title: Altered expression levels of occludin, claudin-1 and myosin light chain kinase in the common bile duct of pediatric patients with pancreaticobiliary maljunction.
abstract: BACKGROUND
  In pancreaticobiliary maljunction (PBM), the sphincter of Oddi can not control bile and pancreatic juice flow, which may lead to two-way reflux of bile and pancreatic juice, thus causing chronic inflammation, thickening, fibrosis and metaplasia of the common bile duct wall. These pathophysiological changes have been linked to disruption of the epithelium barrier in the common bile duct. We hypothesized that the expression of tight junction-associated proteins may be dysregulated in the common bile duct in PBM. In the current study, we sought to analyze the expression of tight junction-associated proteins in the common bile duct epithelium of pediatric patients with PBM.
  
  
  METHODS
  Specimens of the common bile duct were collected from 12 pediatric patients with PBM and 10 non-PBM controls. The expression of the tight junction-associated proteins occludin and claudin-1 in the epithelium was examined by immunohistochemistry. The Image-Pro Plus v. 6.0 image analysis software was used to calculate the mean qualifying score (MQS) of imunostained sections of common bile duct epithelium. Total protein extracts of common bile duct were analyzed by Western blotting assays to examine expression of occludin, claudin-1 and myosin light chain kinase (MLCK). Spearman correlation analysis was used to analyze the relation between MLCK and occludin, MLCK and claudin-1.
  
  
  RESULTS
  Immunostained sections of the common bile duct epithelium showed significantly higher MQS in pediatric patients than controls for occludin (44.11 ± 13.82 vs. 11.30 ± 9.58, P = 0.0034) and claudin-1 (63.44 ± 23.59 vs. 46.10 ± 7.84, P = 0.0384). Western blotting also showed significantly higher expression of occludin, claudin-1 and MLCK in the common bile duct of patients than of controls (P = 0.0023, 0.0015, 0.0488). Spearman correlation analysis showed that MLCK expression correlated positively with the expression of occludin (r s = 0.61538, P = 0.0032) and claudin-1 (r s = 0.7972, P = 0.0019).
  
  
  CONCLUSIONS
  Occludin and claudin-1 are up-regulated in the common bile duct epithelium of pediatric PBM patients. MLCK may be involved in the process of up-regulation of the tight junction-associated proteins in PBM.
authors.affiliation: Departments of Pediatric General Surgery, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China.
authors.forename: Shun-Gen
authors.initials: SG
authors.lastname: Huang
authors.affiliation: Department of Radiology, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China. gwlsuzhou@163.com.
authors.forename: Wan-Liang
authors.initials: WL
authors.lastname: Guo
authors.affiliation: Departments of Pediatric General Surgery, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China.
authors.forename: Zhi-Cheng
authors.initials: ZC
authors.lastname: Zhou
authors.affiliation: Departments of Pediatric General Surgery, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China.
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Li
authors.affiliation: Department of Radiology, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China.
authors.forename: Fu-Bin
authors.initials: FB
authors.lastname: Yang
authors.affiliation: Departments of Pediatric General Surgery, Children's Hospital affiliated to Soochow University, Suzhou, Jiangsu, 215003, China. wangjiansuzhou@163.com.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
mesh_terms: D001652:Bile Ducts / Q000002:abnormalities*; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D062445:Claudin-1 / Q000378:metabolism*; D003135:Common Bile Duct / Q000378:metabolism*; D004848:Epithelium / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D008297:Male; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D062793:Occludin / Q000378:metabolism*; D010183:Pancreatic Ducts / Q000002:abnormalities*; D015854:Up-Regulation
chemical_list: C568108:CLDN1 protein, human; D062445:Claudin-1; C568985:OCLN protein, human; D062793:Occludin; D009219:Myosin-Light-Chain Kinase