol× l-1× hr-1 in -514C homozygotes and 20 ± 7 mmol× l-1× hr-1 in -514T homozygotes. Both analytical methods indicated that HDL2b was significantly higher and HDL3a was significantly lower in -514T homozygotes than in -514C homozygotes. No differences were noted in the other HDL fractions (HDL2a, HDL3b, and HDL3c). To determine the effects of increased hepatic lipase activity, 20 men were given the synthetic anabolic steroid, stanozolol. Stanozolol treatment increased hepatic lipase activity more than two-fold (38 ± 18 to 85 ± 25 mmol× l-1× hr-1), and markedly reduced the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a). The plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were unchanged by stanozolol treatment. Taken together, these genetic and pharmacological data indicate that variation in hepatic lipase activity has highly specific effects on the distribution of HDL subclasses in the circulation. -Grundy, S. M., G. L. Vega, J. D. Otvos, D. L. Rainwater, and J. C. Cohen. Hepatic lipase activity influences high density lipoprotein subclass distribution in normotriglyceridemic men: genetic and pharmacological evidence. J. Lipid Res. 1999. 40: 229-234.
Human hepatic lipase is a 476 amino acid glycoprotein that exhibits phospholipase A1 and triglyceride hydrolase activities (1, 2). The enzyme is synthesized in hepatocytes, secreted, and bound to hepatocyte and hepatic endothelial surfaces (3), from which it can be released into the circulation by heparin. Data from several clinical, pharmacological, and genetic studies support an inverse relationship between hepatic lipase activity and the plasma concentrations of high density lipoprotein cholesterol (HDL-C) in humans. The activity of hepatic lipase in postheparin plasma appears to be inversely correlated with plasma HDL-C concentrations among unselected individuals (4-8). Drugs such as anabolic steroids that increase hepatic lipase activity decrease HDL concentrations (9), whereas hepatic lipase deficiency is associated with high plasma HDL levels in some patients (10). Studies of normolipidemic families indicated linkage between plasma HDL concentrations and LIPC, the gene encoding hepatic lipase (11, 12). DNA sequencing revealed an LIPC allele (designated -514T) containing four linked polymorphisms in the 5' flanking region. This allele was associated with low hepatic lipase activity (13) and increased plasma HDL-C concentrations in white men (12). These findings suggest that hepatic lipase plays a role in the catabolism of HDL, and that normal variation in hepatic lipase activity is associated with inter-individual differences in plasma HDL concentration.
Some studies have indicated that hepatic lipase activity is correlated with the plasma concentrations of specific HDL subfractions. Kuusi, Saarinen, and Nikkila (4) reported that hepatic lipase activity was inversely correlated with plasma concentrations of HDL2, but not HDL3 determined by ultracentrifugation. Katzel et al. (14) found that postheparin plasma hepatic lipase activity was inversely correlated with the proportion of HDL in the HDL2b subspecies determined by gradient gel electrophoresis (GGE) in 41 healthy men.Johansson et al. (15) reported that hepatic lipase was inversely correlated with HDL2b and positively correlated with HDL3b (determined by GGE) in 24 normolipidemic infarct survivors. These observations are consistent with in vitro studies indicating that hepatic lipase acts preferentially on large HDL particles (16), and suggest that hepatic lipase plays a role in the remodeling of circulating HDL. However, all of the clinical studies performed to date were based on correlations observed between two phenotypes, hepatic lipase activity and HDL subclass distribution. Such correlations are subject to confounding by secondary factors and do not constitute definitive evidence for a causal relationship between hepatic lipase activity and plasma HDL-C concentrations. For example, plasma triglyceride concentrations are positively correlated with hepatic lipase activity (5) and inversely related to plasma HDL-C concentrations ( 17-20) . Therefore a correlation between hepatic lipase activity and plasma HDL size among unselected individuals may simply reflect the interactions of plasma triglyceride concentrations (or some other confounder) with both parameters.
Secondary confounding can be avoided by studying individuals with genetic (i.e., primary) variation in hepatic lipase activity. Detailed analysis of HDL subclass distribution in such subjects is limited to a single report by Knudsen et al. (20), who studied a compound heterozygote for two hepatic lipase gene mutations. Hepatic lipase activity in this individual was decreased by 70% compared to healthy control men. Density gradient ultracentrifugation of his HDL revealed markedly increased concentrations of the IDLE and HDL2b subclasses. The present study was undertaken to determine whether variation in hepatic lipase activity alters the distribution of HDL subclasses. To minimize confounding by secondary factors, HDL subclass distributions were measured in normotriglyceridemic men with genetically defined differences in hepatic lipase activity. To ensure reliable estimation of HDL subclass distribution, two independent analytical methods (nuclear magnetic resonance and gradient gel electrophoresis) were used. To support the findings of this genetic approach, the effects of pharmacological stimulation of hepatic lipase activity on HDL size distribution was determined.
METHODS
Experimental design
Fasting blood samples were drawn from 21 normolipidemic white men. Six of the men were homozygous for the -514T allele of LIPC, four were -514C/ -514T heterozygotes, and 11 were -514C homozygotes. Five of the -514T homozygotes and five of the -514C homozygotes had been matched for plasma HDL-C concentrations in a previous study Aliquots of plasma were stored at 4°C for lipid and lipoprotein measurement, and at -80°C for HDL subclass analysis. Heparin injections for determination of postheparin plasma hepatic lipase activity were performed after an overnight fast. Twenty of the men then took 0.2 mg/kg per day of the oral synthetic anabolic steroid stanozolol (Winstrol, Winthrop, Pittsburg, PA) up to a maximum of 16 ma/ day for 6 consecutive days. Compliance was monitored by pill counting. The drug was well tolerated by all of the men, and no side effects were reported. A second heparin injection was performed within 24 h after ingestion of the final dose of stanozolol.
Subjects
The study was approved by the Institutional Review Board at the University of Texas Southwestern Medical Center. Men aged 20 to 50 years who were apparently healthy, who had plasma triglyceride concentrations below 200 mg/dL, and who did not smoke or use lipid-lowering drugs or beta blockers were recruited by advertisements placed on notice boards at Southwestern Medical Center. Subjects were studied as outpatients on their ad libitum diets.
Hepatic lipase genotyping
Hepatic lipase genotypes were determined by PCR amplification and restriction enzyme digestion as described in detail previously (12).
Assay of hepatic lipase activity
Hepatic lipase activity was measured in postheparin plasma using a modification of the method reported by Baginsky and Brown (22) as described in detail previously (8). Briefly, patients received an intravenous injection of 75 IU of porcine heparin (Elkins-Sinn Inc.) per kg of body weight after a 12-h fast. Blood samples were drawn into tubes containing EDTA before and 15 min after injection. Plasma was separated immediately upon collection of blood and aliquots were frozen at -70°C. Hepatic lipase activity was measured by addition of 0.025 mL postheparin plasma to 0.475 mL medium (pH 8.8) containing NaCI (0.75 m), a gum arable-stabilized emulsion of triolein (final concentration 15 mmol/L) labeled with [3H]triolein (0.5 m Ci), and fatty acid-free bovine serum albumin (final concentration 50 g/L). The free [3H]oleate generated during the hydrolysis reactions was extracted using the method of Belfrage and Vaughan (23). The intra- and inter-assay coefficients of variation were <5%.
Assay of plasma lipids and lipoproteins
Plasma triglyceride, cholesterol, and HDL-C concentrations were measured using standard enzymatic assays as described previously (12). Plasma LDL-C concentrations were calculated using the Friedewald formula.
Analysis of HDL size and subclass distribution
HDL size and subclass distributions were measured using two independent methods, gradient gel electrophoresis and nuclear magnetic resonance.
