基于AMPK/SIRT1/SIRT3信号通路探讨健脾益肾化浊方治疗多囊卵巢综合征大鼠的机制  

Discussion on the mechanism of Jianpi Yishen Huazhuo Formula in the treatment of polycystic ovary syndrome rats based on the AMPK/SIRT1/SIRT3 signaling pathway

作　　者：陈苗 马会明[3] 陈冬梅[4] 冯亚宏 张晓静 罗嘉琦 杜小利[1] CHEN Miao;MA Huiming;CHEN Dongmei;FENG Yahong;ZHANG Xiaojing;LUO Jiaqi;DU Xiaoli(The School of Traditional Chinese Medicine of Ningxia Medical University,The Key Laboratory of Integrated Prevention and Treatment of High Incidence in Ningxia Region,Yinchuan 750004,China;Dingbian County Hospital of Traditional Chinese Medicine,Dingbian 718699,China;The School of Basic Medicine of Ningxia Medical University,The Key Laboratory of Fertility Maintenance of the Ministry of Education,Yinchuan 750004,China;The General Hospital of Ningxia Medical University,The Key Laboratory of Stem Cell and Regenerative Medicine,Yinchuan 750004,China;Ningxia Hospital of Traditional Chinese Medicine and Research Institute of Traditional Chinese Medicine,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学中医学院,宁夏区域高发病中西医结合防治研究重点实验室,银川750004 [2]定边县中医院,定边718699 [3]宁夏医科大学基础医学院,生育力保持教育部重点实验室,银川750004 [4]宁夏医科大学总医院,宁夏干细胞与再生医学重点实验室,银川750004 [5]宁夏中医医院暨中医研究院,银川750004

出　　处：《中华中医药杂志》2024年第10期5452-5456,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：宁夏自然科学基金项目(No.2020AAC03430,No.2022AAC03216);国家自然科学基金项目(No.81660813,No.81560812)。

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/LC3Ⅰ、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。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/LC3Ⅰ,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,LC3Ⅱ/LC3Ⅰ,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.

关 键 词：健脾益肾化浊方 多囊卵巢综合征 芳香化酶 卵泡抑素 自噬 AMPK/SIRT1/SIRT3通路 

分 类 号：R285.5[医药卫生—中药学]