基于线粒体乙酰化酶3-缺氧诱导因子1α通路探讨毓麟珠改善自然衰老大鼠卵巢氧化应激的作用机制  

To explore the mechanism of Yulinzhu in improving ovarian oxidative stress in natural aging rats based on Sirt3-HIF1a pathway

作　　者：阮鑫 魏茂林 董晓英[1] RUAN Xin;WEI Maolin;DONG Xiaoying(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院,北京100069

出　　处：《环球中医药》2024年第1期19-24,共6页Global Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(81503607);北京市自然科学基金面上项目(7212162);北京中医药薪火传承“3+3”工程项目(2012-SZ-C-41)

摘　　要：目的基于线粒体乙酰化酶3(mitochondrial acetylase 3,Sirt3)-缺氧诱导因子1α(hypoxia-inducible factor 1-alpha,HIF1α)通路探讨毓麟珠改善自然衰老大鼠卵巢氧化应激的作用机制。方法12只雌性SD老年大鼠,随机分为衰老未灌胃组与衰老灌胃组,每组6只,6只雌性正常育龄期SD大鼠作为空白对照组,衰老灌胃组予毓麟珠中剂量10.08 g/kg,衰老未灌胃组予等量生理盐水,空白对照组正常饲养,连续灌胃6周(休1天/周)。酶联免疫法测定大鼠血清激素雌二醇(estradiol,E 2)、促卵泡激素(follicle-stimulating hormone,FSH)、抗苗勒管激素(anti-mullerian hormone,AMH)水平;苏木精-伊红(hematoxylin eosin stain,HE)染色观察卵巢形态学变化;化学发光法测定卵巢组织中超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathion peroxidase,GSH-Px)、丙二醛(malondialdehyde,MDA)含量;化学发光法测定卵母细胞线粒体ATP含量;蛋白免疫印记法检测卵巢组织Sirt3、HIF1α蛋白表达;实时荧光定量PCR检测卵巢组织Sirt3、HIF1αmRNA表达。结果与衰老未灌胃组相比较,衰老灌胃组卵泡发育改善,表现为颗粒细胞层排列整齐、卵母细胞形态较规则;E 2、AMH水平升高,FSH水平降低(P<0.01);卵巢组织中SOD、GSH水平升高,MDA水平降低(P<0.01);ATP水平升高(P<0.05);Sirt3蛋白及基因表达均上调,HIF1α蛋白及基因表达均下调(P<0.05)。结论毓麟珠通过调控Sirt3、HIF1α的蛋白及基因表达,改善卵巢组织抗氧化能力进而达到延缓卵巢衰老的目的。Objective To investigate the effect of Yulinzhu on ovarian oxidative stress in aging rats and its mechanism based on Sirt3-HIF1a pathway.Methods Twelve female SD rats were randomly divided into aging without intragastric administration group and aging with intragastric administration group,with 6 rats in each group.Six female SD rats of normal reproductive age were used as the blank control group.The levels of serum hormone E 2,FSH and AMH were measured by enzyme-linked immunosorbent assay.HE staining was used to observe ovarian morphological changes.The contents of superoxide dismutase(SOD),glutathion peroxidase(GSH-Px)and malondialdehyde(MDA)in ovarian tissue were measured by chemiluminescence.Western blot was used to detect the protein expression of Sirt3 and HIF1αin ovarian tissue.The expressions of Sirt3 and HIF1αmRNA in ovarian tissues were detected by PCR.Results Compared with the aging without intragastric administration group,the follicular development was improved in the aging with intragastric administration group,which was characterized by orderly arrangement of granulosa cell layers and regular oocyte morphology;the levels of E2 and AMH were increased,and the level of FSH was decreased(P<0.01);the levels of SOD and GSH in ovarian tissue were increased,and the level of MDA was decreased(P<0.01);the level of ATP expression was increased(P<0.05);the expression of Sirt3 protein and gene was up-regulated,and the expression of HIF1αprotein and gene was down-regulated(P<0.05).Conclusion Yulinzhu can delay ovarian aging by regulating the protein and gene expressions of Sirt3 and HIF1αto improve the antioxidant capacity of ovarian tissue.

关 键 词：毓麟珠 线粒体乙酰化酶3 缺氧诱导因子1Α 卵巢衰老 氧化应激 

分 类 号：R285.5[医药卫生—中药学]