Gradient gel electrophoresis (GGE). Aliquots of plasma (drawn before heparin injection) were stored at -80°C and shipped on dry ice to Dr. Rainwater's laboratory. Lipoproteins in plasma were subjected to electrophoresis in nondenaturing composite gradient gels as described previously (24). After electrophoresis, lipoprotein cholesterol was stained using Sudan black B, and absorbance profiles were determined with an LKB-Ultroscan XL laser densitometer. HDL particle sizes were calibrated using a standard curve that included albumin, lactate debydrogenase, ferritin, and thyroglobulin (Pharmacia). Estimates of median HDL diameter were obtained from the absorbance profiles by determining the particle diameter at which half of the absorbance comes from larger and half from smaller HDL particles. The relative cholesterol content of the HDL subclasses was determined by dividing the lipoprotein absorbance profile into five size intervals: HDL3C, 7.2-7.8 nm; HDL3b, 7.8-8.2 nm; HDL3a, 8.2-8.8 nm; HDL2a, 8.8-9.7 nm; and HDL2b, 9.7-12.9 nm. Each sample was repeated at least twice on different gels and the average values were analyzed.
Nuclear7nagnetic resonance (NMR). Aliquots of plasma drawn before heparin injection were stored at -80°C and shipped on dry ice to Dr. Otvos' laboratory. Lipoprotein subclass profiles were measured by proton NMR spectroscopy as described previously (25, 26). This method exploits the fact that each lipoprotein particle in plasma broadcasts a distinct lipid NMR signal, the intensity of which is proportional to its lipid mass concentration. By acquiring NMR spectra of each plasma specimen (0.7 ml) on a dedicated 360 MHz spectrometer (Analogic Corp.) under defined conditions (47° C), and then deconvoluting the composite lipid methyl group signal envelope that appears in the spectrum at ~0.8 ppm, the concentrations of 15 subclasses of VLDL, LDL, and HDL are derived simultaneously The diameter ranges of the five HDL subclasses quantified by NMR were determined by calibration using purified HDL subclasses characterized by GGE. The names given to the NMR-derived HDL subclasses and their average diameters are as follows: H1, 7.5 ± 0.2 nm; H2, 8.0 ± 0.2 nm; H3, 8.5 ± 0.3 nm; H4, 9.4 ± 0.6 nm; H5 11.5 ± 1.5 nm. The size ranges of these subclasses match those measured by GGE, therefore they will be referred to as HDL3c, HDL3b, HDL3a, HDL2a, and HDL2b respectively. Concentrations of the HDL subclasses are expressed in units of cholesterol (mg/dL) and mean HDL particle size was determined by weighting the relative percentage of each HDL subclass by its diameter.
Statistical methods
Correlations between hepatic lipase activity, plasma triglyceride concentrations, HDL size, subclass concentrations, and relative subclass distributions were determined using Spearmans rank procedure. The mean and median values of each measured parameter in -514C and -514T homozygotes were compared using unpaired t-tests and Wilcoxon rank tests, respectively The effects of stanozolol treatment on the mean and median values of each parameter were assessed using paired l-tests and paired Wilcoxon rank tests.
RESULTS
Hepatic lipase genotype and HDL size distribution
Hepatic lipase activity was significantly lower in the 6 men who were -514T homozygotes than in the 11 men who were -514C homozygotes (Table 1). Plasma triglyceride, cholesterol, and HDL cholesterol concentrations were similar in the two groups (Table 1) . HDL sizes tended to be larger in -514T than in -514C homozygotes, but the differences did not reach statistical significance (Table 2). Both NMR and GGE indicated that the proportion of HDL-C in the largest HDL subclass (HDL2b) was significantly higher, and the proportion of HDL-C in the HDL3a subclass was significantly lower in homozygotes for the -514T allele than in homozygotes for the -514C allele (Table 2).
TABLE 1. Plasma lipid and lipoprotein concentrations in normolipidemic men with the -514C or -514T hepatic lipase genotypes Variable
-514C (n = 11)
-514T (n = 6)
Body Mass Index (kg× m-2)
26 ± 3
25 ± 2
Hepatic lipase activity (mmol× l-1× hr-1)
45 ± 15a*
20 ± 7
Total cholesterol (mg/dl)
162 ± 26
173 ± 25
Total triglyceride (mg/dl)
95 ± 36
78 ± 30
LDL-C (mg/dl)
107 ± 28
120 ± 31
HDL-C (mg/dl)
40 ± 7
43 ± 7
Plasma total cholesterol, triglyceride, LDL, and HDL concentrations were determined by standard enzymatic methods. Very similar estimates of these parameters were obtained by NMR spectroscopy (not shown).
*P < 0.01 for -514C versus -514T (Wilcoxon ranked sum test).
TABLE 2. HDL size and subclass distributions in normolipidemic men with the—514C or 514T hepatic lipase genotypes
-514C (n = 11)
-514T (n = 6)
Pvalue
NMR
HDL size (nm)
8.7 ± 0.4
9.0 ± 0.4
0.36
HDL2b (mg/dl)
4.6 ± 2.5
7.4 ± 3.2
0.04
HDL2a (mg/dl)
6.6 ± 7.8
6.3 ± 4.7
0.95
HDL3a (mg/dl)
11.0 ± 3.5
6.1 ± 2.1
0.01
HDL3b (mg/dl)
9.4 ± 5.7
8.8 ± 6.3
0.88
HDL3c (mg/dl)
7.6 ± 4.8
10.9 ± 6.1
0.26
GGE
HDL size (nm)
8.6 ± 0.3
8.9 ± 0.3
0.14
HDL2b (%)
17 ± 8
33 ± 7
0.005
HDL2a (%)
22 ± 6
20 ± 3
0.60
HDL3a (%)
32 ± 6
24 ± 3
0.007
HDL3b (%)
22 ± 8
17 ± 6
0.21
HDL3c (%)
7 ± 5
7 ± 5
0.90
P values were determined by Wilcoxon rank sum test.
Effects of stanozolol on hepatic lipase activity and HDL size distribution
Stanozolol administration markedly increased hepatic lipase activity and decreased HDL-C concentrations in all 20 men. Plasma triglyceride and LDL-C concentrations were not significantly altered by the drug (Table 3). Mean HDL size was reduced in all of the men. NMR spectroscopy revealed that the decrease in HDL size reflected a decrease in the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a) (Table 4). The plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were not affected by stanozolol treatment.
TABLE 3. Effects of stanozolol on plasma lipid and lipoprotein concentrations in 20 normolipidemic men Hepatic Lipase
Cholesterol
Triglycende
LDL-C
HDL-C
mmol× l-1× hr-1
mg/dl
Baseline
38 ± 18*
164 ± 29
117 ± 48
111 ± 29
39 ± 7
Stanozolol
87 ± 25
154 ± 44
106 ± 47
118 ± 41
23 ± 5
Values in the table are means ± standard deviations from 20 men given stanozolol (0.2 mg/kg/day to a maximum of 16 mg/day) orally for 6 days.
*P < 0.001 (paired Wilcoxon ranked sum tests).
TABLE 4. Effects of stanozolol on HDL size and subclass distributions in 20 normolipidemic men Baseline
Stanozolol
PValue
NMR
HDL size (nm)
8.8 ± 0.4
8.4 ± 0.3
0.0001
HDL2b (mg/dl)
4.9 ± 2.8
1.9 ± 1.5
0.0001
HDL2a (mg/dl)
7.1 ± 6.7
0.8 ± 1.4
0.0003
HDL3a (mg/dl)
9.5 ± 3.2
6.1 ± 3.2
0.0014
HDL3b (mg/dl)
10.4 ± 5.4
9.1 ± 5.1
0.25
HDL3c (mg/dl)
7.1 ± 4.2
5.0 ± 4.0
0.25
GGE
HDL size (nm)
8.7 ± 0.3
8.3 ± 0.2
0.0001
HDL2b (%)
19 ± 10
15 ± 5
0.14
HDL2a (%)
22 ± 7
12 ± 7
0.0001
HDL3a (%)
32 ± 7
24 ± 8
0.003
HDL3b (%)
21 ± 9
36 ± 9
0.0001
HDL3c (%)
6 ± 4
13 ± 6
0.0001
Values in the table are means ± standard deviations from 20 men given stanozolol (0.2 mg/kg/day to a maximum of 16 mg/day) orally for 6 days. P values were determined using paired Wilcoson ranked sum tests.
