过氧化物酶体增殖物激活受体α基因在全氟辛酸致大鼠肝BRL-3A细胞氧化损伤中的作用  

Role of PPARα in the oxidative damage of rat liver cells BRL-3A induced by perfluorooctanoic acid

作　　者：王力[1] 胡子琰 王伟业 闻赵燕 阚梦颖 刘辉[1] Wang Li;Hu Ziyan;Wang Weiye;Wen Zhaoyan;Kan Mengying;Liu Hui(Bengbu Medical College,Bengbu 233030,Chin)

机构地区：[1]蚌埠医学院,蚌埠233030

出　　处：《卫生研究》2018年第3期465-470,共6页Journal of Hygiene Research

基　　金：国家自然科学基金(No.81703227);安徽省自然科学基金(No.1508085QH188);蚌埠医学院科技发展基金(No.BYKF1601);安徽省大学生创新训练计划(No.201610367010;201610367058)

摘　　要：目的通过抑制和激活基因表达的方式,研究过氧化物酶体增殖物激活受体α基因(PPARα)在全氟辛酸(PFOA)致大鼠肝BRL-3A细胞氧化损伤中的作用。方法体外培养大鼠肝BRL-3A细胞,分为空白对照组(NC)、PFOA实验对照组、PPARα抑制剂组(GW6471)、PPARα激动剂组(WY14643)、PPARα抑制剂预处理PFOA组(GW6471+PFOA)、PPARα激动剂预处理PFOA组(WY14643+PFOA)。荧光免疫细胞化学检测法检测基因抑制和激动表达情况;自由基指示剂H2DCFDA检测活性氧(ROS)含量;q PCR检测PPARα及其下游基因Cyp4a1的表达;Western blot检测PPARα蛋白表达水平。结果通过抑制剂和激动剂成功抑制和激动了PPARα在大鼠肝BRL-3A细胞中的表达。GW6471+PFOA组与空白对照组、PFOA实验对照组比较,细胞内ROS含量显著升高(P<0.05);WY14643+PFOA组与空白对照组、PFOA实验对照组相比,ROS含量明显下降(P<0.05)。PPARα及其下游基因Cyp4a1在GW6471+PFOA组中的表达比PPARα抑制剂组高,但相较PFOA实验对照组明显降低(P<0.05)。WY14643+PFOA组相关基因的表达虽然低于PPARα激动剂组,但显著高于PFOA实验对照组(P<0.05)。GW6471+PFOA组中PPARα蛋白的表达水平较抑制剂组有所上调,但与空白对照组相比差异无统计学意义。WY14643+PFOA组中PPARα蛋白的表达水平与激动剂组比差异无统计学意义,但却显著高于PFOA实验对照组(P<0.05)。结论 PFOA的暴露可以激活PPARα的表达,减轻细胞内ROS的累积,PPARα在PFOA导致的大鼠肝脏细胞氧化损伤中起到了一定的保护作用。Objective To investigate the role of PPARα in oxidative damage of BRL-3A cells induced by perfluorooctanoic acid（ PFOA） by inhibiting and activating gene expression. Methods In vitro culture of rat liver BRL-3A cells were divided into blank control group,PFOA experimental control group,PPARα inhibition group（ GW6471）,PPARα agonist group（ WY14643）, PPARα inhibitor pretreatment PFOA group（ GW6471 ＋ PFOA）,PPARα agonist pretreatment PFOA group（ WY14643 ＋ PFOA）.Fluorescence immunocytochemistry was used to detect the expression of PPARα. The expression of PPARα and its downstream target gene was detected by q PCR. The expression of related protein was detected by Western blot. Results The expression of PPARα in rat liver BRL-3A cells was successfully inhibited and stimulated by inhibitors and agonists（ P〈0. 05）. Compared with the blank control group and the PFOA experimental control group,there was a significant decrease in the content of ROS in the WY14643 ＋ PFOA group compared with the blank control group and the PFOA experimental control group（ P〈0. 05）. The expression of PPARα and its downstream gene Cyp4a1 in GW6471 ＋ PFOA group was higher than that in PPARα inhibitor group（ P〈0. 05）,but it was significantly lower than that in PFOA experimental control group（ P〈0. 05）. The expression of related genes in WY14643 ＋ PFOA group was significantly lower than that in PPARα agonist group（ P〈0. 05）. The protein expression of PPARα in GW6471 ＋ PFOA group was up-regulated compared with the inhibitor group,there was no difference compared with the blank control group. The protein expression of PPARα in WY14643 ＋ PFOA group was not significantly different from that in agonist group,but it was significantly higher than that in PFOA experimental control group（ P〈0. 05）.Conclusion PFOA exposure can activate the expression of PPARα,remove ROS,PPARα played a protective role in PFOA-induced rat liver cell oxidative damage.

关 键 词：全氟辛酸 氧化损伤 过氧化物酶体增殖物激活受体Α 肝脏细胞 

分 类 号：Q55[生物学—生物化学] Q593.1