淫羊藿次苷I通过Akt信号通路上调Bmi1促进小鼠精原干细胞增殖

Mechanism of Icariin I promoting proliferation of mouse spermatogenic stem cells

目的:探究淫羊藿次苷I对小鼠精原干细胞增殖的促进作用及相关机制.方法:使用MTT实验、EdU细胞增殖实验检测淫羊藿次苷I对小鼠精原干细胞增殖能力的影响;Western blot法检测淫羊藿次苷I对Bmi1、p-Akt和Akt蛋白的表达情况;RT-PCR实验检测相关基因的表达结果,最后通过抑制剂和siRNA证明p-Akt和Bmi1蛋白之间存在上下游的调控关系.结果:与对照组相比,0.5-1μM淫羊藿次苷I促进小鼠精原干细胞增殖(P<0.05);相比于对照组,0.5-1μM淫羊藿次苷I处理组p-Akt和Bmi1蛋白表达水平上升(P<0.05);Akt抑制剂下调p-Akt和Bmi蛋白水平(P<0.05);siBmi1对Akt mRNA水平无显著影响(P>0.05).结论:淫羊藿次苷I激活Akt信号通路上调Bmi1表达促进小鼠精原干细胞增殖.

Objective The medicine icariin I is applied to mouse spermatogonial stem cells to explore the effect of the medicine on the proliferation function of the cells and the related mechanism for the effect.Methods The changes in the valueadded ability of mouse spermatogenic stem cells in the presence of icariin I were detected by using the MTT and EdU cell proliferation methods.The changes of Bmi1,p-Akt,and Akt proteins in mouse spermatogenic stem cells in the presence of icariin I were detected by Western Blot experiment.The expression of related genes was detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Finally,inhibitors and siRNA were used to explore regulatory relationship between p-Akt and Bmi1 protein.Results Icariin I at the concentration of 0.5-1μM could significantly enhance the proliferation of mouse spermatogenic stem cells as compared with that in the control group(P<0.05).Meanwhile,within the same concentration range,the expression levels of p-Akt and Bmi1 proteins in mouse spermatogenic stem cells were significantly increased under the action of drugs(P<0.05).P-Akt and Bmi1 protein levels were significantly down-regulated in the AKT inhibitor group,and the difference was statistically significant with the control group(P<0.05).There were no significant changes in Akt mRNA levels in the siBmi1 group compared with the control group(P>0.05).Conclusion Icariin I can enhance the proliferation of mouse spermatogonial stem cells,specifically by activating Akt signaling pathway to up-regulate the expression of Bmi1.