Correlation between HDL sizes determined by NMR and GGE
The values of the mean HDL diameters obtained using the two methods were similar in magnitude (means and standard deviations were 8.8 ± 0.4 and 8.7 ± 0.3 nm for NMR and GGE, respectively) and were highly correlated (r = 0.88). The percentages of HDL in each subclass (HDL2b, HDL2a, HDL3a, HDL3b, HDL3c) determined using NMR were 14 ± 7, 14 ± 13, 24 ± 9, 25 ± 15, and 23 ± 16, respectively. The corresponding percentages determined using GGE were 22 ± 10, 21 ± 6, 31 ± 7, 20 ± 8, and 7 ± 4, respectively.
Correlations between hepatic lipase activity, plasma lipid and lipoprotein concentrations, and HDL size distribution
Hepatic lipase activity was negatively correlated with HDL-C but the correlation coefficient observed did not reach statistical significance at the 5% confidence level (Table 5). Strong, statistically significant negative correlations were observed between hepatic lipase activity and the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a) (Table 5). Hepatic lipase activity was not significantly correlated with the concentrations of small HDL (HDL3b and HDL3c) determined by NMR but was positively correlated with the percentage of HDL in these subclasses as determined by GGE. Plasma triglyceride concentrations were significantly negatively correlated with HDL-C concentration, and with HDL size and subclass distributions (Table 5). Hepatic lipase activity was positively correlated with plasma triglyceride concentrations, but not with plasma or LDL cholesterol concentrations.
TABLE 5. Correlations between hepatic lipase activity, plasma lipid and lipoprotein concentrations, and HDL size and subclass distribution determined by NMR and GGE in 21 normolipidemic men Hepatic Lipase Activity
Plasma Triglyceride Concentration
Plasma total cholesterol
0.16
0.44
Plasma triglyceride
0.56
1
LDL-C
0.07
0.29
HDL-C
-0.33
- 0 57
NMR
HDL size
-0.64
-0.71
HDL2b
-0.64
-0.55
HDL2a
-0.41
-0.69
HDL3a
0.49
0.29
HDL3b
0.17
0.20
HDL3C
0.04
0.39
GGE
HDL sue
-0.70
-0.72
HDL2b
-0.62
-0.59
HDL2a
-0.47
-0.74
HDL3a
0.37
0.19
HDL3b
0.64
0.75
HDL3C
0.59
0.68
Correlation coefficients greater than 0.37 or less than -0.37 are statistically significant (P < 0.05).
DISCUSSION
HDL is a heterogenous group of lipoproteins that differ in size and composition (27-29). Several factors including gender, diet, and hypolipidemic drugs preferentially influence specific HDL subclasses, but the metabolic determinants of HDL subclass distribution have not been fully elucidated. In the present study, the relationship between hepatic lipase activity and HDL subclass distribution was investigated using proton NMR and GGE techniques. The study included two components. First, the subclass distribution of HDL was compared in normotriglyceridemic white men who were homozygotes for either the -514T or the -514C allele of hepatic lipase. Homozygotes for the -514T allele, who have genetically low hepatic lipase activity (13), had higher plasma concentrations of the largest HDL subclass, HDL2b, and lower plasma concentrations of a smaller HDL subclass, HDL3a, than did homozygotes for the -514C allele. These differences were independent of plasma triglyceride and total HDL concentrations, which were similar in -514T and -514C homozygotes as a consequence of the selection procedure. This finding provides direct evidence that primary variation in hepatic lipase activity influences HDL subclass distribution. Second, the synthetic anabolic steroid stanozolol was used to increase hepatic lipase activity. Stanozolol treatment markedly decreased the plasma concentrations of the largest HDL subclasses (HDL2b and HDL2a) but did not affect concentrations of the smallest HDL subclasses (HDL3b and HDL3c). Taken together, our data indicate that variation in hepatic lipase activity within the normal range of the population has highly specific effects on HDL size distribution among normotriglyceridemic men.
The results of the present study support and extend the findings of previous studies in two respects. First, by comparing normotriglyceridemic men with genetically defined differences in hepatic lipase activity, the problem of confounding by secondary factors is avoided. The mean hepatic lipase activity of the -514T homozygotes in this study corresponds to the 10th percentile for white men studied in our laboratory, whereas the mean value for the -514C homozygotes is equivalent to the 65th percentile (J. C. Cohen, unpublished results). This difference in hepatic lipase activity is due exclusively to DNA sequence differences between the -514T and -514C alleles, and is independent of secondary factors such as obesity or insulin resistance. Plasma triglyceride concentrations, which have a major influence on lipoprotein size distributions (30, 31), were similar in the two genotype groups. Therefore the differences in HDL size distribution between -514T and -514C homozygotes are almost certainly the direct result of genetic differences in their hepatic lipase activities.
Second, most previous studies have examined the relationship between hepatic lipase and the two major HDL subclasses, HDL2and HDL3 (4, 6, 32). The methods used in the present study allowed further resolution of HDL into five subclasses, and indicated that primary variation in hepatic lipase activity within the normal range of the population is associated with highly specific differences in HDL subclass distribution. Homozygotes for the -514T allele had higher plasma concentrations of HDL2b and lower concentrations of HDL3a compared with -514C homozygotes. Highly consistent results were obtained using two independent methods (NMR and GGE) therefore it is extremely unlikely that this result is an artifact of the analytical procedure. Accordingly, this finding indicates that low hepatic lipase activity leads to high plasma concentrations of the HDL2b subclass, and a relative decrease in the concentration of the HDL3a subclass. The plasma concentrations of the other three HDL subclasses (HDL2a, HDL3b, and HDL3c) do not appear to be significantly affected by a low hepatic lipase activity.
To date no hepatic lipase allele associated with markedly elevated hepatic lipase activity has been identified. Accordingly we analyzed the effects of pharmacological stimulation of hepatic lipase activity on HDL subclass distribution using the synthetic anabolic steroid, stanozolol. After 6 days of stanozolol treatment, mean hepatic lipase activity increased more than 2-fold, reaching the upper end of the normal range (>95th percentile) for white men measured in our laboratory. Mean plasma HDL-C decreased by more than 40%. NMR spectroscopy indicated that the major decrease occurred in the larger HDL subclasses (HDL2b and HDL2a) with a smaller decrease in HDL3a and no change in the plasma concentrations of the smallest HDL subfractions, HDL3b and HDL3c. This resulted in a decrease in the proportion of lipid staining in the larger HDL, and an increase in the proportion of lipid staining in smaller HDL, as indicated by GGE. The observation that the plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were not affected by stanozolol treatment despite a 2-fold increase in hepatic lipase activity clearly indicates that variation in hepatic lipase activity does not affect these lipoprotein subclasses. This finding is consistent with the observation that plasma concentrations of HDL3b and HDL3c were similar in men with genetically defined differences in hepatic lipase activity. The decreased concentrations of the larger HDL subclasses during stanozolol treatment are consistent with the notion that high hepatic lipase activity leads to a decrease in the plasma concentrations of large HDL. However, a recent study in two hepatic lipase-deficient patients indicates that the effect of stanozolol on plasma HDL concentrations and subclass distribution cannot be fully accounted for by changes in hepatic lipase activity (33). In that study, Bausserman, Saritelli, and Herbert (33) found that 7 days of stanozolol treatment (a duration similar to that used in the present study) led to a decrease in HDL2 concentrations in one patient but not in the other. After 14 days of stanozolol treatment, plasma concentrations of HDL2 decreased by 40% in one patient and by 59% in the other, compared with a 90% decrease in normal control subjects. Although the plasma lipid and lipoprotein concentrations of these two hepatic lipase-deficient patients differed considerably from those of the men in the present study (both patients were hypertriglyceridemic and both had a marked preponderance of HDL2 in their HDL fraction), these findings suggest that the effects of stanozolol on the HDL2b and HDL2a subclasses observed in the present study may not be entirely due to increased hepatic lipase activity.
The HDL2b subclass identified by GGE is equivalent to the HDL2 subclass isolated by zonal ultracentrifugation (34, 35). This subclass is enriched in all lipid constituents (free and unesterified cholesterol, phospholipids, and triglycerides) and has a lower protein content relative to HDL3 (35, 36). Interestingly, the apolipoprotein A-II content of HDL2 is markedly reduced relative to HDL3 (36). The results of the present study, therefore, suggest that large, lipid-enriched HDL that are relatively deficient in apolipoprotein A-II accumulate in men with low hepatic lipase activity, and are preferentially catabolized in men with high hepatic lipase activity.
