东南大学附属中大医院
※基金项目 国家自然科学基金（No.81603623；No.81874472）
目的 研究养精胶囊促进小鼠精原干细胞（SSCs）增殖的分子机制。方法 将不同浓度的养精胶囊提取物加入SSCs中培养48 h，CCK-8检测细胞的增殖活性，流式细胞仪检测细胞周期，荧光素酶报告基因检测Cyclin D1启动子的活性，qRT-PCR以及免疫荧光检测Cyclin D1的表达。之后进行阻断实验，在加入养精胶囊前预先加入siRNA-Cyclin D1，同样的方法检测细胞的增殖活性、细胞周期、Cyclin D1启动子的活性以及Cyclin D1的表达情况。结果 低、中、高浓度的养精胶囊可以促进SSCs的增殖，提高S期细胞的比例，增强Cyclin D1启动子的活性，促进Cyclin D1的表达。阻断Cyclin D1后，SSCs的增殖活性降低，S期细胞比例减少，Cyclin D1启动子的活性降低，Cyclin D1的表达减少。结论 养精胶囊通过增强Cyclin D1启动子的活性提高Cyclin D1的转录和翻译水平，进而促进SSCs增殖。
Objective: To study the molecular mechanism of Yangjing Capsule (YC) promoting the proliferation of mouse spermatogonia stem cells (SSCs). Methods: YC extract of different concentrations were added to SSCs for 48 h, and the proliferation activity of cells was detected by CCK-8, the cell cycle was detected by flow cytometry, the activity of Cyclin D1 promoter was detected by luciferase reporter assay, and the expression of Cyclin D1 was detected by qRT-PCR and immunofluorescence. Then the blocking experiments were carried out, siRNA-Cyclin D1 was added before the treatment of YC, and the proliferation activity, the cell cycle of cells, as well as the activity of Cyclin D1 promoter and the expression of Cyclin D1 were detected by the same methods. Results: The low, medium and high dose of YC could promote the proliferation of SSCs, increase the proportion of S-stage cells, enhance Cyclin D1 promoter activity, and promote the expression of Cyclin D1. Blocking experiments showed that pre-treatment of siRNA-Cyclin D1 could reduce the proliferation activity of SSCs, reduce the proportion of S-phase cells, decrease Cyclin D1 promoter activity, and inhibit the expression of Cyclin D1. Conclusion: YC extract promotes proliferation of mouse spermatogonia stem cells through regulating Cyclin D1 promoter and improving the transcription and translation of Cyclin D1.
蔡滨,金保方,孙大林,邓伟民. Cyclin D1启动子在养精胶囊促进小鼠精原干细胞增殖中的作用※[J].,2020,19(6):70-74
复制
