雌马酚对体外培养小鼠睾丸3β-HSD、P450Scc和Vimentin表达的影响

目的探讨睾丸体外培养模型下雌马酚（Equol）暴露对乳鼠睾丸组织结构、睾酮合成相关酶和波形蛋白（Vimentin）的mRNA及蛋白表达影响。方法用浸浴式通气旋转装置体外培养小鼠睾丸组织。实验随机分为DMSO溶剂对照组和4个Equol剂量组（终浓度为0.01、0.10、1.00、10.00 μmol/L），培养72 h。HE染色观察睾丸组织病理形态学变化；实时荧光定量PCR检测睾酮合成相关基因及Vimentin表达水平；免疫组化法检测睾丸组织相关蛋白表达。结果 HE染色结果显示经Equol处理的睾丸组织未观察到明显病理学变化。实时荧光定量PCR结果显示，与DMSO溶剂对照组相比，Equol各剂量组3β羟类固醇脱氢酶（3β-HSD）、细胞色素P450胆固醇侧链裂解酶（P450Scc）、Vimentin mRNA表达差异均无统计学意义(P>0.05)。免疫组化结果显示，与DMSO溶剂对照组相比，3β-HSD、P450Scc、Vimentin的表达在Equol 0.10 μmol/L剂量组上调，10.00 μmol/L剂量组下调。结论 Equol可影响体外培养的小鼠睾丸3β-HSD、P450Scc、Vimentin表达，对睾丸生理功能可能存在潜在的不利影响。

英文摘要:

Objective To investigate the effects of Equol on genes and protein expression of testosterone synthesis related enzymes and Vimentin in testis of perinatal mice in vitro. Methods Testes were isolated and cultured in infiltrating type rotating device for 72 h. The testes were randomly divided into five groups and treated with Equol (DMSO control, 0.01, 0.10, 1.00, 10.00 μmol/L Equol). Morphological changes were observed by HE staining under optical microscope. Expressions of 3β-hydroxysteroid dehydrogenase (3β-HSD),P450 side-chain cleavage enzyme (P450scc), Vimentin were detected by real-time PCR and immunohistochemistry. Results No apparent morphological change in testes was observed compared to control group. The mRNA expression of 3β-HSD, P450Scc, Vimentin show no statistical significance(P>0.05) in all Equol group, while the protein expressions of 3β-HSD, Vimentin, P450scc increased in 0.10 μmol/L and decreased in 10.00 μmol/L Equol group. Conclusion Equol exposure can affect 3β-HSD, P450Scc, Vimentin expression in testes in vitro, means Equol may have potential adverse effects on testosterone production and spermatogenesis.

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