Cigarette smoke extract regulates expression of heme oxygenase-1 through signaling pathway of protein kinase C-nuclear factor-erythroid 2-related factor 2 in rat airway epithelial cells
10.3969/j.issn.1000-4718.2014.08.025
- VernacularTitle:香烟烟雾提取物通过蛋白激酶 C-核因子相关因子2调节大鼠气道上皮细胞血红素加氧酶1表达
- Author:
Gang JIANG
;
Weidong ZHANG
- Publication Type:Journal Article
- Keywords:
Cigarette smoke extract;
Protein kinase C;
Nuclearfactor-erythroid 2-relatedfactor 2;
Heme oxy-genase-1
- From:
Chinese Journal of Pathophysiology
2014;(8):1483-1488
- CountryChina
- Language:Chinese
-
Abstract:
[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 .