暖巢煲调控线粒体功能改善早发性卵巢功能不全的实验研究  

Experimental study of regulating mitochondrial function by Ovary-Warming Pot to improve early premature ovarian insufficiency

作　　者：周虹 李兰 刘文娥[2] 陈镇[2] 邓桂明[2] 何阳 肖小芹[1,3] ZHOU Hong;LI Lan;LIU Wen'e;CHEN Zhen;DENG Guiming;HE Yang;XIAO Xiaoqin(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;The First Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410007,China;Hunan University of Technology and Business,Changsha,Hunan 410205,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007 [3]湖南工商大学,湖南长沙410205

出　　处：《湖南中医药大学学报》2023年第6期1014-1020,共7页Journal of Hunan University of Chinese Medicine

基　　金：湖南省自然科学基金面上项目(2020JJ4469);湖南省教育厅科学研究重点项目(21A0225);湖南省中医药管理局科研计划重点项目(C2022008)。

摘　　要：目的探讨暖巢煲调控卵巢线粒体功能治疗化疗性早发性卵巢功能不全(premature ovarian insufficiency,POI)损伤小鼠,为其临床应用提供科学依据。方法从40只8周龄雌性C57BL/6小鼠中抽取8只为空白组,其余构建POI模型。将成功建模的32只小鼠随机分为模型组、暖巢煲低剂量组、暖巢煲中剂量组、暖巢煲高剂量组,每组8只。模型组、空白组予0.01 mL/g生理盐水灌胃。观察小鼠的一般状况、体质量和动情周期变化,给药3周后,称取各组小鼠卵巢湿质量,计算卵巢指数。ELISA法检测各组小鼠血清抗米勒管激素(anti-Müllerian hormone,AMH)水平,HE染色观察卵巢组织形态学变化,透射电镜观察线粒体的形态和结构,Western blot检测视神经萎缩蛋白1(optic atrophy 1,OPA1)和PTEN诱导假定激酶1(PTEN-induced putative kinase 1,PINK1)的表达水平。结果与空白组比较,模型组进食量减少、毛色枯燥;体质量增长率下降(P<0.05);卵巢指数明显下降(P<0.01);动情周期紊乱;血清AMH下降(P<0.05);各级生长卵泡减少、闭锁卵泡明显增加;线粒体结构破坏严重、空洞化明显;OPA1及PINK1蛋白表达水平明显下调(P<0.01)。与模型组比较,暖巢煲高、中、低剂量组体质量增长率上升(P<0.05);暖巢煲中、低剂量组卵巢指数增加;动情周期逐渐规律;暖巢煲中、高剂量组血清AMH上升(P<0.05),小鼠的卵母细胞线粒体结构明显改善,OPA1及PINK1蛋白表达明显上调(P<0.01)。结论线粒体功能障碍可能是小鼠卵巢功能不全的机制之一,而暖巢煲能够调节卵巢线粒体功能,改善早发性卵巢不全,可能与其通过促进线粒体自我更新的动力过程有关。Objective To discuss the clinical efficacy of Ovary-Warming Pot(OWP)on premature ovarian insufficiency(POI)mice by regulating ovarian mitochondrial function and to provide a scientific basis for its clinical application.Methods Eight of 408-week-old female C57BL/6 were selected as blank group,and the rest were used to establish the POI models.Then 32 successfully modeled mice were randomly divided into model group,low-,medium-and high-dose OWP groups,with 8 mice in each group.The model group and blank group were given 0.01mL/g normal saline by gastric lavage.The general condition,body weight,and estrous cycle of mice were observed.After 3 weeks of administration,the wet ovarian weight of mice in each group was weighed to calculate the ovarian index.The serum anti-Müllerian hormone(AMH)level was determined by enzyme-linked immunosorbent assay(ELISA),the morphological changes of ovarian tissue were observed by hematoxylin-eosin(HE)staining,and the morphology and structure of mitochondria were observed by transmission electron microscopy.Western blot was used to determine the expression levels of optic atrophy 1(OPA1)and PTEN-induced protein kinase 1(PINK1).Results Compared with blank group,the mice in model group had less food intake and dull hair color;their growth rate of body mass was lower(P<0.05);the ovarian index was significantly lower(P<0.01);the estrous cycle was disordered;the serum AMH was lower(P<0.05);the growth follicles at all levels decreased,while atresia follicles increased significantly;the mitochondrial structure was seriously damaged,with obvious cavitation;the expression levels of OPA1 and PINK1 were significantly down-regulated(P<0.01).Compared with model group,the growth rates of body mass in high-,medium-and low-dose OWP groups were higher(P<0.05);the ovarian indexes in medium-and low-dose OWP groups were higher,and the estrus cycle became regular gradually;the serum AMH in medium-and high-dose OWP groups was significantly higher(P<0.05),the mitochondrial structure of mice oocytes was sign

关 键 词：早发性卵巢功能不全 暖巢煲 线粒体 视神经萎缩蛋白1 PTEN诱导假定激酶1 抗米勒管激素 

分 类 号：R271.9[医药卫生—中西医结合]