Study Of The Expression And Relationship Of MLCK,Cacium Channel Protein In Hypertriglyceridemia Associated Acute Pancreatitis

Objective To observe the changes of pathological scores of pancreatitis,serum amylase(AMY),ultrastructure and tight junction(TJ),and detect the expression of MLCK,Cav1.2,Cav2.1and Cav2.2 protein in pancreas in HTGP and to investigate the expression and relationship of MLCK and cacium channel in rats with hypertriglyceridemia-associated acute pancreatitis.Methods Forty-eight male Sprague Dawley rats were randomly divided into normal diet group and high fat diet group.After being raised for 4weeks,blood was measured serum TG level.Then normal diet group were randomly divided into three subgroups: N group,AP group and AP+ML-7group;high fat diet group were randomly divided into three subgroups: HTG group,HTGP group and HTGP+ML-7 group.AP was induced by intraperitoneal injection of cerulean(50μg/kg).The rats were sacrificed at 24 hours after AP induction.Serum amylase(AMY)was measured.At the same time,the pathological changes of pancreas in rats were observed under light microscope.The ultrastructure and tight junction(TJ)change in pancreas were observed withtransmission electron microscopy.The expression and localization of MLCK and calcium channel in the pancreas were assayed by immunohistochemistry and immunofluorescence.Results The levels of TG in normal diet group and high fat diet group were respectively(1.16+0.33)mmol/L and(3.30+1.07)mmol/L,the levels of TG in high fat diet group was significantly higher than in normal diet group(P<0.01).After AP was performed,that AMY level and pathological score in pancreas were increased.AMY level and pathological score in HTGP group were significantly higher than in AP group(P<0.05).ML-7 can decrease serum amylase level and pathological score.The expression of MLCK and Cav1.2,Cav2.2 and Cav2.2 in HTGP group were significantly higher than those in AP group(P<0.05).ML-7 can effectively decrease the expression of MLCK and Cav1.2,Cav2.1 and Cav2.2.Compared with AP group,the ultrastructure of pancreas group was more significantly damaged and TJ was more seriously broadened in HTGP group with transmission electron microscopy.ML-7 can significantly improve TJ.Conclusion The expression of MLCK and Cav1.2,Cav2.1,Cav2.2in pancreas of rats was up-regulated,which may be related to the severity of HTGP.MLCK may be involved in the pathogenesis of HTGP by broadening TJ and regulating the expression of cacium channel.