桃核承气汤对多囊卵巢综合征大鼠卵巢组织PI3K/AKT/mTOR信号通路的影响

Effects of Taohe Chengqi Decoction on PI3K/AKT/mTOR signaling pathway in ovary tissue of rats with polycystic ovary syndrome

目的探究桃核承气汤对多囊卵巢综合征(PCOS)大鼠卵巢组织磷脂酰肌醇3-激酶(PI3K)/丝氨酸/苏氨酸蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响。方法将96只雌性SD大鼠随机分为对照组,PCOS组,二甲双胍组,桃核承气汤低、中、高剂量组,每组16只;除对照组外,其余各组大鼠皮下注射脱氢表雄酮构建PCOS模型。造模成功后二甲双胍组大鼠给予0.1 g/kg二甲双胍灌胃,桃核承气汤低、中、高剂量组分别给予1.89、3.78、7.56 g/kg桃核承气汤灌胃,对照组和PCOS组灌胃等量生理盐水,1次/d,共28 d。血糖仪检测空腹血糖(FPG)水平,酶联免疫吸附试验检测血清卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E_(2))、睾酮(T)、空腹胰岛素(FINS)水平,计算胰岛素抵抗指数(HOMA-IR);苏木素伊红染色观察大鼠卵巢组织病理学情况;荧光定量PCR和蛋白免疫印迹实验分别检测大鼠卵巢组织PI3K/AKT/mTOR通路相关分子mRNA和蛋白表达。结果与对照组相比,PCOS组大鼠卵泡体积增大,含卵泡液,且细胞间隙增大、黄体减少、泡膜细胞层增生,血清FSH、E_(2),卵巢组织中PI3K/AKT/mTOR通路mRNA和蛋白表达降低,LH、T、FPG、FINS及HOMA-IR水平升高(P<0.05);与PCOS组相比,二甲双胍组及桃核承气汤低、中、高剂量组大鼠卵泡体积减小、黄体数目增加、泡膜细胞层增生现象减轻,血清FSH、E_(2),卵巢组织中PI3K/AKT/mTOR通路mRNA和蛋白表达升高,LH、T、FPG、FINS及HOMA-IR水平降低(P<0.05);桃核承气汤低、中、高剂量组上述指标变化呈剂量依赖性,低、中剂量组与二甲双胍组差异有统计学意义,高剂量组与二甲双胍组差异无统计学意义。结论桃核承气汤可促进PCOS模型大鼠性激素恢复正常水平,抑制胰岛素抵抗,这可能与其促进PI3K/AKT/mTOR信号通路激活有关。

Objective To explore the effect of Taohe Chengqi Decoction on phosphatidylinositol 3-kinase(PI3K)/serine/threonine protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway in ovarian tissue of a rat model of polycystic ovary syndrome(PCOS).Methods Ninety-six female SD rats were randomly divided into the control group,the PCOS group,the metformin group,the Taohe Chengqi Decoction low,medium and high dose groups,with 16 rats in each group.Except for the control group,the PCOS model was constructed by subcutaneous injection of dehydroepiandrosterone in the other groups.After successful model building,the metformin group was given 0.1 g/kg metformin by gavage,and the Taohe Chengqi Decoction low,medium and high dose groups were given 1.89 g/kg,3.78 g/kg and 7.56 g/kg Taohe Chengqi Decoction by gavage,respectively.The control group and the PCOS group were given the same amount of normal saline by gavage,1 time/d for 28 days.The fasting blood glucose(FPG)level was measured by blood glucose meter.Enzyme linked immunosorbent assay was used to detect serum levels of follicle stimulating hormone(FSH),luteinizing hormone(LH),estradiol(E_(2)),testosterone(T)and fasting insulin(FINS).Insulin resistance index(HOMA-IR)was calculated.Hematoxylin eosin staining was used to observe the histopathology of rat ovary.The mRNA and protein expression of PI3K/AKT/mTOR pathway in rat ovarian tissue were detected by fluorescence quantitative PCR and Western blot assay.Results Compared with the control group,the volume of follicles increased,contained follicular fluid,and the cell space increased,corpus luteum decreased,and the cell layer of theca proliferated,serum FSH,E_(2),mRNA and protein expression of PI3K/AKT/mTOR pathway in ovarian tissue were decreased,and levels of LH,T,FPG,FINS and HOMA-IR were increased in the PCOS group(P<0.05).In the metformin group and the Taohe Chengqi Decoction low-dose,medium-dose and high-dose groups,the volume of follicles decreased,the number of corpus luteum increased,and the proliferation of alveolar cell layer decreased.Serum levels of FSH,E_(2),mRNA and protein expression of PI3K/AKT/mTOR pathway in ovarian tissue were increased,and levels of LH,T,FPG,FINS and HOMA-IR were decreased(P<0.05).Changes of above indexes were dose-dependent in the low,medium and high dose groups of Taohe Chengqi Decoction.There were significant differences in changes of above indexes between the Taohe Chengqi Decoction low,medium-dose groups and the metformin group.There was no significant difference between the high-dose of Taohe Chengqi Decoction group and the metformin group.Conclusion Taohe Chengqi Decoction can restore normal levels of sex hormones and inhibit insulin resistance in PCOS model rats,which may be related to its promotion of PI3K/AKT/mTOR signal pathway activation.