摘要
目的探讨维生素D(vitamin D,VD)对多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠卵巢颗粒细胞(granulosa cells,GCs)凋亡的影响。方法选取3周龄SD雌性大鼠21只,采用随机数字表法分为对照组(n=6)和造模组(n=15),建立PCOS大鼠模型。造模组采用来曲唑[1mg/(kg·d),溶于0.5%羧甲纤维素钠溶液]灌胃联合高脂饲料喂养,对照组采用等量的0.5%羧甲基纤维素钠溶液灌胃,给予普通饲料喂养,将12只造模成功的大鼠(3只造模失败),采用随机数字表法分为PCOS组(n=6)和PCOS+VD组(n=6),两组灌胃同等剂量玉米油,普通饲料喂养,连续21d。比较对照组、PCOS组和PCOS+VD组大鼠的动情周期、卵巢形态学变化,以及血清25羟维生素D[25 hydroxyvitam D,25-(OH)D]、睾酮(testosterone,T)、黄体生成素(luteinizing hormone,LH)、促卵泡激素(follicle-stimulating hormone,FSH)水平。造模结束后,分离GCs进行体外培养,根据培养方式分为GCs对照组(n=6)、GCs-PCOS组(n=6)、GCs-(PCOS+VD)组(n=6)、GCs-(PCOS+VD+LY294002)组(n=6)。采用流式细胞术检测各组细胞凋亡率,透射电镜观察细胞凋亡小体,蛋白免疫印迹法检测各组细胞中Bax、Bcl-2、PI3K、p-PI3K、p-Akt、Akt、p-mTOR、mTOR蛋白表达。结果与对照组比较,PCOS组大鼠血清中T、LH水平升高,FSH、25-(OH)D水平降低,动情周期紊乱,卵巢呈多囊样改变,差异均有统计学意义(P<0.05);与PCOS组比较,PCOS+VD组大鼠血清T、LH降低,FSH及25-(OH)D水平升高,动情周期恢复正常,囊状卵泡数量减少,差异均有统计学意义(P<0.05)。与GCs对照组比较,GCs-PCOS组GCs凋亡率及Bax蛋白表达增加,Bcl-2、p-PI3K、p-AKT、p-mTOR蛋白表达减少,差异均有统计学意义(P<0.05);与GCs-PCOS组比较,GCs-(PCOS+VD)组的GCs凋亡率及Bax蛋白表达降低,Bcl-2、p-PI3K、p-AKT、p-mTOR蛋白表达增加,差异均有统计学意义(P<0.05);与GCs-(PCOS+VD)组比较,GCs-(PCOS+VD+LY294002)组的p-PI3K、p-AKT、p-mTOR蛋白表达明显降低,差异均�
Objective To investigate the influence of vitamin D(VD)on apoptosis of ovarian granulosa cells(GCs)in rats with polycystic ovary syndrome(PCOS).Methods There were 213-week-old female SD rats,which were divided into control group(n=6)and modeling group(n=15)according to random number table method to establish the PCOS rat model.The modeling group was fed with letrozole[1mg/(kg·d)dissolved in 0.5%sodium carboxymethylcellulose solution]by gavage combined with high-fat chow,while the control group was fed with an equal amount of 0.5%sodium carboxymethylcellulose solution by gavage and given normal chow.12 rats with successful modeling(3 rats with failed modeling)were divided into PCOS group(n=6)and PCOS+VD group(n=6).Both groups were fed with equal doses of corn oil by gavage and normal feed for 21 days.The rats in the control group,PCOS group and PCOS+VD group were compared in terms of their motility cycle,ovarian morphology,serum 25 hydroxyvitam D[25-(OH)D],testosterone(T),luteinizing hormone(LH),follicle-supporting hormone(FSH).After modelling,GCs were isolated and cultured in vitro and divided into GCs control group(n=6),GCs-PCOS group(n=6),GCs-(PCOS+VD)group(n=6)and GCs-(PCOS+VD+LY294002)group(n=6)according to the culture method.The apoptosis rate of each group was detected by flow cytometry,apoptotic vesicles were observed by transmission electron microscopy and Bax,Bcl-2,PI3K,p-PI3K,p-Akt,Akt,p-m TOR and m TOR protein expression were detected in each group of cells by protein immunoblotting.Results Compared with the control group,rats in the PCOS group showed increased serum T and LH levels,decreased FSH and 25-(OH)D levels,disturbed motility cycle and polycystic ovarian changes,and the differences were statistically significant(P<0.05).Compared with the PCOS group,the serum T and LH levels in the PCOS+VD group decreased,FSH and 25-(OH)D levels increased,and the motile cycle returned to normal,and the differences were statistically significant(P<0.05).Compared with the GCs control group,the GCs-PCOS group sho
作者
孙婷
马思琪
郜艳翠
刘远航
李琳
曲银娥
SUN Ting;MA Siqi;GAO Yancui;LIU Yuanhang;LI LIN;QU Yine(School of Basic Medical Sciences,North China University of Technology,Hebei Key Laboratory of Chronic Diseases,Tangshan 063210,Hebei,China)
出处
《中国现代医生》
2023年第5期68-74,共7页
China Modern Doctor
基金
河北省引进留学人员资助项目(C201860)。