In summary, the results of the present study provide direct evidence that normal variation in hepatic lipase activity is associated with specific differences in the subclass distribution of HDL among normolipidemic men. Low hepatic lipase activity leads to a specific increase in the largest HDL subclass, HDL2b. Our data suggest that high hepatic lipase activity leads to low plasma concentrations of the HDL2b and HDL2a subclasses, although direct evidence for this association must await the identification of a hepatic lipase allele that confers genetically high hepatic lipase activity. Finally, our data clearly indicate that wide variation in hepatic lipase activity has little effect on the plasma concentrations of the smallest HDL particles, HDL3b and HDL3c.
We thank Sharon Haynes R. N., Perry Moore, Jr., Hanh Nguyen, Biman Pramanik, Wendy Shelledy, and Han Tron for excellent technical assistance, Dick Verstraete for subject recruitment, and Ann White for helpful discussion. This work was supported by National Institutes of Health grants HL-53917 (J. C. C.), HL-45522 (D. L. R) and M-01-RR-00633 (S. M. G.), the Department of Veterans Affairs, the Southwestern Medical Foundation, and the Moss Heart Foundation, Dallas.
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16. Shirai, K, R. L. Barnhart, and R. L.Jackson. 1981. Hydrolysis of human plasma high density lipoprotein 2- phospholipids and triglycerides by hepatic lipase. Biochem. Biophys. Res. Commun. 100: 591-599.
17. Davis, C. E., D. Gordon,J. LaRosa, R D. Wood, and M. Halperin. 1980. Correlations of plasma high-density lipoprotein cholesterol levels with other plasma lipid and lipoprotein concentrations. Circulation. 62: IV24-lV30.
18. Williams, R. D. Robinson, and A. Bailey. 1979. High-density lipoprotein and coronary risk factors in normal men. Lancet. 1: 7275.
19. Schaefer, E. J., R. I. Levy, D. W. Anderson, R. N. Danner, H. B. Brewer, Jr., and W. C. Blackwelder. 1978. Plasma-triglycerides in regulation of HDL-cholesterol levels. Lancet. 2: 391-393.
20. Albrink, M. J., R. M. Krauss, F. T. Lindgrem,J. von der Groeben, S. Pan, and R D. Wood. 1980. Intercorrelations among plasma high density lipoproteins, obesity and triglycerides in a normal population. Lipids. 15: 668-676.
21. Knudsen, R. M. Antikainen, S. Ehnholm, M. Uusi-Oukari, H. Tenkanen, S. Lahdenpera,J. Kahri, M. Tilly-Kiesi, A. Bensadoun, M. R. Taskinen, and C. Ehnholm. 1996. A compound heterozygote for hepatic lipase gene mutations Leu334~Phe and Thr383~Met: correlahon between hepahc lipase activity and phenotypic expression. J. Lipid Res. 37: 825-834.
22. Baginsky, M. L., and W. V. Brown. 1979. A new method for the measurement of lipoprotein lipase in postheparin plasma using sodium dodecyl sulfate for the inactivation of hepatic triglyceride lipase. J. Lipid Res. 20: 548-556.
23. Belfrage, R, and M. Vaughan. 1969. Simple liquid-liquid partition system for isolation of labeled oleic acid from mixtures with glycerides. J. Lipid Res. 10: 341-344.
24. Rainwater, D. L., R H. J. Moore, W. R. Shelledy, T. D. Dyer, and S. H. Slifer. 1997. Characterization of a composite gradient gel for the electrophoretic separation of lipoproteins. J. Lipid Res. 38: 1261-1266.
25. Otvos,J. D., E.J.Jeyarajah, D. W. Bennett, and R. M. Krauss. 1992. Development of a proton nuclear magnetic resonance spectroscopic method for determining plasma lipoprotein concentrations and subspecies distributions from a single, rapid measurement. Clin. Chem. 38: 1632-1638.
26. Otvos,J. D. 1997. Measurement of lipoprotein subclass profiles by nuclear magnetic resonance spectroscopy. In Handbook of Lipoprotein Testing. Anonymous, AACC Press, Washington, DC. 497508.
27. Skinner, E. R. 1994. High-density lipoprotein subclasses. Curr Opin. Lipidol. 5: 241-247.
28. von, E. A., Y. Huang, and G. Assmann. 1994. Physiological role and clinical relevance of high density lipoprotein subclasses. Curr Opin. Lipidol. 5:404 416.
29. Tailleux, A., and J. C. Fruchart. 1996. HDL heterogeneity and atherosclerosis. Crit. Rev. Clin. Lab. Sa. 33: 163-201.
30. McNamara, J. R., J. L. Jenner, Z. Li, R W. Wilson, and E. J. Schaefer. 1992. Change in LDL particle size is associated with change in plasma triglyceride concentration. Arterioscler. Thromb. 12: 1284 1290.
31. Johansson,J., G. Walldius, and L. A. Carlson. 1992. Close correlation between high-density lipoprotein and triglycerides in normotriglyceridaemia. J. Intern. Med. 232: 43-51.
32. Patsch, J. R., S. Prasad, A. M. J. Gotto, Jr., and W. Patsch. 1987. High density lipoprotein2. Relationship of the plasma levels of this lipoprotein species to its composition, to the magnitude of postprandial lipemia, and to the activities of lipoprotein lipase and hepatic lipase. J. Clin. Invest. 80: 341-347.
33. Bausserman, L. L., A. L. Saritelli, and R N. Herbert. 1997. Effects of short-term stanozolol administration on serum lipoproteins in hepatic lipase deficiency Metabolism. 46: 992-996.
34. Patsch,J. R., G. Kostner, and W. Patsch. 1983. Separation and analysis of plasma lipoproteins by zonal ultracentrifugation. In CRC Handbook of Electrophoresis. L. Lewis and J. Opplt, editors. CRC Press Iric, Boca Raton, FL. 67-81.
35. McNerney, C. A., M. L. Kashyap, R. L. Barnhart, and R. L.Jackson. 1985. Comparison of gradient gel electrophoresis and zonal ultracentrifugation for quantitation of high density lipoproteins. J. Lipid Res. 26: 1363-1367.
36. Patsch, W., G. Schonfeld, A.M. Gotto,Jr., andJ. R. Patsch. 1980. Characterization of human high density lipoproteins by zonal ultracentrifugation. J. Biol. Chem. 255: 3178-3185.
you would be better off stacking with test, even used with arimidex you will get some estrogen (which will not happen with winstrol)
NOTE- STACKING WITH DECA MAY EXACERBATE THE HDL PROBLEM... AND IT DECREASES THE BENEFICIAL EFFECTS OF ANY SERM OR NATURAL ESTROGEN PRESENT...
Hepatic lipase activity influences high density lipoprotein subclass distribution in normotriglyceridemic men: genetic and pharmacological evidence
Scott M. Grundy, Gloria L. Vega, James D. Otvos,
David L. Rainwater, and Jonathan C. Cohen
Abbreviations:
HDL-C, high density lipoprotein cholesterol;
GGE, gradient gel electrophoresis;
NMR, nuclear magnetic resonance.
