Welder A A, Robertson J W, Fugate R D, Melchert R B
University of Oklahoma Health Sciences Center, Department of Toxicology, Colleges of Pharmacy, Oklahoma City 73190, USA.
Toxicol Appl Pharmacol. 1995 Aug;133(2):328-42. doi: 10.1006/taap.1995.1158.
Recent literature reports of myocardial infarction in athletes who self-administer anabolic-androgenic steroid (AAS) and previous animal studies of the effects of AASs on the heart suggest that these drugs may be directly injurious to the myocardium. We have previously demonstrated that 100 microM testosterone cypionate (TC) inhibits all beating activity of primary neonatal rat myocardial cell cultures within 1 hr of exposure and causes significant LDH release by 4 hr of exposure, indicating a direct toxic effect of TC. The purpose of this investigation was to evaluate the effects of commonly abused AASs on primary neonatal rat myocardial cell cultures and to provide insight into early cellular changes that may lead to TC-induced toxicity. Significant LDH release was observed in 5-day-old primary myocardial cell cultures (obtained from 3-to-5-day-old Sprague-Dawley rats) exposed to 100 microM testosterone enanthate (TE), testosterone propionate (TP), and oxymetholone (O) for 4 and 24 hr and in cultures exposed to 100 microM testosterone (T) for 24 hr. Neutral red retention and MTT formazan production were significantly decreased in cell cultures exposed to 100 microM TE, TP, and O after only 4 hr of exposure, indicating a loss of viability and mitochondrial activity. However, there was no effect on viability of cell cultures exposed for 24 hr to 100 microM of a variety of other commonly abused AASs. Phase-contrast microscopy revealed complete disruption of the monolayer in cell cultures treated with 100 microM TE, TP, and O for 4 hr. Treatment of fura-2-loaded myocardial cell cultures with 100 microM TC produced no significant changes in calcium transients or baseline calcium levels for up to 13 min of exposure. These results indicate that O, T, TC, TE, and TP produce a direct toxic effect in heart cell cultures and that early (< 13 min) changes in calcium homeostasis are unlikely to participate in the mechanism of toxicity.
近期有文献报道了自行服用合成代谢雄激素类固醇（AAS）的运动员发生心肌梗死的情况，并且先前有关AAS对心脏影响的动物研究表明，这些药物可能会直接损害心肌。我们之前已经证明，100微摩尔环丙孕酮睾酮（TC）在暴露1小时内可抑制原代新生大鼠心肌细胞培养物的所有跳动活动，并在暴露4小时后导致显著的乳酸脱氢酶（LDH）释放，这表明TC具有直接毒性作用。本研究的目的是评估常见滥用AAS对原代新生大鼠心肌细胞培养物的影响，并深入了解可能导致TC诱导毒性的早期细胞变化。在5日龄原代心肌细胞培养物（从3至5日龄的Sprague-Dawley大鼠获取）中，暴露于100微摩尔庚酸睾酮（TE）、丙酸睾酮（TP）和羟甲烯龙（O）4小时和24小时后，以及暴露于100微摩尔睾酮（T）24小时后，观察到显著的LDH释放。仅在暴露4小时后，暴露于100微摩尔TE、TP和O的细胞培养物中中性红保留率和MTT甲臜生成显著降低，表明细胞活力和线粒体活性丧失。然而，暴露于100微摩尔的多种其他常见滥用AAS 24小时对细胞培养物的活力没有影响。相差显微镜显示，用100微摩尔TE、TP和O处理4小时的细胞培养物中单层细胞完全破坏。用100微摩尔TC处理负载fura-2的心肌细胞培养物，在长达13分钟的暴露时间内，钙瞬变或基线钙水平没有显著变化。这些结果表明，O、T、TC、TE和TP在心脏细胞培养物中产生直接毒性作用，并且钙稳态的早期（<13分钟）变化不太可能参与毒性机制。
