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

性别分化和配子发生相关基因生殖功能的分子分析

• 批准号: 14571583
• 负责人: SUEOKA Kou
• 金额: $ 2.24万
• 依托单位: KEIO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571583/
• 关键词: spermatogenesis; doxorubicin; c-kit; F-TRAP; telomerase; TUNEL; green tea extract (GTE); apoptosis; Doxorubicin (DXR); RT-PCR法; mRNA発現; テロメラーゼ活性; c-kit mutant; WBB6F1(wild type)+; +; hetero W^v; W; W^v; Apoptosis

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.

采用RT-PCR和免疫组化方法，在多柔比星（DXR）损伤小鼠睾丸生殖细胞模型中检测c-kit在生殖细胞基因表达和c-kit产生方面的意义。并观察中药TJ41、TJ7和gtc对端粒酶活性和细胞凋亡的促进作用。凋亡细胞用tdt介导的dUDP镍端标记（TUNEL）法进行标记。DXR暴露可使睾丸重量显著降低至55-64%，而gtc与DXR联合处理的参数与对照组相似。在dxr暴露小鼠的睾丸中，对照组睾丸小管中的生殖细胞急剧减少约80%。然而，TJ41和GTCs共同给药可显著减轻生殖细胞损伤。c-Kit及其基因表达在DXR暴露的睾丸中降低，但在促药组中没有。虽然用基于荧光的TRAP方法评估的端粒酶活性在TJ41和gtc的联合使用下与对照组相比有所增加，但在没有这些促进剂的情况下，DXR暴露降低了端粒酶活性。与未使用DXR的促凋亡剂相比，含有DXR的促凋亡剂可显著减少凋亡细胞的数量。这些结果表明，c-kit及其基因表达引起的毒性化学物质暴露后睾丸功能障碍的机制是确定的，TJ41和gtc可能通过抑制睾丸凋亡来促进端粒酶活性，从而对dxr诱导的睾丸毒性具有保护作用。

项目成果

末岡 浩: "遺伝子病の着床前診断(PGD)"産婦人科の世界. 56. 264-270 (2004)

Hiroshi Sueoka：“遗传性疾病的植入前诊断（PGD）”《妇产科世界》56。264-270（2004）。

末岡 浩(分担): "新しい産科学-生殖医療から周産期医療まで"(財)名古屋大学出版会. 283 (2002)

Hiroshi Sueoka（撰稿人）：“新产科 - 从生殖医学到围产期医学”名古屋大学出版社 283（2002）。

末岡 浩: "着床前診断の現状と選択肢"産婦人科の世界. 56(2). 3-10 (2004)

Hiroshi Sueoka：“植入前诊断的现状和选择”《妇产科世界》56(2) (2004)。

末岡 浩(分担): "図説ARTマニュアル"永井書店. 509 (2002)

末冈宏（合伙人）：《插画艺术手册》永井书店 509 (2002)。

Akira Nakabayashi, Kou Sueoka, Noriko Matsuda, Hironori Asada, Reiko Tanigaki, Kenji Satoh, Hiroto Tajima, Tsutomu Ogata, Naoaki Kuji, Yasunori Yoshimura: "Incidental deviation of short and long CAG repeats in the androgen receptor gene for Japanese male

Akira Nakabayashi、Kou Sueoka、Noriko Matsuda、Hironori Asada、Reiko Tanigaki、Kenji Satoh、Hiroto Tajima、Tsutomu Ogata、Naoaki Kuji、Yasunori Yoshimura：“日本男性雄激素受体基因中短和长 CAG 重复的偶然偏差