Molecular analysis of reproductive fanction on the genes implicated to sex differentiation and gametogenesis

以阿霉素（DXR）损伤小鼠睾丸作为生殖细胞毒性模型，采用RT-PCR和免疫组化方法，从基因表达和c-kit产生方面探讨c-kit在睾丸生殖细胞中的意义。同时观察中药（TJ 41、TJ 7）和GTC对端粒酶活性和细胞凋亡的促进作用。通过TdT介导的dUDP缺口末端标记（TUNEL）测定来标记凋亡细胞。DXR暴露使睾丸重量显著降低至55-64%，而GTC与DXR的组合处理导致与对照相似的参数。在DXR暴露小鼠的睾丸中，通过组织学分析，对照组睾丸小管中的生殖细胞急剧减少约80%。然而，生殖细胞损伤显着减弱TJ 41和GTC共同管理。DXR组睾丸c-Kit及其基因表达降低，促性腺激素组无明显变化。与对照组相比，TJ 41和GTC联合应用后，TRAP法检测的端粒酶活性明显升高，而DXR法检测的端粒酶活性则明显降低。与没有这些试剂的DXR暴露相比，具有DXR的促进剂显示出凋亡细胞数量的显著减少。这些结果表明，睾丸功能障碍暴露后的有毒化学品是确定的机制引起的c-kit及其基因表达和TJ 41和GTC可能有重大的证据，对DXR诱导的睾丸毒性，通过促进端粒酶活性抑制睾丸细胞凋亡的保护作用。

The implication of c-kit was examined on the aspect of gene expression and c-kit production at testicular germ cell by RT-PCR and immunohistochemistry in the mice testes damaged by doxorubicin (DXR) as a germ cell toxic model. And also the promoting effect of herbal medicine (TJ41, TJ7) and GTCs on telomerase activity and apoptosis were investigated. Apoptotic cells were labelled by the TdT-mediated dUDP nick-end labelling (TUNEL) assay. The weight of testes was significantly decreased to 55-64% by DXR exposure, while the combined treatment of GTCs with DXR resulted in parameters similar to the control. In the testes of DXR-exposed mice, germ cells in the testicular tubules of control were drastically reduced by about 80% by histological analysis. However, germ cell damage was significantly attenuated by TJ41 and GTCs coadministration. c-Kit and its gene expression were reduced in the DXR exposured testis, but not in the group with promoting agents. Although the telomerase activity evaluated by fluorescence-based TRAP assay was increased with coadministration of TJ41 and GTCs compared with control, that was decreased by DXR expoosure without these promoting agents. The promoting agents with DXR show a marked reduction in the number of apoptotic cells compared with DXR exposure without these agents. These results suggest that testicular dysfunction following exposure of toxic chemicals are determining at the mechanisms provoked by c-kit and its gene expression and TJ41 and GTCs may have eventful evidence of a protective effect against DXR-induced testicular toxicity via promoting telomerase activity by inhibition of testicular apoptosis.