痰湿型PCOS大鼠卵巢颗粒细胞线粒体功能及Nox1/p-Akt/Glut4信号通路变化的研究  

Effects of Mitochondrial Function and N ox 1/p-Akt/GLUT4 Signaling Pathway in Granulosa Cells of Rat with Phlegm-dampness PCOS

作　　者：丛培玮[1] 张丽娜[1] 陈文娜[2] 尚冰[3] 吴兆利[4] CONG Pei-wei;ZHANG Li-na;CHEN Wen-na;SHANG Bing;WU Zhao-li(Teaching Experiment Center,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;School of Medical Laboratory,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Institute of Medical History and Literature,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;College of Acupuncture and Massage,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学教学实验中心,沈阳110847 [2]辽宁中医药大学医学检验学院,沈阳110847 [3]辽宁中医药大学医史文献研究院,沈阳110847 [4]辽宁中医药大学针灸推拿学院,沈阳110847

出　　处：《中国中医基础医学杂志》2021年第11期1749-1753,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目(82104735)-基于“痰湿不孕”探讨二陈汤调控铁死-能量代谢重编程改善PCOS卵泡发育障碍的机制研究;国家自然科学基金资助项目(81874372)-基于泡沫细胞源性外泌体microRNA介导的细胞间通讯探讨冠心病痰瘀互结证的发病机制及痰瘀同治法的干预作用;辽宁省自然科学基金资助项目(2019-2D-0953)-基于卵巢局部胰岛素抵抗探讨线粒体功能障碍对PCOS痰瘀互结证的影响及机制研究;辽宁省教育厅科学技术研究项目(L202038)-基于肠微生物-胆汁酸-IL-22信号轴探讨固本疏络法改善肥胖型PCOS-IR的机制研究。

摘　　要：目的:观察痰湿型多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠卵巢颗粒细胞(granulosa cells,GCs)线粒体功能及烟酰胺腺嘌呤二核苷酸磷酸氧化酶1(NADPH oxidase1,Nox1)/磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)/葡萄糖转运蛋白4(glucose transporter protein 4,Glut4)信号通路的作用,探讨“痰湿不孕”的科学内涵。方法:选取动情周期规律的SD大鼠22只,运用高脂+来曲唑灌胃法复制痰湿型PCOS大鼠并随机分为对照组10只和模型组12只。造模结束后选取动情周期延长的大鼠为模型组10只提取卵巢GCs,透射电镜观察GCs中线粒体的形态学变化,比色法检测GCs中腺嘌呤核苷三磷酸(adenine triphosphate,ATP)含量,化学发光法检测GCs中活性氧(reactive oxygen species,ROS)水平,Western blot法检测GCs中Nox1、p-Akt和Glut4蛋白表达水平。结果:模型组线粒体数量降低出现肿胀、嵴减少、空泡化改变,与对照组比较模型组GCs中ATP含量降低,ROS水平升高(P<0.01)。Nox1蛋白表达增高,p-Akt和Glut4蛋白表达降低(P<0.05,P<0.01)。结论:GCs线粒体功能下降和Nox1/p-Akt/Glut4信号通路异常是痰湿型PCOS卵泡发育异常与闭锁的可能机制。Objective:To investigate the mitochondrial function and Nox1/p-Akt/Glut4 signaling pathway in granulocyte cells(GCs)of rats with phlegm-dampness PCOS,and explore the scientific connotation of"phlegm-dampness causes infertility".Methods:Twenty-two SD rats with regular estrous period were selected.Rats were randomly divided into the control group(n=10)and model group(n=12),the models were established by high fat and letrozole intragastric.After modeling,rats with extended estrous period were selected as the model group(n=10).GCs were extracted.Transmission electron microscopy was used to observe mitochondria morphological changes of GCs;and ATP were measured by colorimetric;ROS in GCs were measured by chemiluminescence;the expression of Nox1,Akt and Glut4 in GCs were detected by Western blot.Results:The numbers of mitochondria in model group were decreased,appeard swollen,and the cristae were decreased,even vacuolated.Compared with the control group,ATP were decreased and ROS were increased in model group(P<0.01),the expression of Nox1 were increased significantly,p-Akt and Glut4 protein were decreased(P<0.05,P<0.01).Conclusion:Mitochondrial function decreased and Nox1/p-Akt/Glut4 signaling pathway abnormality of GCs were the possible mechanisms of the abnormal follicular development and atresia of phlegm-dampness PCOS.

关 键 词：痰湿证 多囊卵巢综合征 颗粒细胞 线粒体 Nox1/p-Akt/Glut4信号通路 

分 类 号：R285.5[医药卫生—中药学]