Abstract: Several studies have reported an inverse relationship between hepatic lipase activity and plasma high density lipoprotein (HDL) cholesterol concentrations. The purpose of the present study was to determine whether genetic and pharmacological variation in hepatic lipase activity alters the distribution of HDL subclasses. Two independent analytical methods (nuclear magnetic resonance and gradient gel electrophoresis) were used to compare HDL subclass distributions in 11 homozygotes for the -514C allele of hepatic lipase and in 6 homozygotes for the -514T allele. Mean hepatic lipase activity was 45 ± 15 mmol× l-1× hr-1 in -514C homozygotes and 20 ± 7 mmol× l-1× hr-1 in -514T homozygotes. Both analytical methods indicated that HDL2b was significantly higher and HDL3a was significantly lower in -514T homozygotes than in -514C homozygotes. No differences were noted in the other HDL fractions (HDL2a, HDL3b, and HDL3c). To determine the effects of increased hepatic lipase activity, 20 men were given the synthetic anabolic steroid, stanozolol. Stanozolol treatment increased hepatic lipase activity more than two-fold (38 ± 18 to 85 ± 25 mmol× l-1× hr-1), and markedly reduced the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a). The plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were unchanged by stanozolol treatment. Taken together, these genetic and pharmacological data indicate that variation in hepatic lipase activity has highly specific effects on the distribution of HDL subclasses in the circulation. -Grundy, S. M., G. L. Vega, J. D. Otvos, D. L. Rainwater, and J. C. Cohen. Hepatic lipase activity influences high density lipoprotein subclass distribution in normotriglyceridemic men: genetic and pharmacological evidence. J. Lipid Res. 1999. 40: 229-234.
Human hepatic lipase is a 476 amino acid glycoprotein that exhibits phospholipase A1 and triglyceride hydrolase activities (1, 2). The enzyme is synthesized in hepatocytes, secreted, and bound to hepatocyte and hepatic endothelial surfaces (3), from which it can be released into the circulation by heparin. Data from several clinical, pharmacological, and genetic studies support an inverse relationship between hepatic lipase activity and the plasma concentrations of high density lipoprotein cholesterol (HDL-C) in humans. The activity of hepatic lipase in postheparin plasma appears to be inversely correlated with plasma HDL-C concentrations among unselected individuals (4-8). Drugs such as anabolic steroids that increase hepatic lipase activity decrease HDL concentrations (9), whereas hepatic lipase deficiency is associated with high plasma HDL levels in some patients (10). Studies of normolipidemic families indicated linkage between plasma HDL concentrations and LIPC, the gene encoding hepatic lipase (11, 12). DNA sequencing revealed an LIPC allele (designated -514T) containing four linked polymorphisms in the 5' flanking region. This allele was associated with low hepatic lipase activity (13) and increased plasma HDL-C concentrations in white men (12). These findings suggest that hepatic lipase plays a role in the catabolism of HDL, and that normal variation in hepatic lipase activity is associated with inter-individual differences in plasma HDL concentration.
Some studies have indicated that hepatic lipase activity is correlated with the plasma concentrations of specific HDL subfractions. Kuusi, Saarinen, and Nikkila (4) reported that hepatic lipase activity was inversely correlated with plasma concentrations of HDL2, but not HDL3 determined by ultracentrifugation. Katzel et al. (14) found that postheparin plasma hepatic lipase activity was inversely correlated with the proportion of HDL in the HDL2b subspecies determined by gradient gel electrophoresis (GGE) in 41 healthy men.Johansson et al. (15) reported that hepatic lipase was inversely correlated with HDL2b and positively correlated with HDL3b (determined by GGE) in 24 normolipidemic infarct survivors. These observations are consistent with in vitro studies indicating that hepatic lipase acts preferentially on large HDL particles (16), and suggest that hepatic lipase plays a role in the remodeling of circulating HDL. However, all of the clinical studies performed to date were based on correlations observed between two phenotypes, hepatic lipase activity and HDL subclass distribution. Such correlations are subject to confounding by secondary factors and do not constitute definitive evidence for a causal relationship between hepatic lipase activity and plasma HDL-C concentrations. For example, plasma triglyceride concentrations are positively correlated with hepatic lipase activity (5) and inversely related to plasma HDL-C concentrations ( 17-20) . Therefore a correlation between hepatic lipase activity and plasma HDL size among unselected individuals may simply reflect the interactions of plasma triglyceride concentrations (or some other confounder) with both parameters.
Secondary confounding can be avoided by studying individuals with genetic (i.e., primary) variation in hepatic lipase activity. Detailed analysis of HDL subclass distribution in such subjects is limited to a single report by Knudsen et al. (20), who studied a compound heterozygote for two hepatic lipase gene mutations. Hepatic lipase activity in this individual was decreased by 70% compared to healthy control men. Density gradient ultracentrifugation of his HDL revealed markedly increased concentrations of the IDLE and HDL2b subclasses. The present study was undertaken to determine whether variation in hepatic lipase activity alters the distribution of HDL subclasses. To minimize confounding by secondary factors, HDL subclass distributions were measured in normotriglyceridemic men with genetically defined differences in hepatic lipase activity. To ensure reliable estimation of HDL subclass distribution, two independent analytical methods (nuclear magnetic resonance and gradient gel electrophoresis) were used. To support the findings of this genetic approach, the effects of pharmacological stimulation of hepatic lipase activity on HDL size distribution was determined.
METHODS
Experimental design
Fasting blood samples were drawn from 21 normolipidemic white men. Six of the men were homozygous for the -514T allele of LIPC, four were -514C/ -514T heterozygotes, and 11 were -514C homozygotes. Five of the -514T homozygotes and five of the -514C homozygotes had been matched for plasma HDL-C concentrations in a previous study Aliquots of plasma were stored at 4°C for lipid and lipoprotein measurement, and at -80°C for HDL subclass analysis. Heparin injections for determination of postheparin plasma hepatic lipase activity were performed after an overnight fast. Twenty of the men then took 0.2 mg/kg per day of the oral synthetic anabolic steroid stanozolol (Winstrol, Winthrop, Pittsburg, PA) up to a maximum of 16 ma/ day for 6 consecutive days. Compliance was monitored by pill counting. The drug was well tolerated by all of the men, and no side effects were reported. A second heparin injection was performed within 24 h after ingestion of the final dose of stanozolol.
Subjects
The study was approved by the Institutional Review Board at the University of Texas Southwestern Medical Center. Men aged 20 to 50 years who were apparently healthy, who had plasma triglyceride concentrations below 200 mg/dL, and who did not smoke or use lipid-lowering drugs or beta blockers were recruited by advertisements placed on notice boards at Southwestern Medical Center. Subjects were studied as outpatients on their ad libitum diets.
Hepatic lipase genotyping
Hepatic lipase genotypes were determined by PCR amplification and restriction enzyme digestion as described in detail previously (12).
Assay of hepatic lipase activity
Hepatic lipase activity was measured in postheparin plasma using a modification of the method reported by Baginsky and Brown (22) as described in detail previously (8). Briefly, patients received an intravenous injection of 75 IU of porcine heparin (Elkins-Sinn Inc.) per kg of body weight after a 12-h fast. Blood samples were drawn into tubes containing EDTA before and 15 min after injection. Plasma was separated immediately upon collection of blood and aliquots were frozen at -70°C. Hepatic lipase activity was measured by addition of 0.025 mL postheparin plasma to 0.475 mL medium (pH 8.8) containing NaCI (0.75 m), a gum arable-stabilized emulsion of triolein (final concentration 15 mmol/L) labeled with [3H]triolein (0.5 m Ci), and fatty acid-free bovine serum albumin (final concentration 50 g/L). The free [3H]oleate generated during the hydrolysis reactions was extracted using the method of Belfrage and Vaughan (23). The intra- and inter-assay coefficients of variation were <5%.
Assay of plasma lipids and lipoproteins
Plasma triglyceride, cholesterol, and HDL-C concentrations were measured using standard enzymatic assays as described previously (12). Plasma LDL-C concentrations were calculated using the Friedewald formula.
Analysis of HDL size and subclass distribution
HDL size and subclass distributions were measured using two independent methods, gradient gel electrophoresis and nuclear magnetic resonance.
Gradient gel electrophoresis (GGE). Aliquots of plasma (drawn before heparin injection) were stored at -80°C and shipped on dry ice to Dr. Rainwater's laboratory. Lipoproteins in plasma were subjected to electrophoresis in nondenaturing composite gradient gels as described previously (24). After electrophoresis, lipoprotein cholesterol was stained using Sudan black B, and absorbance profiles were determined with an LKB-Ultroscan XL laser densitometer. HDL particle sizes were calibrated using a standard curve that included albumin, lactate debydrogenase, ferritin, and thyroglobulin (Pharmacia). Estimates of median HDL diameter were obtained from the absorbance profiles by determining the particle diameter at which half of the absorbance comes from larger and half from smaller HDL particles. The relative cholesterol content of the HDL subclasses was determined by dividing the lipoprotein absorbance profile into five size intervals: HDL3C, 7.2-7.8 nm; HDL3b, 7.8-8.2 nm; HDL3a, 8.2-8.8 nm; HDL2a, 8.8-9.7 nm; and HDL2b, 9.7-12.9 nm. Each sample was repeated at least twice on different gels and the average values were analyzed.
