[过氧化物酶体增殖物激活受体α在全氟辛酸诱导的大鼠肝细胞BRL-3A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].
作者信息
Wang Li, Hu Ziyan, Wang Weiye, Wen Zhaoyan, Kan Mengying, Liu Hui
机构信息
Bengbu Medical College, Bengbu 233030, China.
出版信息
Wei Sheng Yan Jiu. 2018 May;47(3):465-470.
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.
目的
通过抑制和激活基因表达，探讨过氧化物酶体增殖物激活受体α（PPARα）在全氟辛酸（PFOA）诱导的BRL-3A细胞氧化损伤中的作用。
方法
体外培养大鼠肝脏BRL-3A细胞，分为空白对照组、PFOA实验对照组、PPARα抑制组（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）。采用荧光免疫细胞化学法检测PPARα的表达。采用qPCR检测PPARα及其下游靶基因的表达。采用蛋白质免疫印迹法检测相关蛋白的表达。
结果
抑制剂和激动剂成功抑制和刺激了大鼠肝脏BRL-3A细胞中PPARα的表达（P<0.05）。与空白对照组和PFOA实验对照组相比，WY14643+PFOA组活性氧（ROS）含量显著降低（P<0.05）。GW6471+PFOA组PPARα及其下游基因Cyp4a1的表达高于PPARα抑制剂组（P<0.05），但显著低于PFOA实验对照组（P<0.05）。WY14643+PFOA组相关基因的表达显著低于PPARα激动剂组（P<0.05）。GW6471+PFOA组PPARα蛋白表达较抑制剂组上调，与空白对照组相比无差异。WY14643+PFOA组PPARα蛋白表达与激动剂组无显著差异，但显著高于PFOA实验对照组（P<0.05）。
结论
PFOA暴露可激活PPARα的表达，清除ROS，PPARα在PFOA诱导的大鼠肝细胞氧化损伤中起保护作用。
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