小檗碱对多囊卵巢综合征大鼠卵巢局部胰岛素抵抗的调控作用  

Regulatory Effect of Berberine on Local IR in Ovary of PCOS Rats Based on AMPK/mTOR Signaling Pathway

作　　者：张多加[1] 张明明[2] 王婷婷[2] 张春兰[1] 郭滢[1] 吴效科[1] 李慕白[1] ZHANG Duojia;ZHANG Mingming;WANG Tingting;ZHANG Chunlan;GUO Ying;WU Xiaoke;LI Mubai(Department of Obstetrics and Gynecology,First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院妇一科,哈尔滨150040 [2]黑龙江中医药大学,哈尔滨150040

出　　处：《世界中医药》2022年第17期2429-2436,共8页World Chinese Medicine

基　　金：黑龙江省自然科学基金联合引导项目(LH2019H116)——基于AMPK-TSC-mTOR通路研究黄连素对PCOS卵巢胰岛素抵抗及DNA甲基化的调控。

摘　　要：目的:研究小檗碱对多囊卵巢综合征(PCOS)大鼠卵巢局部AMP活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)靶点的调控作用,从而改善卵巢局部胰岛素抵抗的作用机制。方法:选取60只清洁型雌性Sprague-Dawley(SD)大鼠,随机分为空白组、模型组、小檗碱组、二甲双胍组,每组15只。除空白组外,给予生物合成人胰岛素注射液(NS)联合注射用人绒毛膜促性腺激素(HCG)建立PCOS大鼠模型。小檗碱组、二甲双胍组分别给予小檗碱、二甲双胍灌胃。干预结束后,检测各组大鼠血清性激素及糖脂代谢水平;观察卵巢形态学;蛋白质印迹法(Western Blotting)检测卵巢组织磷酸化AMP活化蛋白激酶苏氨酸-172α(α-pT172-AMPK)、磷酸化哺乳动物雷帕霉素靶蛋白丝氨酸-2481(pSer2481-mTOR)、磷酸化蛋白激酶B丝氨酸-473(pSer473-AKT)蛋白表达水平;实时荧光聚合酶链式反应法(PCR)检测卵巢组织中葡萄糖转运蛋白4(GLUT4)信使RNA表达水平。结果:与空白组比较,模型组大鼠血清性激素中总睾酮(T)、黄体生成素(LH)、LH/促卵泡素(FSH)、17α-羟孕酮(17α-OHP)和雄烯二酮(ASD)水平明显升高,性激素结合球蛋白(SHBG)水平明显下降;空腹胰岛素(FINS)和稳态模型胰岛素抵抗指数(HOMA-IR)水平均明显升高;卵巢形态学观察显示,囊性扩张卵泡多见,各发育期卵泡减少,卵泡颗粒细胞层数减少;卵巢组织中α-pT172-AMPK和pSer473-AKT水平明显降低,pSer2481-mTOR蛋白表达水平明显升高,GLUT4 mRNA表达水平明显降低。与模型组比较,小檗碱组大鼠血清生殖代谢指标有明显改善;卵巢形态学恢复;卵巢组织中α-pT172-AMPK和pSer473-AKT水平明显升高,pSer2481-mTOR蛋白表达水平明显降低,GLUT4 mRNA表达水平明显升高。结论:小檗碱能够改善PCOS大鼠生殖内分泌紊乱,调节胰岛素抵抗,恢复卵巢组织中卵泡发育,其机制与小檗碱作用于AMPK/mTOR靶点,调控卵巢局部胰岛素信号通路转导障Objective:To investigate the regulatory effect of berberine on local AMP-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR) targets of ovaries in rats with polycystic ovary syndrome(PCOS) and thereby to explore the mechanism of berberine in improving local insulin resistance(IR) of ovary.Methods:A total of 60 female Sprague-Dawley(SD) rats were randomized into blank group, model group, berberine group, and metformin group, with 15 rats in each group.The Biosynthetic Human Insulin Injection(NS) and human chorionic gonadotropin(HCG) were used to induce PCOS in rats except for the blank group.Berberine group and metformin group were given(ig) berberine and metformin, separately.After the intervention, serum sex hormone levels and glucose and lipid metabolism were detected in each group, and the morphological changes of ovary were observed.The protein expression of α-pT172-AMPK,pSer2481-mTOR,and pSer473-AKT in ovary was detected by Western Blotting, and the mRNA expression of glucose transporter 4(GLUT4) in ovary by real-time fluorescent quantitative PCR.Results:1) Compared with the blank group, the model group showed significantly high levels of the hormones serum total testosterone(T),luteinizing hormone(LH),LH/follicle-stimulating hormone(FSH),17α-hydroxyprogesterone(17α-OHP),and androstenedione(ASD),significantly low level of sex hormone binding globulin(SHBG),and significantly high levels of fasting insulin(FINS) and homeostasis model assessment of insulin resistance(HOMA-IR).The ovary showed many cystic follicles and reduction in the number of follicles and granulosa cell layers of follicles at each development stage.Moreover, significant decrease in the levels of α-pT172-AMPK and pSer473-AKT and the mRNA expression of GLUT4 and significant increase in the protein level of pSer2481-mTOR in the ovary were observed in the model group as compared with those in the blank group.2) Compared with the model group, berberine group demonstrated significant improvement in serum reproductive metabolism

关 键 词：多囊卵巢综合征 胰岛素抵抗 小檗碱 AMP活化蛋白激酶 哺乳动物雷帕霉素靶蛋白 

分 类 号：R285.5[医药卫生—中药学]