Nuclear7nagnetic resonance (NMR). Aliquots of plasma drawn before heparin injection were stored at -80°C and shipped on dry ice to Dr. Otvos' laboratory. Lipoprotein subclass profiles were measured by proton NMR spectroscopy as described previously (25, 26). This method exploits the fact that each lipoprotein particle in plasma broadcasts a distinct lipid NMR signal, the intensity of which is proportional to its lipid mass concentration. By acquiring NMR spectra of each plasma specimen (0.7 ml) on a dedicated 360 MHz spectrometer (Analogic Corp.) under defined conditions (47° C), and then deconvoluting the composite lipid methyl group signal envelope that appears in the spectrum at ~0.8 ppm, the concentrations of 15 subclasses of VLDL, LDL, and HDL are derived simultaneously The diameter ranges of the five HDL subclasses quantified by NMR were determined by calibration using purified HDL subclasses characterized by GGE. The names given to the NMR-derived HDL subclasses and their average diameters are as follows: H1, 7.5 ± 0.2 nm; H2, 8.0 ± 0.2 nm; H3, 8.5 ± 0.3 nm; H4, 9.4 ± 0.6 nm; H5 11.5 ± 1.5 nm. The size ranges of these subclasses match those measured by GGE, therefore they will be referred to as HDL3c, HDL3b, HDL3a, HDL2a, and HDL2b respectively. Concentrations of the HDL subclasses are expressed in units of cholesterol (mg/dL) and mean HDL particle size was determined by weighting the relative percentage of each HDL subclass by its diameter.
Statistical methods
Correlations between hepatic lipase activity, plasma triglyceride concentrations, HDL size, subclass concentrations, and relative subclass distributions were determined using Spearmans rank procedure. The mean and median values of each measured parameter in -514C and -514T homozygotes were compared using unpaired t-tests and Wilcoxon rank tests, respectively The effects of stanozolol treatment on the mean and median values of each parameter were assessed using paired l-tests and paired Wilcoxon rank tests.
RESULTS
Hepatic lipase genotype and HDL size distribution
Hepatic lipase activity was significantly lower in the 6 men who were -514T homozygotes than in the 11 men who were -514C homozygotes (Table 1). Plasma triglyceride, cholesterol, and HDL cholesterol concentrations were similar in the two groups (Table 1) . HDL sizes tended to be larger in -514T than in -514C homozygotes, but the differences did not reach statistical significance (Table 2). Both NMR and GGE indicated that the proportion of HDL-C in the largest HDL subclass (HDL2b) was significantly higher, and the proportion of HDL-C in the HDL3a subclass was significantly lower in homozygotes for the -514T allele than in homozygotes for the -514C allele (Table 2).
TABLE 1. Plasma lipid and lipoprotein concentrations in normolipidemic men with the -514C or -514T hepatic lipase genotypes Variable
-514C (n = 11)
-514T (n = 6)
Body Mass Index (kg× m-2)
26 ± 3
25 ± 2
Hepatic lipase activity (mmol× l-1× hr-1)
45 ± 15a*
20 ± 7
Total cholesterol (mg/dl)
162 ± 26
173 ± 25
Total triglyceride (mg/dl)
95 ± 36
78 ± 30
LDL-C (mg/dl)
107 ± 28
120 ± 31
HDL-C (mg/dl)
40 ± 7
43 ± 7
Plasma total cholesterol, triglyceride, LDL, and HDL concentrations were determined by standard enzymatic methods. Very similar estimates of these parameters were obtained by NMR spectroscopy (not shown).
*P < 0.01 for -514C versus -514T (Wilcoxon ranked sum test).
TABLE 2. HDL size and subclass distributions in normolipidemic men with the—514C or 514T hepatic lipase genotypes
-514C (n = 11)
-514T (n = 6)
Pvalue
NMR
HDL size (nm)
8.7 ± 0.4
9.0 ± 0.4
0.36
HDL2b (mg/dl)
4.6 ± 2.5
7.4 ± 3.2
0.04
HDL2a (mg/dl)
6.6 ± 7.8
6.3 ± 4.7
0.95
HDL3a (mg/dl)
11.0 ± 3.5
6.1 ± 2.1
0.01
HDL3b (mg/dl)
9.4 ± 5.7
8.8 ± 6.3
0.88
HDL3c (mg/dl)
7.6 ± 4.8
10.9 ± 6.1
0.26
GGE
HDL size (nm)
8.6 ± 0.3
8.9 ± 0.3
0.14
HDL2b (%)
17 ± 8
33 ± 7
0.005
HDL2a (%)
22 ± 6
20 ± 3
0.60
HDL3a (%)
32 ± 6
24 ± 3
0.007
HDL3b (%)
22 ± 8
17 ± 6
0.21
HDL3c (%)
7 ± 5
7 ± 5
0.90
P values were determined by Wilcoxon rank sum test.
Effects of stanozolol on hepatic lipase activity and HDL size distribution
Stanozolol administration markedly increased hepatic lipase activity and decreased HDL-C concentrations in all 20 men. Plasma triglyceride and LDL-C concentrations were not significantly altered by the drug (Table 3). Mean HDL size was reduced in all of the men. NMR spectroscopy revealed that the decrease in HDL size reflected a decrease in the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a) (Table 4). The plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were not affected by stanozolol treatment.
TABLE 3. Effects of stanozolol on plasma lipid and lipoprotein concentrations in 20 normolipidemic men Hepatic Lipase
Cholesterol
Triglycende
LDL-C
HDL-C
mmol× l-1× hr-1
mg/dl
Baseline
38 ± 18*
164 ± 29
117 ± 48
111 ± 29
39 ± 7
Stanozolol
87 ± 25
154 ± 44
106 ± 47
118 ± 41
23 ± 5
Values in the table are means ± standard deviations from 20 men given stanozolol (0.2 mg/kg/day to a maximum of 16 mg/day) orally for 6 days.
*P < 0.001 (paired Wilcoxon ranked sum tests).
TABLE 4. Effects of stanozolol on HDL size and subclass distributions in 20 normolipidemic men Baseline
Stanozolol
PValue
NMR
HDL size (nm)
8.8 ± 0.4
8.4 ± 0.3
0.0001
HDL2b (mg/dl)
4.9 ± 2.8
1.9 ± 1.5
0.0001
HDL2a (mg/dl)
7.1 ± 6.7
0.8 ± 1.4
0.0003
HDL3a (mg/dl)
9.5 ± 3.2
6.1 ± 3.2
0.0014
HDL3b (mg/dl)
10.4 ± 5.4
9.1 ± 5.1
0.25
HDL3c (mg/dl)
7.1 ± 4.2
5.0 ± 4.0
0.25
GGE
HDL size (nm)
8.7 ± 0.3
8.3 ± 0.2
0.0001
HDL2b (%)
19 ± 10
15 ± 5
0.14
HDL2a (%)
22 ± 7
12 ± 7
0.0001
HDL3a (%)
32 ± 7
24 ± 8
0.003
HDL3b (%)
21 ± 9
36 ± 9
0.0001
HDL3c (%)
6 ± 4
13 ± 6
0.0001
Values in the table are means ± standard deviations from 20 men given stanozolol (0.2 mg/kg/day to a maximum of 16 mg/day) orally for 6 days. P values were determined using paired Wilcoson ranked sum tests.
Correlation between HDL sizes determined by NMR and GGE
The values of the mean HDL diameters obtained using the two methods were similar in magnitude (means and standard deviations were 8.8 ± 0.4 and 8.7 ± 0.3 nm for NMR and GGE, respectively) and were highly correlated (r = 0.88). The percentages of HDL in each subclass (HDL2b, HDL2a, HDL3a, HDL3b, HDL3c) determined using NMR were 14 ± 7, 14 ± 13, 24 ± 9, 25 ± 15, and 23 ± 16, respectively. The corresponding percentages determined using GGE were 22 ± 10, 21 ± 6, 31 ± 7, 20 ± 8, and 7 ± 4, respectively.
