title: [The pathway of PI3k/Akt-aPKCiota/zeta-Nrf2 regulating the expression of gamma-glutamylcysteine synthetase in the bronchial epithelial cells of rats].
abstract: OBJECTIVE
  To observe the effect of the signal pathway of phosphoinositol-3-kinase (PI3K)/Akt-antypical protein kinase C(iotazeta) (aPKC(iotazeta))-Nuclear factor-E2 related factor (Nrf2) on gamma-glutamylcysteine synthetase (gamma-GCS) of the bronchial epithelial cells of rats after exposure to cigarette smoke extracts (CSE).
  
  
  METHODS
  gamma-GCS, Nrf2, p-Akt and p-aPKC(iotazeta) proteins were semi-quantified by Western blot. gamma-GCS protein expression was assessed by immunocytochemistry. gamma-GCS mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Nrf2 protein was observed by immunofluorescence. The rate of the cells expressed p-Akt were analyzed by flow cytometry. GSH content and gamma-GCS activity were measured.
  
  
  RESULTS
  GSH content, Nrf2 protein of nucleus, p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity were significantly increased after exposure to CSE for 3 hours. aPK(iotazeta) inhibitor RO813220 significantly reduced the expression of p-aPKC(iotazeta) protein, gamma-GCS protein and mRNA and activity, but enhanced Nrf2 protein of cytoplasm expression, had no effect on p-Akt. p-Akt inhibitor LY294002 and RO813220 + LY294002 decreased p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity expression, increased Nrf2 protein of cytoplasm expression. The correlation analysises demonstrated that there were a positive correlation between Nrf2 and gamma-GCS, p-Akt, p-aPKC(iotazeta), between p-Akt and Nrf2, p-aPKC(iotazeta), gamma-GCS, between p-aPKC(iotazeta) and Nrf2, p-Akt, gamma-GCS.
  
  
  CONCLUSIONS
  CSE might upregulate gamma-GCS expression through PI3K/Akt-aPKC(iotazeta)-Nrf2 signaling pathway in the bronchial epithelial cells of rats.
authors.affiliation: Department of Respiratory, Hunan Provincial People's Hospital, Changsha 410005, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Jiang
authors.forename: Ai-Guo
authors.initials: AG
authors.lastname: Dai
mesh_terms: D000818:Animals; D008297:Male; D051381:Rats; D001980:Bronchi / Q000166:cytology / Q000201:enzymology; D004781:Environmental Exposure / Q000009:adverse effects; D004847:Epithelial Cells / Q000201:enzymology; D005721:Glutamate-Cysteine Ligase / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D051058:Oncogene Protein v-akt / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014028:Tobacco Smoke Pollution / Q000009:adverse effects; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D000099293:Protein Kinase C-lambda
chemical_list: D005721:Glutamate-Cysteine Ligase; D007527:Isoenzymes; D051058:Oncogene Protein v-akt; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C; D012333:RNA, Messenger; D014028:Tobacco Smoke Pollution; D051267:NF-E2-Related Factor 2; D000099293:Protein Kinase C-lambda