H_(2)O_(2)诱导的犊牛睾丸支持细胞氧化应激模型的建立及其效果  

Modelling oxidative stress in calf sertoli cells induced by H_(2)O_(2) and its effect analysis

作　　者：唐颖 王子铭 王皓 陈艳茹 РОТАРЬЛюбовьНиколаевна 郑鹏[1] TANG Ying;WANG Ziming;WANG Hao;CHEN Yanru;ROTAR Lubov Nikolaevna;ZHENG Peng(College of Animal Science and Technology,Northeast Agricultural University,Harbin 150030,China;Animal Husbandry and Veterinary Medicine Branch of Heilongjiang Academy of Agricultural Sciences,Qiqihar 161000,China;Saint Petersburg State Agrarian University,Saint Petersburg 196601,Russia)

机构地区：[1]东北农业大学动物科学技术学院,黑龙江哈尔滨150030 [2]黑龙江省农业科学院畜牧兽医分院,黑龙江齐齐哈尔161000 [3]圣彼得堡国立农业大学,俄罗斯圣彼得堡196601

出　　处：《黑龙江动物繁殖》2025年第1期18-23,F0003,共7页Heilongjiang journal of animal reproduction

基　　金：黑龙江省自然科学基金项目(LH2020C019)。

摘　　要：雄性家畜睾丸中的氧化应激会导致其繁殖能力下降。为了建立睾丸支持细胞的氧化应激模型,为研究睾丸的氧化应激机制奠定基础,试验使用50,100,300μmol/L过氧化氢(H_(2)O_(2))处理犊牛睾丸支持细胞,建立睾丸支持细胞氧化应激模型,以未处理细胞作对照,通过检测细胞活力和丙二醛(MDA)含量筛选最佳H_(2)O_(2)处理浓度,然后检测最佳H_(2)O_(2)处理浓度下细胞的活性氧(ROS)含量及谷胱甘肽(GSH)和超氧化物歧化酶(SOD)活性,并利用实时定量PCR分析核因子相关因子2(Nrf2)、Kelch样ECH关联蛋白1(Keap1)、血红素加氧酶-1(HO-1)、NADPH醌脱氢酶1(NQO1)基因表达水平。结果表明:与对照相比,用300μmol/L H_(2)O_(2)处理的细胞活力显著降低(P<0.05),且显著低于50,100μmol/L H_(2)O_(2)处理的细胞(P<0.05);MDA含量显著增加(P<0.05),且显著高于50,100μmol/L H_(2)O_(2)处理的细胞,说明300μmol/L H_(2)O_(2)处理浓度为最佳浓度。与对照相比,300μmol/L H_(2)O_(2)处理的细胞中ROS含量显著增加(P<0.05),GSH、SOD活性显著降低(P<0.05),Nrf2、Keap1、HO-1和NQO1基因的mRNA表达量显著降低(P<0.05)。说明300μmol/LH_(2)O_(2)能够诱导犊牛睾丸支持细胞的氧化应激模型。Oxidative stress in the testicles of male livestock can lead to reduced reproductive performance.To establish an oxidative stress model of testicular sertoli cells and lay a foundation for studying the oxidative stress mechanism in the testis,the calf testicular sertoli cells were treated with 50,100,and 300μmol/L hydrogen peroxide(H_(2)O_(2)),respectively,and untreated cells were used as controls.The optimal H_(2)O_(2) treatment concentration was screened by detecting cell viability and malondialdehyde(MDA)content,and then the reactive oxygen species(ROS),glutathione(GSH)and superoxide dismutase(SOD)activities of cells under the optimal H_(2)O_(2) treatment concentration were detected.Real-time quantitative PCR was used to analyze the expression of nuclear factor-related factor 2(Nrf2),Kelch-like ECH-associated protein 1(Keap1),heme oxygenase-1(HO-1)and NADPH quinone dehydrogenase 1(NQO1)gene expression levels.The results showed that compared with the control,the viability of cells treated with 300μmol/L H_(2)O_(2) was significantly reduced(P<0.05),and was significantly lower than that of cells treated with 50 and 100μmol/L H_(2)O_(2)(P<0.05),respectively.The MDA content from treatment of 300μmol/L H_(2)O_(2) was significantly increased(P<0.05)and was significantly higher than that of cells treated with 50 and 100μmol/L H_(2)O_(2),indicating that 300μmol/L H_(2)O_(2) was the optimal treatment concentration in this experiment.Compared with the control,the ROS content in cells treated with 300μmol/L H_(2)O_(2) was significantly increased(P<0.05),the activities of GSH and SOD were significantly decreased(P<0.05),and the mRNA expression levels of Nrf2,Keap1,HO-1,and NQO1 genes were significantly decreased(P<0.05).It could be concluded that 300μmol/L H_(2)O_(2) would be preferred for modelling oxidative stress in calf testicular sertoli cells.

关 键 词：犊牛 睾丸 支持细胞 氧化应激 活性氧(ROS) 核因子相关因子2(Nrf2)基因 

分 类 号：S823.3[农业科学—畜牧学]