摘要
目的:探讨归肾丸对卵巢储备功能减退(DOR)小鼠卵巢功能和自噬的影响。方法:将C57BL/6小鼠随机分为5组:正常组、模型组、归肾丸低、中、高剂量组。除正常组外,各组小鼠采用腹腔注射环磷酰胺(CTX)法建立DOR小鼠模型并给予相应药物。ELISA法检测血清抗苗勒管激素(AMH)、促卵泡生成素(FSH)、雌二醇(E_(2))、促黄体生成素(LH)水平;HE染色观察卵巢组织形态;Western blot、qRT-PCR检测卵巢组织微管相关蛋白1轻链3(LC3)、Beclin 1、p62蛋白和mRNA水平。结果:与正常组相比,模型组小鼠血清FSH、LH水平升高(P<0.05),AMH、E_(2)水平降低(P<0.05);卵巢中生长卵泡和原始卵泡的数量减少,闭锁卵泡增加;卵巢组织LC3和Beclin 1蛋白和mRNA水平增高,p62蛋白和mRNA水平降低。经归肾丸干预后,DOR小鼠血清中FSH和LH激素水平降低(P<0.05),AMH和E_(2)激素水平增高(P<0.05);卵巢形态改善,原始卵泡和生长卵泡增多,闭锁卵泡减少;卵巢组织中LC3和Beclin 1的蛋白及mRNA水平降低,p62蛋白及mRNA水平升高。结论:归肾丸可以明显改善DOR小鼠的卵巢功能,可能通过抑制卵巢组织自噬发挥作用。
Objective:to investigate the effect of Guishen Wan on ovarian function and autophagy in mice with diminished ovarian reserve(DOR).Methods:C57BL/6 mice were randomly divided into 5 groups:normal group,model group,low,medium,and high dose groups of Guishen Wan.Except the normal group,the DOR mouse models were established by intraperitoneal injection of Cyclophosphamide(CTX) and the corresponding drugs were given to the mice in each group.ELISA was used to detect anti-mullerian hormone(AMH),follicle stimulating hormone(FSH),estradiol(E_2) and luteinizing hormone(LH) in serum,HE staining to observe the morphology of ovarian tissue,Western blot and qRT-PCR to detect the levels of Microtubule-associated protein 1 light chain 3(LC3),Beclin 1,p62 protein and mRNA in ovarian tissue.Results:Compared with the normal group,FSH and LH in the model group increased(P<0.05),while AMH and E_(2) decreased(P<0.05);The number of growing follicles and primordial follicles in the ovaries decreased,while atresia follicles increased;LC3 and Beclin 1 protein and mRNA in ovarian tissue increased,while p62 protein and mRNA decreased.After the intervention with Guishen Wan,the levels of FSH and LH in the serum of DOR mice decreased(P<0.05),while the levels of AMH and E_(2) increased(P<0.05);ovarian morphology was improved,primordial and growing follicles increased,and atresia follicles decreased;LC3,Beclin 1 protein and mRNA levels decreased,while p62 protein and mRNA levels increased.Conclusion:Guishen Wan can significantly improve ovarian function in DOR mice,possibly by inhibiting ovarian tissue autophagy.
作者
朱文渊
杜彦芳
ZHU Wen-yuan;DU Yan-fang(School of Public Health,Hebei Medical University,Shijiazhuang 050017)
出处
《河北中医药学报》
2023年第4期11-15,共5页
Journal of Hebei Traditional Chinese Medicine and Pharmacology
关键词
卵巢储备功能减退
归肾丸
自噬
月经后期
闭经
滑胎
不孕症
DOR
Guishen Wan
autophagy
late menstrual period
amenorrhea
habitual abortion
infertility
