Effect of Tetrandrine on LPS-induced NF-kappaB activation in isolated pancreatic acinar cells of rat.
OBJECTIVE
To investigate the effect of Tetrandrine (Tet) on LPS-induced NF-kappaB activation and cell injury in pancreatic acinar cells and to explore the mechanism of Tetrandrine preventing LPS-induced acinar cell injury.


METHODS
Male rat pancreatic acinar cells were isolated by collagenase digestion, then exposed to LPS (10 mg/L), Tet (50 micromol/L, 100 micromol/L) or normal media. At different time point (30 min, 1 h, 4 h, 10 h) after treatment with the agents, cell viability was determined by MTT, the product and nuclear translocation of subunit p65 of NF-kappaB was visualized by immunofluorescence staining and nuclear protein was extracted to perform EMSA which was used to assay the NF-kappaB binding activity.


RESULTS
LPS induced cell damage directly in a time dependent manner and Tet attenuated LPS-induced cell damage (50 micromol/L, P < 0.05; 100 micromol/L, P < 0.01). NF-kappaB p65 immunofluorescence staining in cytoplasm increased and began showing its nuclear translocation within 30 min and the peak was shown at 1 h of LPS 10 mg/L treatment. NF-kappaB DNA binding activity showed the same alteration pattern as p65 immunofluorescence staining. In Tet group, the immunofluorescence staining in cytoplasm and nuclear translocation of NF-kappaB were inhibited significantly.


CONCLUSIONS
NF-kappaB activation is an important early event that may contribute to inflammatory responses and cell injury in pancreatic acinar cells. Tet possesses the protective effect on LPS-induced acinar cell injury by inhibiting NF-kappaB activation.
Zhang
Hong
H
Department of Pathophysiology, Medical College of Tongji University, Shanghai 200092, China. zhangh1227@sohu.com
Li
Yong-Yu
YY
Wu
Xian-Zhong
XZ
D000208:Acute Disease; D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D044182:Benzylisoquinolines / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D008070:Lipopolysaccharides / Q000009:adverse effects / Q000494:pharmacology*; D008297:Male; D016328:NF-kappa B / Q000187:drug effects* / Q000378:metabolism; D010179:Pancreas / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010195:Pancreatitis / Q000188:drug therapy; D051381:Rats; D017207:Rats, Sprague-Dawley
D000470:Alkaloids; D044182:Benzylisoquinolines; D002121:Calcium Channel Blockers; D008070:Lipopolysaccharides; D016328:NF-kappa B; C009438:tetrandrine