首乌丸对卵巢早衰模型大鼠SIRT1/PGC-1α/Cytochrome C通路的影响  

Effect of Shouwu Pill on SIRT1/PGC-1α/Cytochrome C Pathway in Premature Ovarian Failure Model Rats

作　　者：加秀凤 张超 罗丹 汤琪 陈刚 JIA Xiufeng;ZHANG Chao;LUO Dan;TANG Qi;CHEN Gang(School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China;Wuhan Hospital of Traditional Chinese and Western Medicine,Wuhan 430022,China;Hubei Provincial Hospital of TCM,Hubei University of Chinese Medicine,Wuhan 430014,China)

机构地区：[1]湖北中医药大学基础医学院,武汉430065 [2]武汉市中西医结合医院,武汉430022 [3]湖北中医药大学湖北省中医院,武汉430061

出　　处：《中国中医基础医学杂志》2023年第3期413-419,共7页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：湖北省卫生计生委中西医结合重点科研项目[鄂卫生计生通(2017)20号];湖北省第二届医学领军人才工程培养对象暨湖北名医工作室负责人项目[鄂卫通(2019)47号];湖北省卫生健康委员会中医药科研项目青年人才项目(ZY2021Q031)。

摘　　要：目的探讨首乌丸是否通过调控线粒体介导的细胞凋亡通路抑制卵巢早衰(premature ovarian failure,POF)进展。方法60只SD雌性大鼠随机分为空白组、模型组、首乌丸4周组、首乌丸6周组、首乌丸预防组、补佳乐组。腹腔注射去氧乙烯基环己烯(deoxyvinylcyclohexene,VCD)建立POF大鼠模型,予相应药物灌胃治疗后,采用Elisa法检测各组大鼠血清性激素水平,HE染色观察卵巢组织结构变化,电镜观察卵巢超微结构变化,荧光标记TUNEL法检测卵巢细胞凋亡水平,荧光定量PCR及Western blot检测大鼠卵巢组织中沉默信息调节因子2相关酶1(silent mating type information regulation 2 homolog 1,SIRT1)、过氧化物酶增殖活化受体γ辅助活化因子1α(peroxisome proliferator-activated receptorγcoactivator1α,PGC-1α)、抑凋亡因子B淋巴细胞瘤-2(b-cell lymphoma-2,Bcl-2)、促凋亡因子细胞色素C(cytochrome C,Cyt C)及Bcl-2相关X蛋白(bcl2-associated X protein,Bax)的mRNA及蛋白表达水平。结果与空白组比较,模型组大鼠血清促卵泡激素(follicle stimulating hormone,FSH)和黄体生成素(luteinizing hormone,LH)水平显著升高(P<0.01),抗缪勒氏管激素(anti-mullerian hormone,AMH)、雌激素(estradiol,E2)水平显著降低(P<0.01);大鼠卵巢组织结构被破坏,各级卵泡减少,卵巢颗粒细胞(granulosa cells,GCs)内线粒体减少;细胞凋亡指数显著升高(P<0.01);SIRT1、PGC-1α及Bcl-2的mRNA及蛋白表达均显著下降(P<0.01),Cyt C及Bax的mRNA及蛋白表达均显著上升(P<0.01)。与模型组比较,首乌丸各治疗组血清性激素水平均显著改善(P<0.01,P<0.05);大鼠卵巢萎缩减轻,各级卵泡增多;卵巢超微结构改善,线粒体数量增加,嵴增多;细胞凋亡指数显著下降(P<0.05);SIRT1、PGC-1α及Bcl-2的mRNA及蛋白表达均显著上升(P<0.05),Cyt C及Bax的mRNA及蛋白表达均显著下降(P<0.05),其中首乌丸预防组的作用最明显。结论首乌丸可以延缓POF进展,早期预防作用更显Objective To explore whether Shouwu Pill inhibit the progression of premature ovarian failure(POF)via regulating the mitochondrial-mediated apoptosis signal pathway.Methods Sixty SPF female SD rats were randomly divided into blank group,model group,Shouwu Pill-4 weeks group,Shouwu Pill-6 weeks group,Shouwu Pill Prevention group and Progynova(estradiol valerate)group.POF was induced in rats with subcutaneously injection of deoxyvinylcyclohexene(VCD)dissolved in DMSO(160 mg/kg).After administration of corresponding drugs by gavage,serum hormone levels,ovarian pathology and cell apoptosis were detected by Elisa,HE staining,electron microscopy and fluorescent-labeled TUNEL,respectively.The signal pathway were evaluated with Real-time PCR and Western blot.Results Compared with blank group,Follicle Stimulating Hormone(FSH)and Luteinizing Hormone(LH)levels in serum of model group were significantly increased(P<0.05),and anti-Mullerian Hormone(AMH)and estradiol(E2)levels were significantly decreased(P<0.05),ovarian morphology and granulosa cells(GCs)ultrastructure was destroyed,the follicles at various developmental stages were decreased,and the mitochondria in GCs were reduced,the apoptotic cells were significantly increased,the expression of Silent mating type information regulation 2 homolog 1(SIRT1),Peroxisome proliferator-activated receptor Ycoactivator-1α(PGC-1α)and B-cell lymphoma-2(Bcl-2)mRNA and protein decreased significantly(P<0.05),the expression of mRNA and protein of Cytochrome C(Cyt C)and Bcl2-associated X protein(Bax)increased significantly(P<0.05).Compared with model group,serum sex hormone levels after administration of Shouwu Pill were significantly improved(P<0.05),ovarian morphology and GCs ultrastructure were significantly ameliorated,folliclesat various developmental stages increased,the mitochondria were significantly increased,the apoptotic degree was significantly lower(P<0.05),the expression of SIRT1,PGC-1αand Bcl-2 increased significantly(P<0.05),the expression of Cyt C and Bax decreased si

关 键 词：首乌丸 卵巢早衰 SIRT1/PGC-1α/Cyt C信号通路 线粒体 大鼠 

分 类 号：R289.9[医药卫生—方剂学]