罗格列酮对大鼠睾丸间质细胞类固醇生成的慢性作用：体内和体外研究。

Effect of chronic treatment with Rosiglitazone on Leydig cell steroidogenesis in rats: in vivo and ex vivo studies.

机构信息

Department of Morphology and Physiology, Universidade Federal Rural de Pernambuco, Recife, 52.171-900, Brazil.

出版信息

Reprod Biol Endocrinol. 2010 Feb 9;8:13. doi: 10.1186/1477-7827-8-13.

DOI:10.1186/1477-7827-8-13

PMID:20144211

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2829566/

Abstract

BACKGROUND

The present study was designed to examine the effect of chronic treatment with rosiglitazone - thiazolidinedione used in the treatment of type 2 diabetes mellitus for its insulin sensitizing effects - on the Leydig cell steroidogenic capacity and expression of the steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage enzyme (P450scc) in normal adult rats.

METHODS

Twelve adult male Wistar rats were treated with rosiglitazone (5 mg/kg) administered by gavage for 15 days. Twelve control animals were treated with the vehicle. The ability of rosiglitazone to directly affect the production of testosterone by Leydig cells ex vivo was evaluated using isolated Leydig cells from rosiglitazone-treated rats. Testosterone production was induced either by activators of the cAMP/PKA pathway (hCG and dbcAMP) or substrates of steroidogenesis [22(R)-hydroxy-cholesterol (22(R)-OH-C), which is a substrate for the P450scc enzyme, and pregnenolone, which is the product of the P450scc-catalyzed step]. Testosterone in plasma and in incubation medium was measured by radioimmunoassay. The StAR and P450scc expression was detected by immunocytochemistry.

RESULTS

The levels of total circulating testosterone were not altered by rosiglitazone treatment. A decrease in basal or induced testosterone production occurred in the Leydig cells of rosiglitazone-treated rats. The ultrastructural and immunocytochemical analysis of Leydig cells from rosiglitazone-treated rats revealed cells with characteristics of increased activity as well as increased StAR and P450scc expression, which are key proteins in androgen biosynthesis. However, a number of rosiglitazone-treated cells exhibited significant mitochondrial damage.

CONCLUSION

The results revealed that the Leydig cells from rosiglitazone-treated rats showed significant reduction in testosterone production under basal, hCG/dbcAMP- or 22 (R)-OH-C/pregnenolone-induced conditions, although increased labeling of StAR and P450scc was detected in these cells by immunocytochemistry. The ultrastructural study suggested that the lower levels of testosterone produced by these cells could be due to mitochondrial damage induced by rosiglitazone.

摘要

背景

本研究旨在探讨用于治疗 2 型糖尿病的罗格列酮 - 噻唑烷二酮（一种胰岛素增敏剂）的慢性治疗对正常成年大鼠睾丸间质细胞甾体生成能力和甾体急性调节蛋白（StAR）和胆固醇侧链裂解酶（P450scc）表达的影响。

方法

将 12 只成年雄性 Wistar 大鼠用罗格列酮（5mg/kg）经口灌胃 15 天进行治疗。另外 12 只对照动物用载体进行治疗。使用来自罗格列酮处理大鼠的分离的睾丸间质细胞，评估罗格列酮直接影响睾丸间质细胞产生睾酮的能力。通过激活 cAMP/PKA 途径的激动剂（hCG 和 dbcAMP）或甾体生成的底物[22(R)-羟基胆固醇（22(R)-OH-C），这是 P450scc 酶的底物，和 pregnenolone，这是 P450scc 催化步骤的产物]诱导睾酮产生。用放射免疫法测量血浆和孵育培养基中的睾酮。用免疫细胞化学检测 StAR 和 P450scc 的表达。

结果

罗格列酮处理并未改变总循环睾酮水平。基础或诱导的睾酮产生在罗格列酮处理大鼠的睾丸间质细胞中减少。罗格列酮处理大鼠的睾丸间质细胞的超微结构和免疫细胞化学分析显示，细胞具有增加的活性以及增加的 StAR 和 P450scc 表达的特征，这是雄激素生物合成的关键蛋白。然而，许多罗格列酮处理的细胞表现出明显的线粒体损伤。

结论

结果表明，罗格列酮处理大鼠的睾丸间质细胞在基础、hCG/dbcAMP 或 22(R)-OH-C/pregnenolone 诱导条件下显示出显著减少的睾酮产生，尽管这些细胞中的 StAR 和 P450scc 的免疫细胞化学标记增加。超微结构研究表明，这些细胞产生的较低水平的睾酮可能是由于罗格列酮诱导的线粒体损伤。