当归多糖对多囊卵巢综合征大鼠氧化应激状态改善的机制

Mechanism of Angelica sinensis polysaccharide on the improvement of oxidative stress in polycystic ovary syndrome rats

目的探讨当归多糖(Angelica sinensis polysaccharide,ASP)对多囊卵巢综合征大鼠氧化应激状态的改善作用及其可能的作用机制。方法选取48只雌性大鼠,用来曲唑灌胃法建立多囊卵巢综合征(polycystic ovary syndrome,PCOS)模型,将造模成功的大鼠随机分为模型组、ASP低剂量组、ASP低高剂量组和西药组,各12只。另设对照组(12只)。ASP低剂量组、ASP高剂量组灌胃给药50、200 mg·kg^(-1)ASP(用生理盐水配制),西药组灌胃给药200 mg·kg^(-1)二甲双胍溶液,对照组及模型组给予等量生理盐水,连续给药28 d。酶联免疫吸附试验(enzyme linked immunosorbent assy,ELISA)测定各组大鼠血清中雌二醇(estradiol,E 2)、睾酮(testosterone,T)、促卵泡生成素(follicle stimulating hormone,FSH)、促黄体生成素(luteinizing hormone,LH)和丙二醛(malondialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和超氧化物岐化酶(superoxide dismutase,SOD)水平;测定卵巢质量和卵巢体积并分析卵巢形态变化;HE染色观察各组大鼠卵巢组织病理变化;实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)和蛋白质印迹法(Western blotting)分别检测卵巢组织中核因子E2相关因子2(nuclearfactor erythroidderived 2-like 2,Nrf2)和血红素加氧酶1(heme oxygenase-1,HO-1)mRNA和蛋白的表达情况。结果与模型组比较,3个给药组大鼠血清E 2、FSH、GSH-Px、SOD、Nrf2、HO-1 mRNA和蛋白的表达水平升高,T、LH、MDA、卵巢质量和卵巢体积减小(P<0.05)。与ASP低剂量组比较,ASP高剂量组和西药组各项指标改变更明显(P<0.05)。结论ASP能调整PCOS大鼠血清氧化因子水平,减轻氧化应激反应,改善卵巢囊样扩张,可能是通过激活Nrf2/HO-1信号通路发挥作用的。

Objective To investigate the effect of Angelica sinensis polysaccharide(ASP)on oxidative stress in rats with polycystic ovary syndrome and its possible mechanism.Methods 48 female rats were selected to establish the model of polycystic ovary syndrome(PCOS)by intragastric administration of letrozole.The successful modeled rats were randomly divided into model group,low and high dose ASP groups and western medicine group,with 12 rats in each group.Another control group(12 rats)was set up.Low and high dose of ASP groups were given 50 and 200 mg·kg^(-1) ASP(prepared with normal saline)by gavage,the western medicine group was given 200 mg·kg^(-1) metformin solution by gavage,the control and model group were given the same amount of normal saline for 28 days.The levels of serum estradiol(E 2),testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH)and malondialdehyde(MDA),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD)were measured by enzyme linked immunosorbent assay(ELISA).The ovarian mass,volume and morphological changes of rat ovary were calculated and analyzed.The pathological changes of rat ovary in each group were observed by HE staining.The mRNA and protein expressions of Nrf2 and HO-1 in rat ovary were detected by real time quantitative polymerase chain reaction(RT-qPCR)and Western blotting.Results Compared with the model group,the levels of serum E 2,FSH,GSH-Px,SOD,Nrf2,HO-1 mRNA and protein expression were increased,and the levels of T,LH,MDA,ovarian mass and volume were decreased in the low and high dose ASP groups and western medicine group(P<0.05).Compared with the low dose ASP group,the indexes of ASP high dose group and Western medicine group changed significantly(P<0.05).Conclusion ASP can adjust the level of serum oxidative factors,reduce oxidative stress and improve ovarian cystic dilatation in PCOS rats,which may play a role by activating Nrf2/HO-1 signal pathway.