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目的 :探讨黄芩素对小鼠睾丸支持细胞TM4缝隙连接功能的影响及相关机制。方法:MTT法检测黄芩素对TM4细胞的毒性作用;细胞接种荧光示踪法测定TM4细胞间荧光传递功能;Western印迹和细胞免疫荧光法分别检测黄芩素对TM4细胞中Cx43总蛋白及胞膜Cx43蛋白表达的影响。结果:MTT结果显示黄芩素在0~60μmol/L浓度范围内对TM4细胞无毒性作用;细胞接种荧光示踪法结果显示0~20μmol/L的黄芩素可以显著增强TM4细胞荧光传递功能(P<0.01);Western印迹结果表明黄芩素在0~20μmol/L时可以显著增加TM4细胞Cx43蛋白的表达(P<0.01);细胞免疫荧光法表明,0~20μmol/L的黄芩素可以显著增加TM4细胞膜Cx43蛋白表达。结论:在一定浓度范围内(0~20μmol/L),黄芩素能够通过增加小鼠睾丸支持细胞TM4中Cx43蛋白表达而显著增强细胞缝隙连接功能。
Objective: To investigate the effect of baicalein on TM4 gap junctional function in mouse testis support cells and its related mechanism. Methods: Toxicity of baicalein to TM4 cells was detected by MTT assay. Fluorescent tracer method was used to measure the fluorescence transmission of TM4 cells. Western blotting and immunofluorescence were used to detect the expression of Cx43 protein and Cx43 Effect of protein expression. Results: MTT results showed that baicalein had no toxic effect on TM4 cells in the concentration range of 0 ~ 60μmol / L. Fluorescence tracing showed that baicalein at 0 ~ 20μmol / L could significantly enhance the fluorescence transmission of TM4 cells (P < 0.01). The results of Western blotting showed that baicalein could significantly increase the expression of Cx43 protein (P <0.01) at the concentration of 0 ~ 20μmol / L in TM4 cells. Immunofluorescence showed that baicalein at 0 ~ 20μmol / L could significantly increase the expression of TM4 cell membrane Cx43 protein expression. CONCLUSION: Baicalein can significantly enhance the cell gap junctional function by increasing the expression of Cx43 in TM4 cells in a certain concentration range (0 ~ 20μmol / L).