半夏泻心汤加减方治疗多囊卵巢综合征合并胰岛素抵抗的作用及机制

Therapeutic effect and mechanism of Modified Banxia Xiexin Decoction in the treatment of polycystic ovary syndrome complicated with insulin resistance

目的 从磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/葡萄糖转运蛋白4(GLUT4)信号通路探讨半夏泻心汤加减方治疗多囊卵巢综合征(PCOS)合并胰岛素抵抗的作用机制。方法 以来曲唑联合高脂饮食构建PCOS合并胰岛素抵抗大鼠模型。SPF级SD雌性大鼠随机数字表法分为对照组、模型组、二甲双胍组[二甲双胍0.157 5 g/(kg·d)]和半夏泻心高、中、低剂量组[半夏泻心汤加减方26.46、13.23、6.615 g(kg·d)],每组大鼠6只,灌胃相应药液2周。使用血糖仪检测大鼠空腹血糖,HE染色观察卵巢组织病理改变,采用酶联免疫吸附测定大鼠血清性激素、糖脂代谢相关指标,实时荧光定量PCR(RT-PCR)技术、免疫组织化学(IHC)染色法、蛋白质印迹法(Western blotting)分别检测PCOS合并胰岛素抵抗大鼠卵巢组织中PI3K/Akt/GLUT4信号通路相关因子的表达。结果 与对照组相比,模型组大鼠卵泡结构失常,颗粒细胞层数减少,并可见较多囊状扩张卵泡;睾酮(T)、胰岛素抵抗指数(HOMA-IR)、总胆固醇(TC)、甘油三酯(TG)水平升高,差异具有统计学意义(P<0.05);Akt1 mRNA水平和PI3K、p-PI3K、Akt1、p-Akt1、GLUT4蛋白表达水平下降,差异具有统计学意义(P<0.05)。与模型组相比,二甲双胍组、半夏泻心高剂量组、半夏泻心中剂量组、半夏泻心低剂量组可见黄体及不同发育时期的卵泡,且颗粒细胞层数增多、囊状扩张卵泡数目减少甚至消失;二甲双胍组、半夏泻心高剂量组T水平降低,半夏泻心高剂量组TC、TG水平下降,差异具有统计学意义(P<0.05);二甲双胍组、半夏泻心高剂量组、半夏泻心中剂量组、半夏泻心低剂量组PI3K、Akt1 mRNA水平有上升趋势,二甲双胍组、半夏泻心高剂量组、半夏泻心中剂量组GLUT4 mRNA水平有上升趋势,差异无统计学意义(P>0.05);二甲双胍组、半夏泻心高剂量组、半夏泻心中剂量组、半夏泻心低剂量组p-PI3K、GLUT4蛋白表达水平上升,差异具有统计学意义(P<0.05)。结论 半夏泻心汤加减方可改善PCOS合并胰岛素抵抗大鼠胰岛素抵抗、生殖内分泌水平,治疗PCOS,其作用机制与调控PI3K/Akt/GLUT4信号通路相关因子有关。

Objective We aimed to explore the therapeutic effect of Modified Banxia Xiexin Decoction(BXXX)in the treatment of polycystic ovary syndrome(PCOS)complicated with insulin resistance(IR)through regulating the PI3K/Akt/GLUT4 signaling pathway.Methods Letrozole combined with a high-fat diet was used to construct a PCOS and IR rat model.Model rats were divided into 6 groups,namely the control group,the model group,the metformin group[0.1575 g/(kg·d)],and the BXXX high-,medium-,low-dose groups[26.46、13.23、6.615 g(kg·d)],with 6 rats per group,and the corresponding medication was given by gavage for 2 weeks.6 normal rats were taken as the control group.Fasting blood glucose(FBG)levels were measured.Pathological changes in ovarian tissues were observed via HE staining.Serum sex hormone levels,glucose levels,and lipid metabolism indexes were measured by ELISA.The expression levels of PI3K/Akt/GLUT4 signaling pathway-related factors in ovarian tissues of PCOS and IR rats were determined by RT-PCR,IHC staining,and Western blotting.Results Compared with the control group,the structure of the follicles in the model group was abnormal,the number of granular cell layers was reduced,and more cystic dilated follicles were observed.T levels,HOMA-IR levels,and TC and TG levels were increased(P<0.05);and the mRNA levels of Akt1 and the protein expression levels of PI3K,p-PI3K,Akt1,p-Akt1,and GLUT4 were decreased(P<0.05).The corpus luteum and follicles were observed at different developmental stages.Compared with the model group,in the metformin group,BXXX high-dose group,BXXX medium-dose group,and BXXX low-dose group,the number of granulosa cell layers was increased and the number of cystic dilated follicles was decreased or even zero.The T levels in the metformin group and BXXX highdose group and the TC and TG levels in BXXX high-dose group were decreased(P<0.05);and the mRNA levels of PI3K and Akt1 in the metformin group,BXXX high-,medium-,low-dose group were increased(P>0.05).The GLUT4 mRNA level in the metformin group,BXXX high-,medium-dose group was increased(P>0.05).The expression levels of p-PI3K and GLUT4 in the metformin group,BXXX high-,medium-,low-dose group were significantly increased(P<0.05).Conclusion Modified Banxia Xiexin Decoction can improve IR and reproductive endocrine levels in PCOS and IR rats to treat PCOS.Its mechanism is related to the regulation of PI3K/Akt/GLUT4 signaling pathway-related factors.