Correlations between hepatic lipase activity, plasma lipid and lipoprotein concentrations, and HDL size distribution
Hepatic lipase activity was negatively correlated with HDL-C but the correlation coefficient observed did not reach statistical significance at the 5% confidence level (Table 5). Strong, statistically significant negative correlations were observed between hepatic lipase activity and the plasma concentrations of the larger HDL subclasses (HDL2b and HDL2a) (Table 5). Hepatic lipase activity was not significantly correlated with the concentrations of small HDL (HDL3b and HDL3c) determined by NMR but was positively correlated with the percentage of HDL in these subclasses as determined by GGE. Plasma triglyceride concentrations were significantly negatively correlated with HDL-C concentration, and with HDL size and subclass distributions (Table 5). Hepatic lipase activity was positively correlated with plasma triglyceride concentrations, but not with plasma or LDL cholesterol concentrations.
TABLE 5. Correlations between hepatic lipase activity, plasma lipid and lipoprotein concentrations, and HDL size and subclass distribution determined by NMR and GGE in 21 normolipidemic men Hepatic Lipase Activity
Plasma Triglyceride Concentration
Plasma total cholesterol
0.16
0.44
Plasma triglyceride
0.56
1
LDL-C
0.07
0.29
HDL-C
-0.33
- 0 57
NMR
HDL size
-0.64
-0.71
HDL2b
-0.64
-0.55
HDL2a
-0.41
-0.69
HDL3a
0.49
0.29
HDL3b
0.17
0.20
HDL3C
0.04
0.39
GGE
HDL sue
-0.70
-0.72
HDL2b
-0.62
-0.59
HDL2a
-0.47
-0.74
HDL3a
0.37
0.19
HDL3b
0.64
0.75
HDL3C
0.59
0.68
Correlation coefficients greater than 0.37 or less than -0.37 are statistically significant (P < 0.05).
DISCUSSION
HDL is a heterogenous group of lipoproteins that differ in size and composition (27-29). Several factors including gender, diet, and hypolipidemic drugs preferentially influence specific HDL subclasses, but the metabolic determinants of HDL subclass distribution have not been fully elucidated. In the present study, the relationship between hepatic lipase activity and HDL subclass distribution was investigated using proton NMR and GGE techniques. The study included two components. First, the subclass distribution of HDL was compared in normotriglyceridemic white men who were homozygotes for either the -514T or the -514C allele of hepatic lipase. Homozygotes for the -514T allele, who have genetically low hepatic lipase activity (13), had higher plasma concentrations of the largest HDL subclass, HDL2b, and lower plasma concentrations of a smaller HDL subclass, HDL3a, than did homozygotes for the -514C allele. These differences were independent of plasma triglyceride and total HDL concentrations, which were similar in -514T and -514C homozygotes as a consequence of the selection procedure. This finding provides direct evidence that primary variation in hepatic lipase activity influences HDL subclass distribution. Second, the synthetic anabolic steroid stanozolol was used to increase hepatic lipase activity. Stanozolol treatment markedly decreased the plasma concentrations of the largest HDL subclasses (HDL2b and HDL2a) but did not affect concentrations of the smallest HDL subclasses (HDL3b and HDL3c). Taken together, our data indicate that variation in hepatic lipase activity within the normal range of the population has highly specific effects on HDL size distribution among normotriglyceridemic men.
The results of the present study support and extend the findings of previous studies in two respects. First, by comparing normotriglyceridemic men with genetically defined differences in hepatic lipase activity, the problem of confounding by secondary factors is avoided. The mean hepatic lipase activity of the -514T homozygotes in this study corresponds to the 10th percentile for white men studied in our laboratory, whereas the mean value for the -514C homozygotes is equivalent to the 65th percentile (J. C. Cohen, unpublished results). This difference in hepatic lipase activity is due exclusively to DNA sequence differences between the -514T and -514C alleles, and is independent of secondary factors such as obesity or insulin resistance. Plasma triglyceride concentrations, which have a major influence on lipoprotein size distributions (30, 31), were similar in the two genotype groups. Therefore the differences in HDL size distribution between -514T and -514C homozygotes are almost certainly the direct result of genetic differences in their hepatic lipase activities.
Second, most previous studies have examined the relationship between hepatic lipase and the two major HDL subclasses, HDL2and HDL3 (4, 6, 32). The methods used in the present study allowed further resolution of HDL into five subclasses, and indicated that primary variation in hepatic lipase activity within the normal range of the population is associated with highly specific differences in HDL subclass distribution. Homozygotes for the -514T allele had higher plasma concentrations of HDL2b and lower concentrations of HDL3a compared with -514C homozygotes. Highly consistent results were obtained using two independent methods (NMR and GGE) therefore it is extremely unlikely that this result is an artifact of the analytical procedure. Accordingly, this finding indicates that low hepatic lipase activity leads to high plasma concentrations of the HDL2b subclass, and a relative decrease in the concentration of the HDL3a subclass. The plasma concentrations of the other three HDL subclasses (HDL2a, HDL3b, and HDL3c) do not appear to be significantly affected by a low hepatic lipase activity.
To date no hepatic lipase allele associated with markedly elevated hepatic lipase activity has been identified. Accordingly we analyzed the effects of pharmacological stimulation of hepatic lipase activity on HDL subclass distribution using the synthetic anabolic steroid, stanozolol. After 6 days of stanozolol treatment, mean hepatic lipase activity increased more than 2-fold, reaching the upper end of the normal range (>95th percentile) for white men measured in our laboratory. Mean plasma HDL-C decreased by more than 40%. NMR spectroscopy indicated that the major decrease occurred in the larger HDL subclasses (HDL2b and HDL2a) with a smaller decrease in HDL3a and no change in the plasma concentrations of the smallest HDL subfractions, HDL3b and HDL3c. This resulted in a decrease in the proportion of lipid staining in the larger HDL, and an increase in the proportion of lipid staining in smaller HDL, as indicated by GGE. The observation that the plasma concentrations of the smallest HDL subclasses (HDL3b and HDL3c) were not affected by stanozolol treatment despite a 2-fold increase in hepatic lipase activity clearly indicates that variation in hepatic lipase activity does not affect these lipoprotein subclasses. This finding is consistent with the observation that plasma concentrations of HDL3b and HDL3c were similar in men with genetically defined differences in hepatic lipase activity. The decreased concentrations of the larger HDL subclasses during stanozolol treatment are consistent with the notion that high hepatic lipase activity leads to a decrease in the plasma concentrations of large HDL. However, a recent study in two hepatic lipase-deficient patients indicates that the effect of stanozolol on plasma HDL concentrations and subclass distribution cannot be fully accounted for by changes in hepatic lipase activity (33). In that study, Bausserman, Saritelli, and Herbert (33) found that 7 days of stanozolol treatment (a duration similar to that used in the present study) led to a decrease in HDL2 concentrations in one patient but not in the other. After 14 days of stanozolol treatment, plasma concentrations of HDL2 decreased by 40% in one patient and by 59% in the other, compared with a 90% decrease in normal control subjects. Although the plasma lipid and lipoprotein concentrations of these two hepatic lipase-deficient patients differed considerably from those of the men in the present study (both patients were hypertriglyceridemic and both had a marked preponderance of HDL2 in their HDL fraction), these findings suggest that the effects of stanozolol on the HDL2b and HDL2a subclasses observed in the present study may not be entirely due to increased hepatic lipase activity.
The HDL2b subclass identified by GGE is equivalent to the HDL2 subclass isolated by zonal ultracentrifugation (34, 35). This subclass is enriched in all lipid constituents (free and unesterified cholesterol, phospholipids, and triglycerides) and has a lower protein content relative to HDL3 (35, 36). Interestingly, the apolipoprotein A-II content of HDL2 is markedly reduced relative to HDL3 (36). The results of the present study, therefore, suggest that large, lipid-enriched HDL that are relatively deficient in apolipoprotein A-II accumulate in men with low hepatic lipase activity, and are preferentially catabolized in men with high hepatic lipase activity.
