摘要
目的:基于单磷酸腺苷活化蛋白激酶(AMPK)/沉默调节蛋白1(SIRT1)/沉默调节蛋白3(SIRT3)信号通路研究健脾益肾化浊方治疗多囊卵巢综合征(PCOS)大鼠的机制.方法:8周龄雌性SD大鼠随机分为空白组、模型组、二甲双胍组和健脾益肾化浊方低、中、高剂量组.除空白组外,其余均采用来曲唑灌胃联合高脂饲料构建PCOS模型,造模成功后给与相应药物治疗30 d.HE染色观察动情周期和卵巢组织病理改变;IHC检测大鼠卵巢组织LC3、Beclin-1、AMPK、SIRT1、SIRT3蛋白阳性表达;Western Blot检测大鼠卵巢组织P450、芳香化酶、卵泡抑素、LC3Ⅱ/LC3 Ⅰ、Beclin-1、P62、AMPK、p-AMPK、SIRT1、SIRT3蛋白表达.结果:与模型组比较,健脾益肾化浊方治疗后大鼠动情周期逐渐恢复,卵巢多囊样改变得到改善;卵巢组织P450、LC3Ⅱ/LC3Ⅰ、Beclin-1、AMPK、p-AMPK、SIRT1、SIRT3蛋白表达显著上调(P＜0.05,P＜0.01),芳香化酶、卵泡抑素和P62蛋白表达显著下调(P＜0.05,P＜0.01).结论:健脾益肾化浊方可改善PCOS大鼠卵巢多囊样改变,调节P450、芳香化酶和卵泡抑素表达,促进自噬稳态平衡,其机制可能与AMPK/SIRT1/SIRT3信号通路激活有关.
Abstract
Objective:To explore the mechanism of Jianpi Yishen Huazhuo Formula in the treatment of polycystic ovary syndrome(PCOS)based on adenosine monophosphate activated protein kinase(AMPK)/silencing regulatory protein 1(SIRT1)/silencing regulatory protein 3(SIRT3)signal pathway.Methods:8-week-old female SD rats were randomly divided into blank group,model group,metformin group and low,medium and high dose groups of Jianpi Yishen Huazhuo Formula.In addition to the blank group,the other groups were given letrozole by gavage combined with high-fat diet to build PCOS model,and were given corresponding drugs for 30 days after the model was successfully established.HE staining was used to observe the estrous cycle and pathological changes of ovarian tissue;The positive expression of LC3,Beclin-1,AMPK,SIRT1,SIRT3 protein in the ovarian tissue of rats was detected by IHC;Western Blot was used to detect the expression of P450,Aromatase,Follistain,LC3Ⅱ/LC3 Ⅰ,Beclin-1,P62,AMPK,p-AMPK,SIRT1,SIRT3 proteins in rat ovarian tissue.Results:Compared with the model group,the estrous cycle of rats gradually recovered and the polycystic changes of ovary were improved after the treatment of Jianpi Yishen Huazhuo Formula;The expression of P450,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,AMPK,p-AMPK,SIRT1 and SIRT3 proteins in ovarian tissue were significantly up-regulated(P＜0.05,P＜0.01),and the expression of Aromatase,Follistain and P62 proteins were significantly down-regulated(P＜0.05,P＜0.01).Conclusion:Autophagy homeostasis exists in the ovarian tissue of PCOS rats.Jianpi Yishen Huazhuo Formula can improve the polycystic changes of the ovaries in PCOS rats,regulate the expression of P450,Aromatase and Follistain,and promote the homeostatic balance of autophagy.Its mechanism may be related to the activation of the AMPK/SIRT1/SIRT3 signal pathway.
NETL
NSTL
万方数据