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作 者:李丽 宗道宽 尹萍[1] 陈媛媛 李凯[1] 蔡靓 路璐 张武文 童国庆[1] LI Li;ZONG Dao-kuan;YIN Ping;CHEN Yuan-yuan;LI Kai;CAI Liang;LU Lu;ZHANG Wu-wen;TONG Guo-qing(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai,200120,China)
机构地区:[1]上海中医药大学附属曙光医院,上海200120
出 处:《现代生物医学进展》2020年第6期1028-1032,1056,共6页Progress in Modern Biomedicine
基 金:国家自然科学基金面上项目(81571442);国家自然科学基金面上项目(81170571);上海中医药大学预算内项目(2016YSN47);曙光医院四明基金项目。
摘 要:目的:探讨过氧化氧化还原蛋白2(PRDX2)在乙醇所致的雄性小鼠生殖损害中的作用机制。方法:以雄性昆明小鼠为研究对象,分为对照组及模型组,分别以蒸馏水和乙醇灌胃处理12周,采集血清用于激素水平测定;收集精子,一部分用于精液分析,另一部分用于Q-PCR检测PRDX2、BCL-2、BAX、Caspase3的mRNA表达,Western blot检测PRDX2、BCL-2、BAX、Caspase3和Cleaved-Caspase3的蛋白表达,免疫荧光法鉴定PRDX2的表达;统计分析研究结果。结果:与对照组比较,模型组小鼠的精子活率降低,雌激素水平升高而雄激素水平降低(P<0.05);模型组小鼠的PRDX2、BCL-2蛋白及mRNA水平较对照组均降低(P<0.05),免疫荧光显示PRDX2可在精子中表达,且模型组小鼠精子的荧光强度明显减低;在BAX及Caspase3 mRNA的表达上,模型组高于对照组(P<0.05),模型组小鼠的BAX及Cleaved-Caspase3的蛋白表达亦高于对照组(P<0.05);Pearson相关系数分析显示PRDX2与BCl-2呈正相关、与BAX、Caspase3呈负相关,差异均有统计学意义(P<0.05)。结论:乙醇造成精子PRDX2表达降低,间接诱导精子发生凋亡,影响精子的生成和发育,损害雄性小鼠的生殖功能。Objective:To investigate the mechanism of peroxiredoxin 2(PRDX2)in ethanol-induced reproductive damage in male mice.Methods:Male Kunming mice were divided into control group and model group.The control group was treated with distilled water and the model group was treated with ethanol for 12 weeks.Serum was collected for hormone level determination.Sperm was collected,one part was used for semen analysis,the other part was used for Q-PCR detecting the m RNA expressions of PRDX2,BCL-2,BAX and Caspase3,Western blot detecting the protein expressions of PRDX2,BCL-2,BAX,Caspase3 and Cleaved-Caspase3 and Immunofluorescence staining identifying the expression of PRDX2.Statistical analysis results.Results:Compared with the control group,the sperm motility and the androgen level of the model group decreased,the estrogen level increased(P<0.05).Compared with the control group,the PRDX2 and BCL-2 protein and m RNA levels of the model group were significantly decreased(P<0.05).Immunofluorescence showed that PRDX2 could be expressed in sperm,and the fluorescence intensity of sperm in model group was significantly reduced.BAX and Caspase3 m RNA of the model group were significantly higher than that of the control group(P<0.05).The BAX and Cleaved-Caspase3 protein levels of the model group were significantly increased(P<0.05).Pearson correlation coefficient analysis showed that PRDX2 was positively correlated with BCl-2 and negatively correlated with BAX and Caspase3(P<0.05).Conclusion:Ethanol causes the decrease of sperm PRDX2 expression,indirectly induces sperm apoptosis,affects sperm production and development,and impairs the reproductive function of male mice.
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