[目的]探究绞股蓝多糖(Gynostemma penttaphyllum polysaccharides，GPP)对过氧化氢(H2O2)损伤小鼠卵母细胞及线粒体的保护作用。[方法]用H2O2构建小鼠卵母细胞体外成熟氧化应激模型，并对GPP添加浓度进行筛选。将收集的小鼠MⅡ期卵母细胞随机分为空白对照组、H2O2组(在成熟培养液中添加H2O2)和H2O2+GPP组(在含H2O2的成熟培养液中添加GPP)。体外受精后统计卵母细胞的卵裂率、4-细胞率、8-细胞率、桑椹胚率和囊胚率;利用线粒体红色荧光探针(Mito Tracker Red CMXRos)检测小鼠卵母细胞线粒体分布及含量，采用Jc-1线粒体膜电位检测试剂盒检测线粒体膜电位，利用ATP检测试剂盒检测线粒体ATP含量，通过免疫荧光染色观察小鼠卵母细胞纺锤体形态和染色体分布。[结果]H2O2+GPP组卵母细胞的卵裂率、4-细胞率、8-细胞率、桑椹胚率、囊胚率、线粒体含量、线粒体膜电位和ATP含量均显著高于H2O2组(P＜0。05)，纺锤体异常率和染色体异常率均显著低于H2O2组(P＜0。05)，与对照组间差异均不显著(P＞0。05)。[结论]GPP可提高H2O2损伤小鼠卵母细胞体外受精的卵裂率和囊胚率，增加卵母细胞内线粒体含量、线粒体正常分布、线粒体膜电位、ATP含量，降低纺锤体形态异常、染色体形态异常和排列紊乱，从而减少线粒体的功能障碍。
Protection of Gynostemma pentaphyllum Polysaccharides on Mouse Oocytes and Mitochondria Damaged by H2O2
[Objective]This study was aimed to investigate the protection of Gynostemma penttaphyllum polysaccharides(GPP)on mouse oocytes and mitochondria damaged by hydrogen peroxide(H2O2).[Method]Mouse oocyte maturation oxidative stress model was constructed with H2O2,and GPP concentration was selected.Then oocytes at M Ⅱ stage were randomly divided into blank control group,H2O2 group(H2O2 was added in mature medium)and H2O2+GPP group(GPP was added in mature medium containing H2O2).The cleavage rate,4-cell rate,8-cell rate,morula rate and blastocyst rate of oocytes were measured after in vitro fertilization(IVF).Mito Tracker Red CMXRos was used to detect mitochondrial distribution and content,JC-1 mitochondrial membrane potential detection kit was used to detect mitochondrial membrane potential,the ATP detection kit was used to detect mitochondrial ATP content,and the morphology and chromosome distribution of mouse oocytes were observed by immunofluorescence staining.[Result]The cleavage rate,4-cell rate,8-cell rate,morula rate and blastocyst rate,the mitochondrial content,mitochondrial membrane potential and ATP contents of oocytes in H2O2+GPP group were significantly higher than those in H2O2 group(P＜0.05),and the rates of spindle abnormality and chromosome abnormality were significantly lower than those in H2O2 group(P＜0.05),but there was no significant difference between H2O2+GPP and control groups(P＞0.05).[Conclusion]GPP could improve the cleavage rate and blastocyst rate of H2O2-damaged mouse oocytes IVF,as well as the mitochondrial content,mitochondrial normal distribution,mitochondrial membrane potential and ATP content in oocytes,and reduce the spindle morphological abnormalities,chromosome morphological abnormalities and disordered arrangement,reducing mitochondrial dysfunction.
万方数据
