Expression Of IRE1 In Vitro Liver Cell Models With Serum Of Rats With Obstructive Jaundic And The Mechanism Of Traditional Chinese Medicine

Objective:To investigate the difference of IRE1 expression in liver cells of rats after a ddingdifferent serum. Discussing the mechanism of liver injury in obstructive jaun dice and the protection mechanism of Yinchenhao Decoction. To provide a theore tical basis for reasonable application of traditional Chinese medicine. Methods:1.SD rat 30,adult and male,270-300 g,animal level of clean grade,were rando mly divided into 3 groups:1.the obstructive jaundice group(Group O,N=10); 2.the co ntrol group(Group S,N=10);3. The feeding of Yinchenhao Decoction group(Gro up Y,N=10);.Group S:line control operation, separation bile duct but not ligatio n;O group:separation and double ligation bile duct; Y group: intragastric admin istration with Yinchenhao Decoction(dose 1m L/100g) by 1 times a day. All th e rats in this three groups were sacrificed on the 5 day after the operation.The blood were harvested from abdominal aorta.Part of the blood was used to the detection of serum ALT,AST,TBIL and DBIL; The others were retention of se rum after centrifugation.2. Rat normal liver cell line BRL-3A was selected, and purchased from the cell bank of Chinese Academy of Sciences. After cultured in vitro, the species were divided into three groups, A, B and C.(1)A group was added to the no rmal rat serum containing mediumthe;(2)B group to join the obstructive jaundi ce rat serum containing medium;(3)C group for with obstruction jaundice rats s erum and feeding Yinchenhao Decoction in rat serum.Getting the others part of the liver cells and culture supernatant in 6h, 24 h, 48h; Labeled as A6 h, A24 h, A48h, B6 h, B24 h, B48 h, C6 h, C24 h, C48 h.hemistry, Western blot was used t o determine the changes of IRE1? protein expression in liver cells. RT-PCR w as used to determine the changes of IRE1? m RNA.The culture solution used t o be testing content of ALT and AST. Results:1.Change of serum biochemical indicators:(1)Blood serum of experimental animal model: AST,ALT, TBIL and DBIL in groups O significantly increased c ompared with control group S(P<0.05), The establishment of a rat model of o bstructive jaundice was successful. And we have got the necessary blood seru m.(2)Cell culture assay: the A group as the control group, B group and C gro up ALT, AST level at each time points were higher than the A group(P<0.05); compared with C, B group of ALT, AST level at all time points were low er than the B group(P<0.05).2.Western blot detection of liver cells IRE1? expression:taking Actin as the r eference, the IRE1? expression level of B group and C group at each time poi nts were higher than the A group(P<0.05); compared with B group, the IRE1? expression level of C group at all time points were lower than the B group(P<0.05).3.RT-PCR method to detect IRE1? m-RNA expression: The IRE1? m-RNA expression level of B group and C group at each time points were higher tha n the A group(P<0.05); compared with B group, the IRE1? m-RNA expressio n level of C group at all time points were lower than the B group(P<0.05). Conclusions:1. The normal rat liver cells adding the serum of rats with obstructive jaund ice can be successfully manufactured in the liver injury model. ERS apoptosis of IRE1?-JNK pathway is a possible mechanism of liver injury induced by ob structive jaundice.2. Adding Yinchenhao Decoction containing serum can improve liver functio n in the liver injury model of obstructive jaundice. It may be through inhibitin g the excessive activation of IRE1? protein and reducing the ERS reaction.3. According to the perioperative management of OJ,a rational operation way combining with clearing away dampness-heat and removing dampness herbs ca n promote early recovery of liver function, reduce the complication incidence of OJ, and lead to better therapeutic effect for clinical.