The Study On The Mechanism Of Wendan Decoction And Jiawei Wendan Decoction For Insulin Receptor Singnaling Pathways In IGT Rat

Objectives:The experimental study on normal male SD with feeding high fat diet for 20 weeks,which is close to the onset of human IGT process,to establish a simple diet induced type IGT animal models.The pathogenesis of IGT was discussed through the signal pathway of insulin receptor,to further study mechanism and effect target of IGT rats fed with Wendan Tang and Jiawei Wendan Tang,provide experimental basis for clinical treatment of IGT,and provide technical platform for replication rat model of IGT.Methods:Rats uping to the standard of IGT model were randomly divided into Wendan Tang group(WDT),Jiawei Wendan group(JWWDT)and IGT model group(IGT),Ten rats feeding 20 weeks of ordinary food were set to normal control group(NC).NC and IGT were given distilled water to fill the stomach with 10 ml/kg volume in a fixed time every day,WDT(6.18 g/kg)and JYWDT(11.25 g/kg)was given by intragastric administration of the same volume of corresponding compound water decoction.Observing the medical morphology of liver,skeletal muscle and fat with staining by HE.The expression of the PI3K,PKB,GSK-3 beta and GLUT-4 protein was determined by immunohistochemical method in rat liver,skeletal muscle and fat tissue.Expression of the five phosphorylated proteins and total protein(PKB,GSK-3 beta,GLUT-4,p38,ERK1/2)in liver and skeletal muscle was determined by Western Blot method.The three mRNA(GLUT-4,ERK1/2,p38 MAPK)was measured by RT-PCR method.The experimental study is to explore the pathogenesis of IGT,targets of Wendan Tang and Huayu Wendan Tang.Results:(1)Liver tissue:We can see clearly the nucleus and cytoplasm from NC liver cells,cell morphology is in good condition and liver plate arrangement neat.IGT rats appeared the nucleus pycnosis,uneven cytoplasmic staining,fatty degeneration,with a small amount of inflammatory cells.Compared with IGT,liver cells form of medication group is neat,some liver sinus is clearly visible,there are small pyknotic nucleus.Skeletal muscle tissue:skeletal muscle bundle of the NC rats rows neatly,the nucleus is located in the muscle on the edge of the beam with a flat,no inflammatory cells exist.Uneven dyeing of IGT rats skeletal muscle cell appears,muscle fiber arrangement is out of order,Eosinophilic cells and the nucleus pycnosis exist.Muscle fiber of medication group arrangement is neater,the nucleus pycnosis is less than IGT,and muscle fibers ranked neatly.Fat tissue:The NC fat cell morphology is perfect,clear and neat.Fat cells of IGT rats is dyed unevenly,cell shape is irregular,the nucleus pycnosis existed,capillary significantly increased,there are scattered in the red blood cells.Compared with IGT,The cell membrane of medication group is relatively clear and neat,the nucleus pycnosis is less,capillary significantly reduced,a few red blood cells appeared.(2)Compared with NC,the PI3K PKB,GLUT-4 protein expression of IGT rat skeletal muscle decreased significantly(P<0.01 or P<0.05),GSK-3 beta protein expression was significantly increased(P<0.01 or P<0.05),WDT and JWWDT all can be a significant rise with PI3K,PKB,GLUT-4 protein expression in rat skeletal muscle(P<0.01 or P<0.05),reduce the GSK-3 beta protein expression(P<0.01 or P<0.05);Compared with NC,PI3K,GLUT-4 protein expression of IGT rats in liver tissue decreased significantly(P<0.01 or P<0.05),PKB and GSK-3 beta protein expression significantly increased(P<0.01 or P<0.05),PI3K,GLUT-4 protein expression of WDT and JWWDT can be a significant rise in rat liver(P<0.01 or P<0.05),the PKB protein expression significantly reduce(P<0.01 or P<0.05),GSK-3 beta protein expression without adjustment.(3)Compared with NC,ERK1/2,p38 MAPK protein expression in IGT rat skeletal muscle,decreased significantly(P<0.01 or P<0.05),ERK1/2,p38 MAPK protein expression of WDT and JWWDT,all can be a significant rise(P<0.01 or P<0.05);Compared with NC,ERK1/2 protein expression in IGT rat liver,there were no significant difference,p38 MAPK protein expression was significantly higher(P<0.01);WDT can significantly reduce ERK1/2,p38 MAPK protein expression(P<0.01),but JWWDT significantly elevated ERK1/2 and p38 MAPK protein expression(P<0.01).(4)Compared with NC,ERK1/2,p38 MAPK,GLUT-4 mRNA expression in IGT rat skeletal muscle,was significantly increased(P<0.01),while WDT,JWWDT can significantly reduce ERK1/2,p38 MAPK,GLUT-4 mRNA expression(P<0.01);Compared with NC,ERK1/2,p38 MAPK.GLUT-4 mRNA expression in IGT rat liver significantly reduced(P<0.01).while WDT,JWWDT all can be a significant rise of ERK1/2,GLUT-4 mRNA expression(P<0.01 or P<0.05),JWWDT can also be a significant rise of p38 MAPK mRNA expression(P<0.05).ConcIusions:Long-term high fat diet can build up IGT animal model in rats similarities to the standard of human,characterized by insulin receptor signal pathway transduction disorder,such as abnormal expression of protein of PI3K and MAPK pathway.Wendan Tang and Jiawei Wendan Tang can significantly adjust its abnormal expression.but it is different of the targets and the adjustment of protein for two prescriptions.