In summary, the results of the present study provide direct evidence that normal variation in hepatic lipase activity is associated with specific differences in the subclass distribution of HDL among normolipidemic men. Low hepatic lipase activity leads to a specific increase in the largest HDL subclass, HDL2b. Our data suggest that high hepatic lipase activity leads to low plasma concentrations of the HDL2b and HDL2a subclasses, although direct evidence for this association must await the identification of a hepatic lipase allele that confers genetically high hepatic lipase activity. Finally, our data clearly indicate that wide variation in hepatic lipase activity has little effect on the plasma concentrations of the smallest HDL particles, HDL3b and HDL3c.
We thank Sharon Haynes R. N., Perry Moore, Jr., Hanh Nguyen, Biman Pramanik, Wendy Shelledy, and Han Tron for excellent technical assistance, Dick Verstraete for subject recruitment, and Ann White for helpful discussion. This work was supported by National Institutes of Health grants HL-53917 (J. C. C.), HL-45522 (D. L. R) and M-01-RR-00633 (S. M. G.), the Department of Veterans Affairs, the Southwestern Medical Foundation, and the Moss Heart Foundation, Dallas.
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HansNZ
New member
I take flax seed oil to help with HDL. Does it protect the heart by raising HDL or does it simply protect the heart in the same way that HDL does?
The reason I ask is that my HDL tests come back very low despite taking flax seed oil. If flax seed oil works its wonders by raising HDL then it isn't helping me. If it protects the heart despite low HDL then I am OK.
The reason I ask is that my HDL tests come back very low despite taking flax seed oil. If flax seed oil works its wonders by raising HDL then it isn't helping me. If it protects the heart despite low HDL then I am OK.
2 questions...
1) - Bill - can you tell me if there is a UK importer for Anabolics 2002? If so, who is it? I've been struggling to find a copy.
2) I understand from Anabolics 2000 that just the tablets are c17-aa. Is this right? If so, then surely drinking Winny will not work - but everyone says it does!
Thanks!
1) - Bill - can you tell me if there is a UK importer for Anabolics 2002? If so, who is it? I've been struggling to find a copy.
2) I understand from Anabolics 2000 that just the tablets are c17-aa. Is this right? If so, then surely drinking Winny will not work - but everyone says it does!
Thanks!
macrophage69alpha
New member
Demon Dan said:
if this STATMENT is true.. then whoever wrote that passage in the book.. HAS NO PLACE WRITING A BOOK ABOUT STEROIDS.
stanozolol is 17aa.. NO MATTER WHAT FORM LIQUID, POWDER OR TABLET
Well the book is a bit ambiguous so I thought I'd ask...
"Stanazolol also contains the same c17 methylation as Dianabol, an alteration used so that oral administration is possible." (Suggests they both are c17-aa.)
then later...
"With the structural (c17-aa) alteration, the tablets will also place a higher level of stress on the liver than the injectable (which avoids the "first pass"). (Suggests only the tab is c17-aa.)
Anyway - thanks for clearing it up.
"Stanazolol also contains the same c17 methylation as Dianabol, an alteration used so that oral administration is possible." (Suggests they both are c17-aa.)
then later...
"With the structural (c17-aa) alteration, the tablets will also place a higher level of stress on the liver than the injectable (which avoids the "first pass"). (Suggests only the tab is c17-aa.)
Anyway - thanks for clearing it up.
This is not suggesting that the injectable is not 17aa....it simply tell us that it is LESS toxic because it avoids the "big hit", all at once upon the liver, throught the portal vein after ingestion of the pills.
When you inject winny the negative affects upon the liver are spread out over two days.
When you inject winny the negative affects upon the liver are spread out over two days.count me out on this one
i, myself, never said injecting Winny was ok, only that it is better to inject Winny instead of drinking it.
As for the lowering of the good cholesterol...if you do a search you will find it here in the archives, since this was said here many times before.....
Doing Winny or Ox or another 17AA and getting your bloodtest done, then you will know.
Realgains said:
i, myself, never said injecting Winny was ok, only that it is better to inject Winny instead of drinking it.
As for the lowering of the good cholesterol...if you do a search you will find it here in the archives, since this was said here many times before.....
Doing Winny or Ox or another 17AA and getting your bloodtest done, then you will know.
Macro,
Shit man, the abstract would have been fine! I barely skimmed it..
To answer you:
>>>> Taking nolva with winstrol may help.. but Nolva will make YOU FAT(if you would like I will post the studies)... adding a SERM is ok, but clomid might be a better choice.
I'd like to see the ABSTRACTS or refs. Don't need a study to tell my Nolvadex DOESN'T make me fat though. And as an anti-estrogen in most tissues I think it actualy helps a little.
To your Clomid vs. Nolvadex issue, I like Nolvadex for all uses better. It is more potent as an anti-estrogen mg for mg, and doesn't have the technical drawback Clomid does of slightly desensitizing the pituitary to GnRH.
>> you would be better off stacking with test, even used with arimidex you will get some estrogen (which will not happen with winstrol)
Agreed. But if I were taking Test+Arim I'd still add Nolvadex to replace estrogen action in the liver. The whole point of using arimidex in the first place would be to diminish circulating estrogens, which is a very bad thing for cholesterol. If you are worried about cholesterol ALWAYS go with Nolvadex over an Arom. Inhibitor, or at least add it to.
>>> NOTE- STACKING WITH DECA MAY EXACERBATE THE HDL PROBLEM... AND IT DECREASES THE BENEFICIAL EFFECTS OF ANY SERM OR NATURAL ESTROGEN PRESENT...
Please explain. This makes no sense to me (as it would specifically relate to deca), and actually goes against quite a bit of first hand experience with Nolv and Deca. Nolvadex acts as an estrogen in the liver, plain and simple. Estrogenic action in this region of the body stimulates HDL synthesis. How does Deca specifically block out Nolvadex's estrogenic action here? I know androgens play opposing roles, but that is the whole issue. Steroids + Nolvadex to help the prob.. Are you really saying Deca without Nolvadex would be better for HDL than Deca + Nolvadx?
- Bill Llewellyn
Shit man, the abstract would have been fine! I barely skimmed it..
To answer you:
>>>> Taking nolva with winstrol may help.. but Nolva will make YOU FAT(if you would like I will post the studies)... adding a SERM is ok, but clomid might be a better choice.
I'd like to see the ABSTRACTS or refs. Don't need a study to tell my Nolvadex DOESN'T make me fat though. And as an anti-estrogen in most tissues I think it actualy helps a little.
To your Clomid vs. Nolvadex issue, I like Nolvadex for all uses better. It is more potent as an anti-estrogen mg for mg, and doesn't have the technical drawback Clomid does of slightly desensitizing the pituitary to GnRH.
>> you would be better off stacking with test, even used with arimidex you will get some estrogen (which will not happen with winstrol)
Agreed. But if I were taking Test+Arim I'd still add Nolvadex to replace estrogen action in the liver. The whole point of using arimidex in the first place would be to diminish circulating estrogens, which is a very bad thing for cholesterol. If you are worried about cholesterol ALWAYS go with Nolvadex over an Arom. Inhibitor, or at least add it to.
>>> NOTE- STACKING WITH DECA MAY EXACERBATE THE HDL PROBLEM... AND IT DECREASES THE BENEFICIAL EFFECTS OF ANY SERM OR NATURAL ESTROGEN PRESENT...
Please explain. This makes no sense to me (as it would specifically relate to deca), and actually goes against quite a bit of first hand experience with Nolv and Deca. Nolvadex acts as an estrogen in the liver, plain and simple. Estrogenic action in this region of the body stimulates HDL synthesis. How does Deca specifically block out Nolvadex's estrogenic action here? I know androgens play opposing roles, but that is the whole issue. Steroids + Nolvadex to help the prob.. Are you really saying Deca without Nolvadex would be better for HDL than Deca + Nolvadx?
- Bill Llewellyn
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