绞股蓝多糖对H_(2)O_(2)致体外成熟小鼠卵母细胞氧化损伤及凋亡的影响  

Effects of Gynostemma Pentaphyllum polysaccharide on oxidative injury and apoptosis of mouse oocytes matured in vitro induced by H_(2)O_(2)

作　　者：唐非台 刘可可 杜倩笙 唐小川 何家康[1] 王晓丽[1] TANG Feitai;LIU Keke;DU Qiansheng;TANG Xiaochuan;HE Jiakang;WANG Xiaoli(College of Animal Science and Technology,Guangxi University,Nanning 530005,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530005

出　　处：《中国兽医科学》2024年第11期1532-1539,共8页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31460644);广西自然科学基金项目(2018GXNSFAA281020)。

摘　　要：为了探究绞股蓝多糖(Gynostemma penttaphyllum polysaccharides,GPP)对H_(2)O_(2)诱导小鼠卵母细胞氧化损伤及凋亡的影响,本研究将收集的小鼠第二次减数分裂中期(metaphaseⅡ,MⅡ)卵母细胞随机分为空白对照组、H_(2)O_(2)组(在成熟培养液中添加200μmol/L H_(2)O_(2))和H_(2)O_(2)+GPP组(在含H_(2)O_(2)的成熟培养液中添加200μg/m L GPP)。利用多功能酶标仪、免疫荧光以及qRT-PCR检测技术分别对各组小鼠卵母细胞中ROS、GSH、SOD、MDA含量,凋亡相关基因Bcl-2、Bax和Caspase-9的表达进行检测。结果发现,H_(2)O_(2)+GPP组中小鼠卵母细胞SOD、GSH水平均显著高于H_(2)O_(2)组(P<0.05),ROS和MDA的含量均显著低于H_(2)O_(2)组(P<0.05),与对照组差异不显著;qRT-PCR检测发现,H_(2)O_(2)+GPP组与对照组小鼠卵母细胞中SODm RNA、GSH m RNA的表达均显著高于H_(2)O_(2)组(P<0.05),与对照组差异不显著。Annexin V-m Cherry标记后,H_(2)O_(2)+GPP组与对照组卵母细胞的早期凋亡率差异不显著,但均显著低于H_(2)O_(2)组(P<0.05);H_(2)O_(2)+GPP组小鼠卵母细胞中Bcl-2 m RNA和蛋白表达均显著高于H_(2)O_(2)组(P<0.05),Bax m RNA和蛋白表达均显著低于H_(2)O_(2)组(P<0.05),Caspase-9蛋白表达显著低于H_(2)O_(2)组(P<0.05)。结果表明,在IVM培养液中添加GPP有助于改善H_(2)O_(2)导致的小鼠卵母细胞氧化损伤,降低早期凋亡率,上调抗凋亡基因表达水平,下调促凋亡基因表达水平,增强小鼠卵母细胞抗氧化作用。In order to explore the effects of Gynostemma pentaphyllum polysaccharides(GPP)on oxidative injury and apoptosis of mouse oocytes induced by H_(2)O_(2),the metaphaseⅡ(MⅡ)oocytes of mouse were randomly divided into blank control group,H_(2)O_(2) group(with 200μmol/L H_(2)O_(2) added to the mature culture medium)and H_(2)O_(2)+GPP group(with 200μg/m L GPP added to the mature culture medium containing H_(2)O_(2)).The contents of ROS,GSH,SOD,MDA,early apoptosis of oocytes,and the gene and protein expressions of Bcl-2,Bax and Caspase-9 were detected by multifunctional enzyme scale instrument,immunofluorescence and q RT-PCR detection technology.The results showed that the SOD and GSH levels of mouse oocytes in H_(2)O_(2)+GPP group were significantly higher than those in H_(2)O_(2) group(P<0.05),and the contents of ROS and MDA were significantly lower than those in H_(2)O_(2) group(P<0.05),but there was no significant difference between the two groups.The results showed that the SOD m RNA and GSH m RNA expressions of mouse oocytes in H_(2)O_(2)+GPP group and control group were significantly higher than those in H_(2)O_(2) group(P<0.05),but there was no significant difference between the two groups.After Annexin V-m Cherry labeling,the early apoptosis rate of oocytes in H_(2)O_(2)+GPP group and control group was not significantly different,but was significantly lower than that in H_(2)O_(2)group(P<0.05).The Bcl-2 m RNA and protein expressions of mouse oocytes in H_(2)O_(2)+GPP group were significantly higher than those in H_(2)O_(2) group(P<0.05),the Bax m RNA and protein expressions were significantly lower than those in H_(2)O_(2) group(P<0.05),and the Caspase-9 protein expression was significantly lower than that in H_(2)O_(2) group(P<0.05).The results indicate that the addition of GPP in IVM culture medium can help to improve the oxidative injury of mouse oocytes caused by H_(2)O_(2),reduce the early apoptosis rate,up-regulate the expression level of anti-apoptotic genes,down-regulate the expression level of pr

关 键 词：绞股蓝多糖 H_(2)O_(2) 卵母细胞 小鼠 氧化损伤 凋亡 

分 类 号：S858.91[农业科学—临床兽医学]