摘要
目的:探讨蛋白激酶C(PKC)-红系衍生的核因子相关因子2(Nrf2)对香烟烟雾提取物(CSE)诱导的大鼠气道上皮细胞血红素加氧酶1(HO-1)表达的影响。方法:通过CSE刺激雄性SD大鼠气道上皮细胞,使用PKC抑制剂RO318220和Nrf2 siRNA,将细胞分为对照组、CSE 3 h组、RO318220组、Nrf2 siRNA组和RO318220+Nrf2 siRNA组,用Western blotting法分别检测HO-1、Nrf2和p-PKC蛋白表达,免疫细胞化学法观察HO-1蛋白表达,逆转录-聚合酶链反应(RT-PCR)法检测HO-1 mRNA表达,免疫荧光法检测Nrf2蛋白定位,测定HO-1活性。结果:暴露CSE 3 h后,Nrf2蛋白主要表达在胞核,胞核蛋白表达增强,p-PKC蛋白、HO-1的mRNA和蛋白高表达,HO-1活性增强。预先给予RO318220,PKC蛋白、Nrf2胞浆和胞核蛋白、HO-1的mRNA和蛋白表达均明显减弱,HO-1活性显著降低。预先用siRNA沉默Nrf2,胞浆和胞核的Nrf2蛋白表达均减弱,HO-1活性、mRNA和蛋白水平明显降低。RO318220联合Nrf2 siRNA处理后,PKC蛋白、Nrf2胞浆和胞核蛋白、HO-1 mRNA和蛋白表达均明显降低,HO-1活性明显降低。结论:CSE通过PKC激活Nrf2,诱导Nrf2核转位,从而上调HO-1的表达水平。
[ABSTRACT]AIM：ToobservetheeffectofthesignalingpathwayofproteinkinaseC（PKC）-nuclearfactor-ery-throid 2-related factor 2 （Nrf2） on the expression of heme oxygenase-1 （HO-1） induced by cigarette smoke extract （CSE） in the rat airway epithelial cells .METHODS：After exposed to CSE , the airway epithelial cells of male SD rats were di-vided into control group , CSE3 group, RO318220 group, Nrf2 siRNA group and RO318220＋Nrf2 siRNA group.The pro-tein levels of HO-1, Nrf2 and PKC were semi-quantified by Western blotting .The protein expression of HO-1 was assessed by immunocytochemistry .The mRNA expression of HO-1 was detected by reverse transcription-polymerase chain reaction （ RT-PCR） .Nrf2 protein location was observed by immunofluorescence staining .HO-1 activity was also determined .RE-SULTS：After exposure to CSE for 3 h, the Nrf2 protein was mainly located in the nucleus , and the expression level of Nrf2 protein in the nucleus was stronger than that in the control cells .CSE also significantly enhanced the levels of p-PKC protein, and HO-1 mRNA, protein and activity .Pretreatment with RO318220 significantly decreased the levels of PKC pro-tein, Nrf2 protein, and HO-1 mRNA, protein and activity.Meanwhile, Nrf2 protein significantly decreased , and the activ-ity, mRNA and protein levels of HO-1 were also significantly attenuated by pretreatment with siRNA to knock down Nrf2. Pretreatment with RO318220 and Nrf2 siRNA significantly decreased the PKC protein , Nrf2 protein, and HO-1 mRNA, protein and activity .CONCLUSION：CSE up-regulates the HO-1 expression through PKC-induced nuclear translocation of Nrf2 in the rat airway epithelial cells .
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第8期1483-1488,共6页
Chinese Journal of Pathophysiology
关键词
香烟烟雾提取物
红系衍生的核因子相关因子2
血红素加氧酶1
蛋白激酶C
Cigarette smoke extract
Protein kinase C Nuclearfactor-erythroid 2-relatedfactor 2 Heme oxy-genase-1
