高雄激素血症对PCOS小鼠卵巢中非钠依赖性葡萄糖转运蛋白的影响  

Effect of hyperandrogenemia on sodium-independent glucose transporters in ovaries of PCOS mice

作　　者：陈田 孙镠佳 项雨 丁慧敏 葛红山 CHEN Tian;SUN Liu-jia;XIANG Yu;DING Hui-min;GE Hong-shan(Nanjing University of Chinese Medicine,Nanjing 210023;Taizhou People’s Hospital,Taizhou 225300)

机构地区：[1]南京中医药大学,南京210023 [2]泰州市人民医院,泰州225300

出　　处：《生殖医学杂志》2022年第6期805-812,共8页Journal of Reproductive Medicine

基　　金：国家自然科学基金(81771586)。

摘　　要：目的探讨多囊卵巢综合征(PCOS)中高雄激素血症(HA)对卵巢组织中非钠依赖性葡萄糖转运蛋白表达的影响。方法选用3周龄ICR小鼠为研究对象,随机分为对照组(8只)和脱氢表雄酮(DHEA)组(16只)。DHEA组小鼠连续21 d每日颈部皮下注射DHEA(60 mg/kg)建模,对照组注射含95%乙醇的芝麻油。分别评估两组小鼠的体重变化、血清睾酮水平及腹腔内注射糖耐量实验(IPGTT)情况。采用HE染色法观察小鼠卵巢形态学变化;运用蛋白免疫印迹(Western Blot)实验,观察非钠依赖性葡萄糖转运蛋白(GLUTs)在小鼠卵巢组织中的表达情况。同时,使用不同浓度睾酮(0、1、10μmol/L)处理人卵巢颗粒细胞(KGN细胞),采用Western Blot法检测不同睾酮浓度下KGN细胞中GLUTs蛋白的表达情况。结果与对照组相比,DHEA组小鼠体重增重率、血清睾酮水平显著增加(P<0.05),HE染色示DHEA组卵巢呈多囊样改变,提示PCOS小鼠建模成功。DHEA组小鼠空腹血糖水平及IPGTT中30、60和120 min血糖水平显著高于对照组(P<0.01)。Western Blot实验结果显示,DHEA组小鼠卵巢组织中GLUT3、4、8的表达水平显著低于对照组(P<0.05),而GLUT1和GLUT12在对照组和DHEA组中的蛋白表达水平变化不大(P>0.05)。10μmol/L睾酮处理会显著降低KGN细胞中GLUT1、GLUT4、GLUT8的表达(P<0.05),其中GLUT8对睾酮最为敏感,1μmol/L睾酮就可引起GLUT8显著降低(P<0.01);而GLUT3、GLUT12在不同浓度睾酮组中表达水平变化不大(P>0.05)。结论DHEA建模成功的PCOS小鼠卵巢中多个GLUTs表达发生了改变;HA状态会显著影响小鼠卵巢和KGN细胞中GLUTs的表达模式,提示HA对GLUTs蛋白的影响可能是其引起PCOS糖代谢异常的重要机制。Objective:To explore the effect of hyperandrogenemia(HA)on sodium-independent glucose transporters protein(GLUTs)in ovarian tissues of mice with polycystic ovary syndrome(PCOS).Methods:Three-week-old ICR mice were collected as research objects and randomly divided into control group(n=8)and dehydroepiandrosterone(DHEA)group(n=16).Mice in the DHEA group were injected with DHEA(60 mg/kg)subcutaneously on the neck for 21 days for modeling,and the mice in control group were injected with ethanol-containing sesame oil.The mouse weight,serum testosterone level and intraperitoneal glucose tolerance test(IPGTT)were evaluated in the mice of the two groups.The changes of ovarian tissue structure were observed by hematoxylin-eosin staining(HE).Western blot(WB)was used to observe the expression of GLUTs in ovarian tissue.Besides,human ovarian granulosa cells(KGN cells)were incubated with different concentrations of testosterone(0,1,10μmol/L)in vitro,and the expression of GLUTs protein in the KGN cells was evaluated by WB.Results:Compared with the control group,the weight and serum testosterone levels in the DHEA group were significantly increased(P<0.05).HE staining showed that the ovaries in DHEA group appeared polycystic changes,which suggested PCOS mouse modeling is successful.The fasting blood glucose and the blood glucose at 30,60,120 min.during IPGTT of the DHEA group were significantly higher than that of the control group(P<0.01).Western blot results showed that the expression of GLUT 3,4,8 were significantly downregulated in the PCOS model compared with the control group(P<0.05),while GLUT1 and GLUT12 had no significant change(P>0.05).Concentration 10μmol/L testosterone treatment significantly decreased the expressions of GLUT 1,4 and 8 in KGN cells(P<0.05).Among them,GLUT8 was the most sensitive to testosterone,1μmol/L testosterone could significantly reduce GLUT8 expression(P<0.01).However,GLUT3 and GLUT12 did not significantly change among the different concentrations of testosterone groups(P>0.05).Conclusions

关 键 词：多囊卵巢综合征 高雄激素 葡萄糖转运蛋白 糖代谢异常 

分 类 号：R711.75[医药卫生—妇产科学]