基于PI3K/Akt信号通路探讨归肾育宫汤对早发性卵巢功能不全大鼠卵巢颗粒细胞自噬的影响  

Effect of Guishen Yugong Decoction on autophagy of ovarian granulosa cells in rat of premature ovarian insufficiency bvia PI3K/AKT signaling pathway

作　　者：卢苑蓉 王强 刘隽阳 许欢[3] 杨鉴冰[4] 符小航[3] LU Yuanrong;WANG Qiang;LIU Junyang;XU Huan;YANG Jianbing;FU Xiaohang(The Second School of Clinical Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China;College of Acupuncture and Massage,Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China;Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China;Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China)

机构地区：[1]陕西中医药大学第二临床医学院,陕西咸阳712046 [2]陕西中医药大学针灸推拿学院,陕西咸阳712046 [3]陕西中医药大学第二附属医院,陕西咸阳712046 [4]陕西中医药大学,陕西咸阳712046

出　　处：《现代中西医结合杂志》2023年第24期3359-3363,3370,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：陕西省自然科学基础研究计划项目(2022JM-491);陕西省中医药管理局科研项目(2021-ZZ-JC016)。

摘　　要：目的基于磷脂酰肌醇3-激酶(PI3K)/丝苏氨酸蛋白激酶(Akt)信号通路探讨归肾育宫汤对早发性卵巢功能不全(POI)大鼠调控卵巢颗粒细胞自噬的机制。方法取清洁级成年健康雌性SD大鼠32只,随机选择8只大鼠作为空白组,其余大鼠给予皮下注射透明带多肽3(pZP3)建立POI模型。将造模成功大鼠随机分为模型组、戊酸雌二醇组、归肾育宫汤组,每组8只。戊酸雌二醇组给予戊酸雌二醇0.09 mg/kg灌胃,归肾育宫汤组给予归肾育宫汤35 g/kg灌胃,模型组和空白组给予等体积生理盐水灌胃,均1次/d。连续灌胃4周后,腹主动脉取血,ELISA法检测血清促卵泡激素(FSH)、雌二醇(E 2)水平;取双侧卵巢称重,计算卵巢指数;取卵巢组织,Western blot法检测卵巢组织中PI3K/Akt信号通路相关蛋白表达情况,免疫组化法观察卵巢颗粒细胞中糖原合酶激酶3(GSK-3)和B淋巴细胞瘤-2(Bcl-2)阳性表达情况。结果与空白组比较,模型组大鼠血清E 2水平、卵巢指数及卵巢组织中Akt、PI3K、3-磷酸肌醇依赖性蛋白激酶1(PDK1)蛋白相对表达量和卵巢颗粒细胞中GSK-3、Bcl-2平均光密度值均明显降低(P均<0.05),血清FSH水平明显升高(P<0.05);与模型组比较,戊酸雌二醇组和归肾育宫汤组大鼠血清E 2水平、卵巢指数及卵巢组织中Akt、PI3K、PDK1蛋白相对表达量和卵巢颗粒细胞中GSK-3、Bcl-2平均光密度值均明显升高(P均<0.05),血清FSH水平均明显降低(P均<0.05),戊酸雌二醇组和归肾育宫汤组各指标比较差异均无统计学意义(P均>0.05)。结论归肾育宫汤可通过调控PI3K/Akt信号通路及其下游相关因子GSK-3、Bcl-2表达,促进卵巢颗粒细胞增殖分化,抑制颗粒细胞过度自噬,调节性激素释放,调控卵泡发育,对POI大鼠卵巢功能起到一定保护作用。Objective It is to explore the mechanism of Guishen Yugong Decoction in regulating autophagy of ovarian granulosa cells in rats with premature ovarian insufficiency(POI)based on the phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(Akt)signaling pathway.Methods Thirty-two clean adult healthy female SD rats were selected,in which 8 ones were randomly selected as the blank group,and the remaining rats were given subcutaneous injection of peptide zona pellucida 3(Zp3)to establish POI models.The successfully modeled rats were randomly divided into model group,estradiol valerate group,Guishen Yugong Decoction group,with 8 rats in each group.The estradiol valerate group was given estradiol valerate 0.09 mg/kg by gavage,the Guishen Yugong Decoction group was given Guishen Yugong Decoction group 35 g/kg by gavage,and the model group and blank group were given an equal volume of normal saline,all once daily.After 4 weeks of continuous gavage,the blood was taken from the rat abdominal aorta,and the serum levels of follicle stimulating hormone(FSH)and estradiol(E 2)were detected by ELISA;both ovaries were weighed to calculate the ovarian index;ovarian tissues were taken,and the expression of proteins related to the PI3K/Akt signaling pathway was detected by Western blot method,and the positive expression of glycogen synthase kinase 3(GSK-3)and B-lymphoblastoma-2(Bcl-2)in ovarian granulosa cells was observed by immunohistochemistry.Results Compared with the blank group,the serum level of E 2,ovarian index and relative expressions of Akt,PI3K,3-phosphatidylinositol-dependent protein kinase 1(PDK1)proteins in ovarian tissues and the average optical density values of GSK-3 and Bcl-2 in ovarian granulosa cells of rats in the model group were all significantly decreased(all P<0.05),and the serum level of FSH was significantly increased(P<0.05);compared with the model group,the serum level of E 2,ovarian index and relative expressions of Akt,PI3K,3-phosphatidylinositol-dependent protein kinase 1(PDK1)proteins in ovarian

关 键 词：早发性卵巢功能不全 卵巢颗粒细胞 磷脂酰肌醇3-激酶 丝苏氨酸蛋白激酶 自噬 

分 类 号：R-332[医药卫生]