Testosterone and CholesterolOf all the side effects associated with high-dose anabolic steroid use, the most immediately dangerous is the way they do anabolic steroids raise cholesterol the cardiovascular system. Although a direct raisee relationship has never been established between steroid use and cardiovascular anabloic, many cases published in the medical literature point an accusatory finger at injudicious steroid use as being the how to take liquid tren cause of death from CVD. Taking steroids can lead to an enlarged heart that goes beyond that induced by exercise alone. That can set you up for future heart failure. The immediate cause of most heart attacks is an internal clot that obstructs do anabolic steroids raise cholesterol coronary artery. Steroids come into play because they promote internal clotting.
Cholesterol is dangerously high
There is a comprehensive body of evidence documenting that anabolic-androgenic steroids are associated with deleterious alterations of lipid profile. In this study we investigated whether a single dose of testosterone enanthate affects the cholesterol biosynthesis and the expression of HMGCR.
The total cholesterol levels prior to and two days after testosterone administration were analysed. This is the first time a perturbation in the lipoprotein profile is observed after only a single dose of testosterone.
Here we provide a molecular explanation how anabolic androgenic steroids may impact on the cholesterol homeostasis, i.
Increasing knowledge and understanding of AAS induced side-effects is important in order to find measures for treatment and care of these abusers. Anabolic androgenic steroids AAS including testosterone, other endogenous androgenic hormones and synthetic substances structurally related to these compounds are the most frequently detected doping agents in the society and sports.
The abuse of these agents for cosmetic purposes among non-competitive recreational body-builders and non-athletes is a considerable health concern. There is a comprehensive body of evidence documenting that AAS induce various deleterious alterations of the lipoprotein profile.
The most prominent changes include elevations of low density lipoprotein LDL and decreases of high density lipoprotein HDL [ 4 - 7 ]. The long-term consequences of these alterations are still unknown but it is possible that the perturbation of the lipid profile may be associated with an increase in risk of coronary artery disease.
Competitive inhibitors of the HMGCR by statins lead to induction of the expression of LDL receptors in the liver, which in turn increases the catabolism of plasma LDL and lowers the concentration of cholesterol in plasma. It is conceived that statins have a preventive effect on cardiovascular disease to a great extent by these mechanisms in several populations [ 8 ].
In this study we investigated whether a single dose of testosterone enanthate affects the cholesterol profile and the expression of HMGCR in healthy volunteers. The lipoprotein profile was analysed prior to, and two and fifteen days after administration of mg testosterone enanthate. Study subjects included 39 healthy volunteers originating from the study population described in detail elsewhere [ 9 ]. All participants were males at age years, and gave informed consent consistent with the approval of the Ethics Review Board.
Blood and serum was collected prior to day 0 , 2 and 15 days after testosterone administration. Averse drug reactions ADRs were monitored from the time of screening until day 15 after administration of testosterone. Frozen whole blood samples were available from 24 subjects of the 39 individuals included in the study. The blood samples were mixed 1: A standard curve with increasing concentration of liver microsomes was included on each gel.
Prior to testosterone treatment the HepG2 cells were split and plated in 6-well plates and pre-incubated for 3 days. The non-treated controls were incubated with vehicle only. The experiments were performed in four independent experiments. Total RNA extraction was performed using 0.
Each reaction was performed in triplicates and no-template controls were included in each experiment. The gene expression was quantified as the yield of the target gene relative to that of Beta-actin gene.
Total testosterone levels in serum prior to and after testosterone administration had been measured by GCMS in a previous study [ 12 ]. Statistical analyses were performed using GraphPad Prism software 4. Total cholesterol level increased on day two mean 4. On day 15 the total cholesterol level was back to baseline mean 4. The change in total cholesterol levels on day 0 prior to testosterone administration , 2 and 15 days after testosterone administration.
The increase in cholesterol level prompted us to investigate the expression of the main enzyme involved in the cholesterol synthesis. The protein expression of HMGCR, the rate limiting enzyme in the cholesterol synthesis, was investigated by Western blotting in whole blood from the subjects prior to, and two days after testosterone administration. HMGCR protein levels in vivo. The individual expression values on day 2 were presented in relation to the level on day 0 which was set to one. HepG2 cells were exposed to testosterone enanthate 1 uM for hours.
After 48 hours the mRNA levels were back to basal level. Even though accumulating evidence indicates that testosterone may have adverse effect on the lipoprotein profile and cardiovascular health, this is, to the best of our knowledge, the first time an increase in total cholesterol level has been observed after only one single dose of testosterone.
Numerous reports on the effects of AAS on lipoproteins in humans have been published in the last 25 years. To a large extent the results of these studies present consistent results, i. However, for the effect on total cholesterol levels the results are conflicting. Some studies have found that repeated supra-physiologic doses of AAS is associated with an increase in total cholesterol levels [ 15 , 16 ], whereas others have failed to find such an association [ 5 , 6 ].
