title: [Protein kinase C-nuclear factor-erythroid 2-related factor 2 regulating the expression of heme oxygenase-1 in rat airway epithelial cells].
abstract: OBJECTIVE
  To observe the effect of the signaling pathway of protein kinase C (PKC)-nuclear factor-erythroid 2-related factor 2 (Nrf2) on the expression of heme oxygenase -1 (HO-1) induced by cigarette smoke extract in rat airway epithelial cells.
  
  
  METHODS
  The airway epithelial cells of 25 male SD rats were randomly divided into a control group, a CSE3h group, a RO318220 group (PKC inhibitor), a Nrf2 siRNA group and a Nrf2 siRNA+RO318220 group, 5 rats in each group. The control group was incubated with DMEM/F12 alone. The CSE3h group was treated with 10% CSE for 3 h. The RO318220 group was pretreated with 3 μmol/L RO318220 for 0.5 h and subsequently treated with 10% CSE for 3 h. The Nrf2 siRNA group was pretreated with Nrf2 siRNA, and then treated with 10% CSE for 3 h. The Nrf2 siRNA+RO318220 group was pretreated with Nrf2 siRNA and 3 μmol/L RO318220 for 0.5 h, and then treated with 10% CSE for 3 h. The protein levels of Nrf2 in the nucleus and cytoplasm, and HO-1 and PKC in the whole cells were semi-quantified by Western blot. The protein expression of HO-1 was measured by immunocytochemistry. HO-1 mRNA expression was detected by RTPCR. Immunofluorescence staining was used to observe the nuclear translocatin of Nrf2.
  
  
  RESULTS
  CSE markedly induced Nrf2 nuclear translocation in the rat airway epithelial cells, and RO318220 pretreatment blocked the CSE induced Nrf2 nuclear translocation. Immunocytochemistry showed that HO-1 protein expression was strongly positive in the CSE3h group, weakly positive in the other 4 groups. The expression of PKC protein, HO-1 mRNA and protein significantly increased in the CSE3h group, and HO-1 activity markedly improved in the CSE group (P<0.05). The level of PKC protein expression was not significantly different in the Nrf2 siRNA group compared with that in the CSE3h group (P>0.05).
  
  
  CONCLUSIONS
  CSE induces the nuclear translocation of Nrf2 by PKC signaling pathway, thus upregulating the HO-1 expression in the rat airway epithelial cells.
authors: Department of Respiratory Medicine, Hunan Provincial People's Hospital, Changsha 410005, China jianggang68@163.com.
authors: Gang
authors: G
authors: Jiang
authors: Zhiguang
authors: Z
authors: Liu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism*; D007211:Indoles; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D021381:Protein Transport; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D012906:Smoke; D015854:Up-Regulation
chemical_list: D007211:Indoles; D051267:NF-E2-Related Factor 2; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012906:Smoke; D006419:Heme Oxygenase (Decyclizing); C491523:Hmox1 protein, rat; D011493:Protein Kinase C; C064758:Ro 31-8220