新加苁蓉菟丝子汤通过调控AMPK/SIRT1信号通路对卵巢储备功能减退大鼠线粒体功能的影响

Effects of Modified Congrong Tusizi Decoction on mitochondrial function in rats with diminished ovarian reserve via regulating AMPK/SIRT1 signaling pathway

目的研究新加苁蓉菟丝子汤通过调控AMPK/SIRT1信号通路对卵巢储备功能减退(DOR)大鼠线粒体功能的影响。方法大鼠给予雷公藤甲素(500μg/kg)灌胃30 d建立DOR模型,随机分为模型组、调经促孕丸组(1.05 g/kg)、新加苁蓉菟丝子汤组(5.11 g/kg),每组6只,另取6只正常大鼠作为空白组,灌胃给予相应药物。给药2周后,观察大鼠动情周期,HE染色计数卵巢各级卵泡、黄体数,ELISA法检测血清FSH、AMH、INHB水平,透射电镜下观察卵巢组织线粒体结构,流式细胞术检测卵巢组织ROS、MMP水平,试剂盒检测卵巢组织ATP水平,Western blot法检测卵巢组织AMPK、SIRT1、PGC-1α蛋白表达。结果与模型组比较,新加苁蓉菟丝子汤组部分大鼠恢复动情周期,初级、次级卵泡数、黄体数增加(P<0.05,P<0.01),闭锁卵泡数减少(P<0.01);血清FSH水平降低(P<0.05),AMH水平升高(P<0.05);卵巢组织线粒体形态学及功能损伤得到缓解,ROS水平降低(P<0.01),MMP、ATP水平升高(P<0.01);卵巢组织AMPK、SIRT1、PGC-1α蛋白表达升高(P<0.01)。结论新加苁蓉菟丝子汤可有效改善卵巢储备功能减退大鼠线粒体功能障碍,促进卵泡发育,保护卵巢功能,其作用机制可能与激活AMPK/SIRT1信号通路有关。

AIM To investigate the effects of Modified Congrong Tusizi Decoction on mitochondrial function in rats with diminished ovarian reserve(DOR)via AMPK/SIRT1 signaling pathway.METHODS Rats given 30 days triptolide(500μg/kg)by gavage were induced into DOR model and randomly divided into the model group,Tiaojing Cuyun Pills group(1.05 g/kg)and Modified Congrong Tusizi Decoction group(5.11 g/kg),with 6 rats in each group,in contrast to another 6 normal rats in the blank group.After the two weeks corresponding administration,the rats had their estrous cycle observed;their number of follicles and corpus luteum at all levels in ovary counted after HE staining;their levels of serum FSH,AMH and INHB detected by ELISA method;their structure of ovary mitochondria observed under the transmission electron microscope;their levels of ROS and MMP in ovarian tissue detected by flow cytometry;their levels of ATP in ovarian tissue detected by kit;and their protein expressions of AMPK,SIRT1 and PGC-1αin ovarian tissue detected by Western blot.RESULTS Compared with the model group,Modified Congrong Tusizi Decoction group was observed with resumed estrous cycle in some rats;increased number of primary and secondary follicles and corpus luteum(P<0.05,P<0.01);decreased number of atresia follicles(P<0.01);decreased serum FSH level(P<0.05)and increased AMH level(P<0.05);improved morphological and functional damage of mitochondria in ovarian tissue;decreased ROS level(P<0.01);increased MMP and ATP levels(P<0.01);and increased protein expressions of AMPK,SIRT1 and PGC-1αin ovarian tissue(P<0.01).CONCLUSION Modified Congrong Tusizi Decoction can effectively improve mitochondrial dysfunction,promote follicular development and protect ovarian function in DOR rats,and the mechanism may be related to its efficacy in activating AMPK/SIRT1 signaling pathway.