The reason for the discrepancy observed in the effect on total cholesterol after AAS administration may be the different study designs used, sampling time, type of AAS used, administration route etc. However, most importantly, these studies all concerned subjects with multiple doses or chronic use of AAS. Here we show that circulatory concentration of total testosterone was significantly associated with the increase in cholesterol levels on day 2. No toxic effect on liver function, as assessed by ALAT and ASAT, was found indicating that the increase in total cholesterol observed is directly associated with the administration of testosterone and not an artificial effect of the injection, i.
The synthesis of cholesterol is dependent on the activity of HMGCR, and we therefore investigated if testosterone could affect the expression of this enzyme. Here we show for the first time that a supra-physiological dose of testosterone induces the expression of HMGCR in vivo in healthy volunteers. This concentration is in the range of the levels achieved after administration of testosterone to the volunteers [ 12 ], and yielded a marked induction of the transcription of the HMGCR gene.
Further support for this testosterone effect is provided from experiments in castrated mice and male Sprague-Dawley rats in which 14 days administration of AAS testosterone and nandrolone led to an up-regulation of meibomian and adrenal gene expression of HMGCR, respectively [ 18 , 19 ]. The mechanisms for the androgen induced up-regulation of HMGCR transcription as well as the physiological consequences have not been investigated and needs to be further elucidated.
High cholesterol levels are known to exert a negative feedback on the cholesterol synthesis on a transcriptional level [ 20 , 21 ]. This may explain the time dependent response observed in our HepG2 experiments, i. Other molecular mechanisms behind the unfavourable effects of AAS on the lipoprotein profile have not been well investigated.
Here we provide an additional molecular explanation how AAS may impact on the cholesterol homeostasis, i. Further studies may provide yet other explanations behind the lipid profile perturbation observed as a result of AAS abuse. AAS are the most commonly used doping agents in athletes http: However, the abuse of AAS is not limited to competitive sports, but has spread to sport amateurs and non-athletes and is considered a serious concern in the society [ 2 , 22 ].
It is therefore urgent to study and establish the awareness of the adverse effects caused by AAS. Here we provide scientific evidence that a supra-physiological dose of testosterone may have adverse effects on the cholesterol metabolism. Our setting is, however, very different from the situation for the illicit users that typically take AAS in repeated courses known as "cycles", each lasting several weeks to several months.
One might suspect that the HMGCR expression will rapidly increase after each dose, keeping the HMGCR protein expression and cholesterol metabolism elevated continuously during the "cycles". Some limitations of our study need to be addressed.
Using whole blood as surrogate model for HMGCR expression may not reflect the expression profile in the liver. Nevertheless, blood is a natural surrogate organ in the absence of availability of liver biopsies. SREBP-2 are highly abundant in the blood cells further supporting that whole blood may be used for expression analysis.
Another limitation is that we only have the cholesterol profile in our study participants. The inclusion of cholesterol activity biomarkers such as lathosterol would further support that the administration of supra-physiological doses of testosterone disturb the cholesterol metabolism in vivo. Unfortunately our study design did not permit further analysis of additional biomarkers.
In conclusion, we have shown that already one single dose of testosterone enanthate increases the serum total cholesterol level. This immediate response to AAS is a cause for concern and signals that the lipid metabolic perturbation is a rapid response and warrants close follow-up of the cardiovascular risk factors that may appear later in life in abusers. There are many cases described in the literature on such adverse events in heavy abusers of AAS.
Our findings demonstrate that such effects may occur also in subjects with moderate, intermittent, or temporary abuse of AAS. Therefore, given our findings, public efforts should be centered on primary prevention. LE and AR planned, designed and wrote the study protocol for the in vivo study. CS planned and performed the cell culture analysis. LE and NG interpreted the data. NG performed the statistical analysis and managed the sample collection. LE wrote the first draft of the manuscript. All authors contributed to and have approved the final manuscript.
The technical assistance of Birgitta Ask is gratefully acknowledged. National Center for Biotechnology Information , U. Subst Abuse Treat Prev Policy. Published online Mar Received Nov 7; Accepted Mar This article has been cited by other articles in PMC. Conclusion Here we provide a molecular explanation how anabolic androgenic steroids may impact on the cholesterol homeostasis, i.
Background Anabolic androgenic steroids AAS including testosterone, other endogenous androgenic hormones and synthetic substances structurally related to these compounds are the most frequently detected doping agents in the society and sports.
Methods Subjects and design Study subjects included 39 healthy volunteers originating from the study population described in detail elsewhere [ 9 ]. Data analysis Statistical analyses were performed using GraphPad Prism software 4. Results Cholesterol levels Total cholesterol level increased on day two mean 4.
Open in a separate window. HMGCR expression in whole blood The increase in cholesterol level prompted us to investigate the expression of the main enzyme involved in the cholesterol synthesis.
Discussion Even though accumulating evidence indicates that testosterone may have adverse effect on the lipoprotein profile and cardiovascular health, this is, to the best of our knowledge, the first time an increase in total cholesterol level has been observed after only one single dose of testosterone. Conclusions In conclusion, we have shown that already one single dose of testosterone enanthate increases the serum total cholesterol level. Competing interests The authors declare that they have no competing interests.
Authors' contributions LE and AR planned, designed and wrote the study protocol for the in vivo study.