摘要
目的探讨左归丸对自然衰老小鼠卵母细胞线粒体功能的改善作用。方法将10只2月龄小鼠作为青年对照组,将30只11月龄小鼠随机分为衰老空白组、左归丸低剂量组(1.2 g·kg-1,相当于临床用量)和左归丸高剂量组(12.0 g·kg-1),每组10只。小鼠灌胃给药,每天1次,连续60 d。末次给药后,小鼠腹腔注射孕马血清促性腺激素(PMSG,5 IU),48 h后再注射人绒毛膜促性腺激素(hCG,8~10 IU)诱导。16 h后解剖并分离卵母细胞。采用生物发光法测定卵母细胞三磷酸腺苷(ATP)含量;电镜观察线粒体的数目、形态和结构;JC-1染色法检测线粒体膜电位;Western Blot法检测卵母细胞内过氧化物酶体增殖物激活受体γ辅助活化因子1α(PCG-1α)、视神经萎缩蛋白1(OPA1)和PTEN诱导蛋白激酶1(PINK1)的表达水平。结果与青年对照组比较,衰老空白组小鼠的卵母细胞ATP含量明显降低(P<0.01),线粒体数目明显减少(P<0.01),空泡增多,膜电位降低(P<0.01),PCG-1α、OPA1及PINK1蛋白表达水平明显下调(P<0.01)。与衰老空白组比较,左归丸高、低剂量组小鼠的卵母细胞ATP含量明显提高(P<0.01),线粒体结构得到改善且数目明显提高(P<0.05),线粒体膜电位明显升高(P<0.01),PCG-1α、OPA1及PINK1蛋白表达明显上调(P<0.01)。结论卵母细胞线粒体功能降低可能是小鼠卵母细胞衰老的机制之一,而左归丸能够改善自然衰老的线粒体功能,可能与其通过促进线粒体自我更新的动力过程有关。
Objective To investigate the effect of Zuogui Wan on improving mitochondrial energy metabolism and structure and the underlying mechanism in ageing mice oocytes.Methods Ten young female mice(2 months)were assigned to young control group;30 female mice were divided into three groups:11 months control aging group and1.20 g·kg-1 Zuogui Wan group,and 12.0 g·kg-1 Zuogui Wan group.After administrated with Zuogui Wan via intragastric gavage for 60 days,induction was carried out by intraperitoneal injections of pregnant mare’s serum gonadotropin(PMSG)followed by hCG 48 h later.Mice were killed and oocytes were acquired approximately 16 h after the hCG injection.The levels of adenosine triphosphate(ATP)was detected by bioluminescent assay;the mitochondria was observed using transmission electron microscope;mitochondrial membrane potential(MMP)was detected with JC-1 assay kit;peroxisome proliferator activated receptorγcoactivator-1α(PGC-lα),optic atrophy 1(OPA1)and PTEN induced putative kinase 1(PINK1)were determined by Western Blot.Results Compared with the ageing mice oocytes from young mice,the levels of ATP and MMP were decreased(P<0.01),the number and structure of the mitochondria were disordered(P<0.01),the expression of PGC-lα,OPA1 and PINK1 were evidently down-regulated in aging mice(P<0.01).It was found that Zuogui Wan could promote the levels of ATP(P<0.01).The number and structure of the mitochondria in the oocytes were improved(P<0.01),MMP were elevated(P<0.01),the expression of PGC-lα,OPA1 and PINK1 were significantly up-regulated in aging mice with Zuogui Wan high and low doses treatment.Conclusion The dysfunction of mitochondria is one of mechanisms underlying of the aging of mice oocytes;it was demonstrated that Zuogui Wan could protect the structure and function of mitochondria in oocytes of aging mice,improve the dynamics of mitochondria,these changes could be due to improving the mitochondrial dynamics of aging oocytes.
作者
赵新永
孙岩
赵鸿森
周飞
ZHAO Xinyong;SUN Yan;ZHAO Hongsen;ZHOU Fei(The Medical School,Zhengzhou University of Industrial Technology,Zhengzhou 451150 Henan,China;Department of Stomatology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052 Henan,China;Fudan University Subsidiary Pudong Hospital,Shanghai 201399,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2020年第8期924-929,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金面上项目(81573722)
郑州工业应用技术学院2018年高层次人才科研启动基金项目(2018ZGGC001)
上海市高级中西医结合人才培养项目(ZY3-PCPY-4-2006)。
