董瑞娜;刘殊;张敏;戴红;徐肖倩;

• 1:内蒙古医科大学
• 2:同煤集团职业病防治院
• 3:包头医学院

摘要(Abstract)：

邻苯二甲酸(2-乙基己基)酯(di(2-ethylhexyl)phthalate,DEHP)因在环境中的广泛分布而受到普遍关注。为探究DEHP对雄性生殖细胞的毒作用机制,将小鼠精原细胞(GC-1spd)分别暴露于终浓度为0(对照)、10、100、1 000 mg·L^(-1)DEHP的培养基中培养24 h,采用噻唑兰(MTT)法检测细胞相对活力,流式细胞仪检测细胞凋亡情况,实时荧光定量PCR法检测Bax和Bcl-2m RNA的表达情况及Western blot技术检测Bax和Bcl-2蛋白的表达水平。结果显示:在检测细胞相对活力时,1 000 mg·L(-1)DEHP的培养基中培养24 h,采用噻唑兰(MTT)法检测细胞相对活力,流式细胞仪检测细胞凋亡情况,实时荧光定量PCR法检测Bax和Bcl-2m RNA的表达情况及Western blot技术检测Bax和Bcl-2蛋白的表达水平。结果显示:在检测细胞相对活力时,1 000 mg·L^(-1)剂量组明显低于其他各剂量组,100 mg·L(-1)剂量组明显低于其他各剂量组,100 mg·L^(-1)剂量组低于对照组和10 mg·L(-1)剂量组低于对照组和10 mg·L^(-1)剂量组(P<0.05);在检测细胞凋亡时,1 000 mg·L(-1)剂量组(P<0.05);在检测细胞凋亡时,1 000 mg·L^(-1)剂量组早期凋亡率高于对照组(P<0.05);晚期凋亡率1 000 mg·L(-1)剂量组早期凋亡率高于对照组(P<0.05);晚期凋亡率1 000 mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组均明显高于对照组,1 000 mg·L(-1)剂量组均明显高于对照组,1 000 mg·L^(-1)剂量组高于10 mg·L(-1)剂量组高于10 mg·L^(-1)剂量组(P<0.05);总凋亡率呈现明显上升趋势,其中1 000 mg·L(-1)剂量组(P<0.05);总凋亡率呈现明显上升趋势,其中1 000 mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组明显高于对照组,1 000mg·L(-1)剂量组明显高于对照组,1 000mg·L^(-1)剂量组还明显高于10 mg·L(-1)剂量组还明显高于10 mg·L^(-1)剂量组(P<0.05)。实时荧光定量PCR检测结果显示:Bax m RNA表达水平1 000 mg·L(-1)剂量组(P<0.05)。实时荧光定量PCR检测结果显示:Bax m RNA表达水平1 000 mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组明显高于对照组,1 000 mg·L(-1)剂量组明显高于对照组,1 000 mg·L^(-1)剂量组与10 mg·L(-1)剂量组与10 mg·L^(-1)剂量组比较差异具有统计学意义(P<0.05);Bcl-2 m RNA表达水平1 000 mg·L(-1)剂量组比较差异具有统计学意义(P<0.05);Bcl-2 m RNA表达水平1 000 mg·L^(-1)剂量组低于10 mg·L(-1)剂量组低于10 mg·L^(-1)剂量组和对照组(P<0.05)。此外,Bax和Bcl-2 m RNA比值1 000 mg·L(-1)剂量组和对照组(P<0.05)。此外,Bax和Bcl-2 m RNA比值1 000 mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组均明显高于对照组,1 000 mg·L(-1)剂量组均明显高于对照组,1 000 mg·L^(-1)剂量组高于10 mg·L(-1)剂量组高于10 mg·L^(-1)剂量组(P<0.05)。Western blot结果显示,Bax蛋白表达水平1 000 mg·L(-1)剂量组(P<0.05)。Western blot结果显示,Bax蛋白表达水平1 000 mg·L^(-1)剂量组高于对照组(P<0.05);Bcl-2蛋白表达水平各剂量组与对照组相比差异均具有统计学意义,且1 000mg·L(-1)剂量组高于对照组(P<0.05);Bcl-2蛋白表达水平各剂量组与对照组相比差异均具有统计学意义,且1 000mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组明显低于10 mg·L(-1)剂量组明显低于10 mg·L^(-1)剂量组和对照组(P<0.05);此外,Bax/Bcl-2两蛋白比值1 000 mg·L(-1)剂量组和对照组(P<0.05);此外,Bax/Bcl-2两蛋白比值1 000 mg·L^(-1)和100 mg·L(-1)和100 mg·L^(-1)剂量组明显高于对照组,而1 000 mg·L(-1)剂量组明显高于对照组,而1 000 mg·L^(-1)剂量组高于10 mg·L(-1)剂量组高于10 mg·L^(-1)剂量组(P<0.05)。结果表明:DEHP可诱导小鼠精原细胞相对活力下降,通过抑制Bcl-2 m RNA和Bcl-2蛋白表达及促进Bax m RNA和Bax蛋白表达实现诱导精原细胞凋亡。

关键词(KeyWords)： DEHP;小鼠精原细胞;生殖毒性;Bax;Bcl-2

Abstract:

Keywords:

基金项目(Foundation): 内蒙古自治区2012年自然科学基金(2012MS1146);; 2015年内蒙古医科大学青年创新基金(dncx201506)

作者(Authors): 董瑞娜;刘殊;张敏;戴红;徐肖倩;

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