The Effection Of Jie Du Tongluo Decoction On Endoplasmic Reticulum Stress Through The IRE1?/JNK Pathway

Purpose:To observe the effect of JieduTonluotiaogan Decoction on improving IR in T2DM model rats and the expression of inflammatory factors and protein in HepG2 cells stimulated by tunicamycin.Methods:1.The animal model of type 2 diabetic rats was reproduced by feeding ZDF rats with high fat pellets Purina#5008.JieduTonluotiaogan Decoction?high,medium and low concentration?was applicated to cure these rats,and metformin was used as a positive drug to observe the expression of IRE1,P-IRE1,JNK and P-JNK of the liver tissue and the body weight,urinary and drinking volume,glucose and lipid metabolism,insulin sensitivity index in these model rats.2.HepG2 cells were cultured in vitro to detect the effects of different concentrations of JieduTonluotiaogan Decoction?12.5ug/mL,25ug/m L,50ug/mL,100ug/mL,150ug/mL?on cell survival rate;The endoplasmic reticulum stress model of HepG2 cells were stimulated by 5 ug/mL tunicamycin for 24 hours,then detect the expression of IL-6,TNF-?,CRP,JNK2,IRE-1,NF-?B of these cells intervened by JieduTonluotiaogan Decoction.Results:1.The influences on the body weight,urinary and drinking volume:Both of the western medicine group and JieduTonluotiaogan Decoction group were declined on the quality of the body weight,but were still significantly different compared with the normal ZL rats;The urinary and drinking volume were significantly decreased,which were consistent with the therapeutic effect of JieduTonluotiaogan Decoction on T2DM patients.2.The influences on the glucose and lipid metabolism:The high concentration of JieduTonluotiaogan Decoction coupled with the western medicine group can significantly decline the blood glucose and improve the ISI of diabetic rats compared with the model group?^?P<0.05?,which proved the therapeutic effect on the diabetic rats in declining blood glucose,improving the insulin sensitivity and ameliorating the resistance;whereas the Glycosylated hematopoietic protein and INS have no statistical difference,which were due to the shorter duration.The high,medium and low concentration of JieduTonluotiaogan Decoction coupled with the western medicine group can significantly decline TC HDL-C and the high concentration of JieduTonluotiaogan Decoction can decline TG significantly(^?P<0.01,^??P<0.05),whereas compared with the western medicine group,the high concentration of JieduTonluotiaogan Decoction declined the TG,TC and HDL-C more significantly.The result proved that JieduTonluotiaogan Decoction can better decline the blood lipid.3.The aspect of immunohistochemistry:There is no high-glucose,high-fat in the ZL group and the expression of IRE1,P-IRE1,JNK and P-JNK protein in hepatocellular plasmids can not be observed,while there is high-glucose and high-fat in the model group,western medicine group and JieduTonluotiaogan Decoction group,the expression of IRE1,P-IRE1,JNK and P-JNKin hepatocellular plasmids can also be observed.This is an evidence that the ZDF mice can be induced to form a diabetic model by administering Purina#5008 diet.The expression of IRE1,P-IRE1,JNK and P-JNK in hepatocellular plasmids of the western medicine group and JieduTonluotiaogan Decoction group were significantly declined compared with the model group,which can prove that JieduTonluotiaogan Decoction and Metformin can ameliorate the insulin resistance and reduce the apoptosis of?cell by declining the expression of IRE1,P-IRE1,JNK and P-JNK and suppressing the IREl-JNK signaling pathways.The expression of IRE1,P-IRE1,JNK and P-JNK in hepatocellular plasmids of JieduTonluotiaogan Decoction group?high and low concentration?were more significantly declined than the western medicine group,and the expression IRE1,P-IRE1,JNK and P-JNK were the most significantly in the medium group,which prove that JieduTonluotiaogan Decoction can better ameliorate the insulin resistance and reduce the apoptosis of?cell.4.JieduTonluotiaogan Decoction can increase the sensitivity of insulin by increasing the expression of IRS-1 and PI3K in liver tissue.5.On the aspect of vitro cell culture:There is less cytotoxicity in the JieduTonluotiaogan Decoction group?12.5ug/mL,25ug/mL,50ug/mL,100ug/mL,150ug/mL?for the HePG2 cells;There is a small amount of IL-6,TNF-?,CRP,JNK2,NF-?B and IRE-1 in HePG2 cells,the expression of these inflammatory factors and proteins were significantly increased in the model group compared with the normal group when HePG2 cells were stimulated by Tunicamycin,which can prove that the endoplasmic reticulum stress model of HepG2 cells can be induced by 5 ug/mL tunicamycin for 24 hours;by the way,the expression of IL-6,TNF-?and CRP in the JieduTonluotiaogan Decoction group?high,medium and low concentration?were declined than the model group especially for the 100ug/mL group,which can prove that JieduTonluotiaogan Decoction inhibit the expression of inflammatory factors of HepG2 cells to improve the endoplasmic reticulum stress state;On the aspect of protein,the expression of JNK2,NF-?B in the high and medium concentration and the expression of IRE-1 in the high,medium and low concentration of JieduTonluotiaogan Decoction were significantly reduced especially for the medium concentration group,which can prove that the expression of JNK2,NF-?B can be declined by the the high and medium concentration of JieduTonluotiaogan Decoction,and the expression of IRE1 can be declined by JieduTonluotiaogan Decoction.The results show that JieduTonluotiaogan Decoction acts as the membrane insulin sensitizer to reduce the expression of inflammatory factors and proteins in HepG2 cells.Conclusion:1.JieduTonluotiaogan Decoction can better ameliorate the disorder of glucose and lipid metabolism,improve the insulin sensitivity and reduce the urinary and drinkingvolume.2.JieduTonluotiaoganDecoctioncandeclinetheexpressionof IRE1,P-IRE1,JNK and P-JNK and suppressing the IREl-JNK signaling pathways and increase the expression of IRS-1 and PI3K in liver tissue..3.There is less cytotoxicity in the JieduTonluotiaogan Decoction group for the HePG2 cells.4..JieduTonluotiaogan Decoction can inhibit the expression of inflammatory factors of HepG2 cells to improve the endoplasmic reticulum stress state,the expression of JNK2,NF-?B can be declined by the the high and medium concentration of JieduTonluotiaogan Decoction.