基因沉默砷中毒大鼠附睾血管内皮细胞生长因子对血管内皮细胞生长因子受体2及精子凋亡的影响  

Effects of endothelial cell growth factor on endothelial cell growth factor receptor 2 and sperm apoptosis in epididymal rats with gene-silencing arsenic poisoning

作　　者：戴研平 高晓勤[2] 罗羿轩[3] 秦海霞 DAI Yan-ping;GAO Xiao-qin;LUO Yi-xuan;QIN Hai-xia(School of Medicine,Yueyang Vocational and Technical College,Yueyang,Hunan 414000,China;不详)

机构地区：[1]岳阳职业技术学院医学院,湖南岳阳414000 [2]遵义医药高等专科学校基础医学院 [3]岳阳职业技术学院基础医学部 [4]乌兰察布医学高等专科学校组胚教研室

出　　处：《环境与健康杂志》2024年第9期753-758,F0002,共7页Journal of Environment and Health

基　　金：贵州省科学技术基金(黔科合人才团队[2014]4005号,黔科合LH字[2015]7568号)。

摘　　要：目的探讨基因沉默砷中毒大鼠附睾血管内皮细胞生长因子对血管内皮细胞生长因子受体2及精子凋亡的影响。方法将160只健康18周龄清洁级雄性SD大鼠随机分为对照(蒸馏水)组及2.4、12、60 mg/L亚砷酸钠染毒组,每组40只。采用自由饮用方式连续染毒6个月。将2.4 mg/L亚砷酸钠染毒组雄性SD大鼠进行慢病毒干扰,即分别为对照组、NC-shRNA阴性感染组、VEGF-shRNA-1病毒感染组、VEGF-shRNA-2病毒感染组、VEGF-shRNA-3病毒感染组,在每侧附睾分别注射含有不同慢病毒载体的病毒液100μl,每组6只。分别以Western blot及q-RT-PCR法检测大鼠附睾VEGF,VEGFR2的蛋白及mRNA的表达,采用流式细胞术检测精子凋亡率。结果慢病毒感染慢性砷中毒正常组大鼠附睾后VEGF、VEGFR2的mRNA及蛋白表达量均低于阴性感染组和对照组,差异有统计学意义(P<0.05),其中以VEGFshRNA-3质粒组干扰效果更为有效。与对照组和阴性感染组相比,VEGF-shRNA-3质粒干扰组精子凋亡率升高,差异有统计学意义(P<0.05)。结论附睾VEGF基因沉默可能通过抑制VEGFR2的表达,促进精子凋亡。Objective To understand the effects of vascular endothelial growth factor(VEGF)on vascular endothelial growth factor receptor 2 and sperm apoptosis in rats with gene-silencing arsenic poisoning.Methods One hundred and sixty male SD rats aged 18-weeks were randomly divided into control(distilled water)group and 2.4,12,60 mg/L sodium arsenite exposed groups,with 40 rats in each group.The rats were treated with arsenic through drinking water for 6 consecutive months.Male SD rats exposed to 2.4 mg/L sodium arsenic were subjected to lentiviral interference,namely control group,NC-shRNA negative infection group,VEGF-shRNA-1 infection group,VEGF-shRNA-2 infection group,VEGF-shRNA-3 infection group,respectively.Each epididymis was injected with 100μl virus containing different lentiviral vectors,6 in each group.The protein and mRNA expression levels of VEGF,VEGFR2 in the epididymis of rats were then detected by Western blot and quantitative real-time PCR(q-RT-PCR).The apoptosis rate of sperms was determined by using flow cytometry(FCM).Results Compared with the negative infection group and the normal control group,the mRNA and protein expression levels of VEGF and VEGFR2 significantly reduced following epididymal infection(P<0.05).Furthermore,the interference effect was stronger in the VEGFshRNA-3 group.Compared with the control group and the negative infection group,the apoptosis rate of sperms was significantly higher(P<0.05).Conclusion Epididymal VEGF gene silencing may promote sperm apoptosis by inhibiting VEGFR2 expression.

关 键 词：VEGF RNA干扰 砷中毒 精子凋亡 

分 类 号：R994.6[医药卫生—毒理学]