摘要
目的:观察电针肝经腧穴对卵巢储备功能下降(diminished ovarian reserve,DOR)模型大鼠性激素水平的影响,探讨电针肝经腧穴通过调控Smad2、smad3蛋白表达水平影响TGF-β1分泌治疗DOR的作用与机制。方法:将32只清洁级SD雌性大鼠随机分为空白组、模型组、肝经组、非经非穴组,每组8只。采用腹腔注射环磷酰胺(CTX)建立DOR大鼠模型。肝经组取“曲泉穴”、“太冲穴”、“急脉穴”、“蠡沟穴”电针治疗,非经非穴组取四个非经非穴点电针治疗,隔日1次,每周3次,共治疗3周。治疗结束后,观察大鼠一般情况;HE染色观察大鼠动情周期变化;酶联免疫吸附剂测定法(ELISA法)检测血清中雌二醇(E2)、促卵泡生成素(FSH)、促黄体生成素(LH)、抗缪勒激素(AMH)水平;蛋白质印迹(Western Blot)检测大鼠卵巢组织TGF-β1、Smad2、smad3蛋白表达。结果:与空白组相比,模型组大鼠体重减少,行为习惯异常,动情周期不同程度延长及紊乱,E2、AMH水平明显降低(P<0.01),FSH、LH水平升高(P<0.01),TGF-β1、Smad2、smad3蛋白表达水平显著下降(P<0.01);与模型组相比,肝经组大鼠体重、活动恢复,动情周期规律,血清E2、AMH水平升高(P<0.01),FSH、LH水平下降(P<0.01),TGF-β1、Smad2、smad3蛋白表达水平明显升高(P<0.01),非经非穴组大鼠异常表现均减轻,动情周期紊乱率降低,E2、AMH水平升高(P<0.01,P<0.05),FSH、LH含量下降(P<0.05);与非经非穴组相比,肝经组大鼠动情周期规律,E2、AMH水平升高(P<0.05),FSH、LH水平下降(P<0.05),TGF-β1、Smad2、smad3蛋白表达水平强表达且显著升高(P<0.01)。结论:电针肝经腧穴能够改善DOR大鼠卵巢储备功能,其机制可能与调控TGF-β1及Smad2、3蛋白的表达有关。
Objective:To observe the effect of electroacupuncture at acupoints of liver meridian on the level of sex hormones in rats with decreased ovarian reserve function(diminished ovarian reserve,DOR),and to explore the effect and mechanism of electroacupuncture at acupoints of liver meridian on the secretion of TGF-β1 in the treatment of DOR by regulating the expression of Smad2and Smad3protein.Methods:32clean female SD rats were randomly divided into blank group(n=8),model group(n=8),liver meridian group(n=8)and non-acupoint group(n=8).The rat model of DOR was established by intraperitoneal injection of cyclophosphamide(CTX).The liver meridian group was treated with electroacupuncture at“Ququan acupoint”,“Taichong acupoint”,“Jimai acupoint”and“Ligou acupoint”,and the non-meridian non-acupoint group was treated with electroacupuncture at non-meridian and non-acupoints,once every other day,3times a week for 3weeks.After treatment,the general condition of rats was observed,the estrous cycle of rats was observed by HE staining,the levels of estradiol(E2),follicle stimulating hormone(FSH),luteinizing hormone(LH)and anti-Mullerian hormone(AMH)in serum were detected by enzyme-linked immunosorbent assay(ELISA),and the protein expressions of TGF-β1,Smad2and Smad3in rat ovary were detected by western blot.Results:Compared with the blank group,the body weight of the model group was decreased,the behavior habit was abnormal,the estrous cycle was prolonged and disordered in varying degrees,the levels of E2and AMH were significantly decreased,the levels of FSH and LH were increased,and the protein expression levels of TGF-β1,Smad2and Smad3were significantly decreased in the model group.Compared with the model group,the body weight,activity,estrous cycle regularity,serum E2and AMH levels increased,FSH and LH levels decreased,TGF-β1,Smad2and Smad3protein expression increased significantly in the liver meridian group.The abnormal performance,estrous cycle disorder rate and estrous cycle disorder rate were alleviated in the non-acupoint group(P<0.01).The contents of FSH and LH decreased.Compared with the non-acupoint group,the estrous cycle of rats in the liver meridian group was regular,the levels of E2and AMH were increased,the levels of FSH and LH were decreased,and the protein expressions of TGF-β1,Smad2and Smad3were strongly ex-pressed and significantly increased.Conclusion:Electroacupuncture at acupoints of liver meridian can improve the ovarian reserve function of DOR rats,and its mechanism may be related to the regulation of TGF-β1,Smad2and Smad3protein expression.
作者
黄丽
杨孝芳
陈兰县
陈可欣
陈皖倩
杨志虹
刘佳敏
HUANG Li;YANG Xiaofang;CHEN Lanxian(Guizhou University of Traditional Chinese Medicine,Guiyang Guizhou 550025,China)
出处
《四川中医》
2024年第12期50-53,共4页
Journal of Sichuan of Traditional Chinese Medicine
基金
国家重点研发项目(编号:2019YFC1709104)
贵州中医药大学学术新苗(贵科合学术新苗[2023]-44号)
贵州中医药大学大学生创新创业训练计划项目(贵中医大创合字(2021)62号)。
作者简介
第一作者:黄丽,硕士研究生在读,研究方向:针灸治疗心脑血管疾病及盆底病临床及机制研究,E-mail:3509576386@qq.com;通讯作者:杨志虹,硕士,硕士生导师,高级实验师,研究方向:针灸治疗心脑血管疾病及盆底病临床及机制研究,E-mail:yangmiss1115@163.com。
