title: [Expression of alpha 1 anti-trypsin in proximal tubular epithelial cell line].
abstract: OBJECTIVE
  To investigate the expression and regulation of alpha 1 anti-trypsin (AAT) in tubular epithelial cells.
  
  
  METHODS
  The expression of AAT in tubular epithelial cell line HKC was detected with indirect immunofluorescence assay and confirmed by reverse transcription polymerase chain reaction (PCR) in transcription level respectively, and PCR product was sequenced. In order to observe the response of HCK to LPS, the different concentrations of LPS (0, 0.5, 1.0, 2.0 microg/ml) were used in the research and the regulation of AAT in HKC was semi-quantitatively detected with Western Blot and Real-Time PCR respectively.
  
  
  RESULTS
  Indirect immunofluorescence staining showed that AAT was positive in HKC, but negative in renal interstitial fibroblast. By PCR, a significant messenger RNA band of AAT was found in HKC, but not in renal interstitial fibroblast. DNA sequencing indicated that the sequence of PCR product is consistent with AAT mRNA sequence in Gene Bank. Real-time PCR showed that the expression of AAT mRNA was up-regulated (fluorescence intensity ratio is 3.43 +/- 0.88 versus 1.22 +/- 0.20; P < 0.01) in HKC stimulated with 2.0 microg/ml LPS for 4 h, and Western Blot showed that the synthesis of AAT significantly increased (band density ratio is 0.88 +/- 0.12 versus 0.59 +/- 0.05; P < 0.01) in HKC stimulated with 2.0 microg/ml LPS for 8 h, as compared with unstimulated HKC. 0.5, 1.0 g/ml of LPS has no effect on the expression of AAT mRNA and AAT protein in HKC.
  
  
  CONCLUSIONS
  HKC express AAT and the expression can be up-regulated by LPS.
authors.affiliation: The Center of Nephrology, China-Japan Friendship Hospital, Beijing 100029, China.
authors.forename: Gong-yao
authors.initials: GY
authors.lastname: Tang
authors.forename: Yi-fu
authors.initials: YF
authors.lastname: Chen
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.forename: Hong-liang
authors.initials: HL
authors.lastname: Rui
authors.forename: Xiao
authors.initials: X
authors.lastname: Cong
authors.forename: Jin-ning
authors.initials: JN
authors.lastname: Lou
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis*
chemical_list: D008070:Lipopolysaccharides; D012333:RNA, Messenger; D000515:alpha 1-Antitrypsin