Expression And Significance Of Calpain2in Obstructive Jaundice Rats And The Intervention Role Of Traditional Chinese Medicine

Objective:In recent years, the study found that endoplasmic reticulum stress is an important way to cause apoptosis. Whether calpain2in the endoplasmic reticulum stress mediated liver cell apoptosis play an important role in the process, and whether been used Traditional Chinese Medicine by endoplasmic reticulum stress signaling pathways regulate the apoptosis, has not been reported.Methods:Choose adult SD rats weight (250-270),160g, male, animal level to clean, random group of rats:(1) sham operation group (S)40.(2) obstructive jaundice group (O group)40.(3) obstructive jaundice model+medicine administered group (OC group)40.(4) model of obstructive jaundice+saline group (OW group)40.Four groups respectively at1,3,5,7d, each phase to be put to death in the rat10, only to return in serum and liver tissue. In the detection of serum TBIL. ALT. AST and biochemical indexes such as content; Another liver tissues were fixed in5%neutral formalin solution, embedded in paraffin.5μm sections were HE staining of pathological changes in liver tissue. And immunohistochemical detection of liver tissue calpain2, expression. After remaining liver tissue frozen in liquid nitrogen at-80℃, measured by western blot liver tissue calpain2expression. Detection of m-calpain gene expression of rat liver tissue case with Real-Time PCR.Results:1. The serum biochemical indicators of change:after the first day, ALT, AST increased slightly, then gradually decreased to the normal range, the difference was not statistically significant in S group. Compared with the group S, ALT, AST increased rapidly after treatment in the O group, and ALT increased rapidly, and maintain a high level, after3days were decreased (P<0.05). ALT, AST were elevated in OC group and the OW group. And all peaked on the first day after surgery, after gradually reduce. At various time points,ALT, AST values in OC group were lower than in OW group. TBIL in S group were no significant changes after surgery, and there was no statistically significant difference. Compared with the group S, TBIL in O group increased rapidly after bile duct ligation. and reached a peak (P <0.05) in the first three days, after a slow decline. TBIL groups were increased significantly, the rapid increase in the value of postoperative TBIL. and peaked in the third day, after a slow decline.OC group were lower than OW group at each time point TBIL value.2. Liver tissue pathology change: No swelling of liver cells, necrosis, hepatic cell cord neatly arranged in S group. Group O of visible liver tissue inflammatory cells infiltration, small bile ducts and the lumen of small bile duct hyperplasia, fiber tissue hyperplasia, degeneration necrosis of hepatic cells. OC group than in terms of OW group, inflammatory cell infiltration in the liver tissue less fibrous tissue proliferation lighter, less liver cell degeneration and necrosis.3. Using western blot test calpain2protein expression changes in liver tissue:At the protein level, for GAPDH internal, S group as control group, O set of calpain2protein content and comparison of each phase had a significant difference (P<0.05), and O group calpain2protein levels higher than the S group. OW group as control group, the OC group calpain2protein content and comparison of various phase had a significant difference (P<0.05), and OC group calpain2protein expression levels below OW group. O, OW, OC group as the building time prolonged, calpain2protein expression are on the rise.4. Immunohistochemical detection of liver tissue calpain2expression:Then, the normal group rats less calpain2protein expression in the liver. As building the extension of time, the O group, OW, OC group of calpain2protein positive expression quantity increase gradually, and is mainly expressed in cytoplasm of liver cells, occasional expressed in the nuclei. Compared with normal control group (P<0.05).5、Detection of m-calpain gene expression of rat liver tissue case with Real-Time PCR.:Group S m-calpain gene expression level with extended building time has no obvious change. Compared with the group S O group, O group m-calpain gene expression levels in various time points were higher than S group (P<0.05). OW group, OC group m-calpain gene expression levels increased with longer time modeling. Compared with the group S OW group, OC group m-calpain gene expression levels in various time points were lower than OW group (P<0.05).Conclusions:1. When obstructive jaundice, with the extension of modeling time, calpain2protein increased gradually, indicating that with the extension of the modeling of time, gradually increase the apoptotic index.2. Obstructive jaundice rats for Traditional Chinese Medicine can inhibit the artemisia capillaris soup calpain2protein expression, reduce caused by high calpain2protein expression of liver cell injury, improve liver caused by obstruction of abnormal liver function, and m-calpain gene expression levels consistent with obstructive jaundice liver damage degree change, suggesting m-calpain participates in obstructive jaundice liver endoplasmic reticulum stress apoptosis-mediated cell occurrence and development process. Dialectical, clinical treatment of patients with obstructive jaundice can use the formula, promote the recovery of liver function, reduce the incidence of complications.