Overexpression of heat shock protein Hsp27 protects against cerulein-induced pancreatitis.
OBJECTIVE
Heat shock protein (Hsp) 27 regulates actin cytoskeletal dynamics, and overexpression of Hsp27 in fibroblasts protects against stress in a phosphorylation-dependent manner. Induction of Hsps occurs in acute pancreatitis, but Hsp27 has not been ascribed a specific role. To examine whether Hsp27 would ameliorate acute pancreatitis, we generated transgenic mice overexpressing human Hsp27 (huHsp27) or Hsp27 with the phosphorylatable residues Ser(15,78,82) mutated to aspartic acid (huHsp27-3D) to mimic phosphorylation or to alanine (huHsp27-3A), which is nonphosphorylatable.


METHODS
huHsp27 was expressed at high levels in the exocrine pancreas by use of a cytomegalovirus promoter. Protein expression was analyzed by Western blotting and immunofluorescence. Acute pancreatitis was induced with 6 or 12 hourly cerulein injections (50 microg/kg intraperitoneally) and its severity assessed by measuring serum amylase and lipase levels, pancreatic trypsin activity, edema, and morphologic changes by quantitative scoring of multiple histologic sections and visualization of filamentous actin. Systemic inflammatory effects were monitored by measuring lung myeloperoxidase activity (a marker of neutrophil infiltration).


RESULTS
huHsp27 protein was overexpressed in the pancreas and localized to pancreatic acini. Acute pancreatitis was ameliorated by overexpression of huHsp27 and the huHsp27-3D mutant, which were associated with suppression of pancreatic trypsin activity and acinar cell injury and preservation of the actin cytoskeleton. In contrast, these changes were unaffected by overexpression of the nonphosphorylatable huHsp27-3A mutant.


CONCLUSIONS
Pancreatic overexpression of huHsp27 protects against cerulein-induced acute pancreatitis in a specific phosphorylation-dependent manner and is associated with preservation of the actin cytoskeleton.
Kubisch
Constanze
C
Department of Internal Medicine II, Klinikum Grosshadern, Ludwig-Maximilians University Munich, Germany.
Dimagno
Matthew J
MJ
Tietz
Anne Barbara
AB
Welsh
Michael J
MJ
Ernst
Stephen A
SA
Brandt-Nedelev
Barbara
B
Diebold
Joachim
J
Wagner
Andreas C C
AC
Göke
Burkhard
B
Williams
John A
JA
Schäfer
Claus
C
D000199:Actins / Q000187:drug effects / Q000276:immunology; D000208:Acute Disease; D000818:Animals; D002108:Ceruletide / Q000009:adverse effects*; D003599:Cytoskeleton / Q000187:drug effects / Q000276:immunology; D005260:Female; D005765:Gastrointestinal Agents / Q000009:adverse effects*; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D023421:Models, Animal; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D010195:Pancreatitis / Q000139:chemically induced / Q000276:immunology*; D010766:Phosphorylation / Q000187:drug effects; D012720:Severity of Illness Index
D000199:Actins; D005765:Gastrointestinal Agents; D006360:Heat-Shock Proteins; C486113:Hsbp1 protein, mouse; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D002108:Ceruletide