Study The Effect Of Shenshao Oral Liquid On Liver Injury In Diabetic Rats By TLR4/MyD88 Pathway

Objectives To explore the effect and mechanism of Shenshao decoction on diabetesinduced liver injury rats through detecting serum biochemical indexes,histomorphology and the expression of TLR4,My D88,TRAF6,NF-KB and other inflammatory factors,by constructing a model of liver injury in rats with type 2 diabetes mellitus,were treated with different dose of Shenshao oral liquid.Methods 1 Grouping: Fifty specific pathogen free male Wistar rats(170-200g)were randomly divided into two groups: control group(N=10),experimental group(N=40).2Model of type 2 diabetic rat: After adaptive feeding for 1 weeks,the control group was fed with normal diet,and the experimental group was fed with high fat diet,until 1 months after the start of modeling.The experimental group were induced by intraperitoneal injection of Streptozotocin(25mg/kg)(concentration 1%,p H 4.5),the C group injected intraperitoneally the same dose of citrate buffer.Three successive fasting blood glucose?16.7mmol/L and polydipsia,polyuria,polyphagia appear,initially confirmed the successful model of type 2 diabetes.3Drug intervention:After the completion of the model,the experimental group were divided into four groups model group,Shenshao decoction low dose group,Shenshao decoction middle dose group,Shenshao decoction high dose group Shenshao decoction preparation was used for drug experiment.Shenshao decoction low dose group Shenshao decoction middle dose group,Shenshao decoction high dose group according to 4ml/kg.d,8ml/kg.d,12ml/kg.d intragastric administration last 10 weeks.4Materials and processing:After 10 weeks,all rats were sacrificed,blood samples and hepatic tissue were obtained.Using full automatic biochemical analytic method to measure the level of blood glucose(GLU),alanine aminotransferase(ALT),aspartate amino shift enzyme(AST),total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL).Determination of antioxidant and oxide index in liver tissue by standard test methods of spectrophotometer and enzyme.HE staining was used to observe the morphological changes of liver tissue.Masson staining was used to observe the expression of fiber in liver tissues.The ultrastructural changes of liver cells and organelles were observed by transmission electron microscopy.Using immunohistochemical staining and western blot technology to observe the expression of liver tissue of rats in each group were TLR4,Myd88,TRAF6,NF-?B,IL-6 and other inflammatory factors.Results 1 High fat diet feeding combined with small dose of STZ intraperitoneal injection successfully completed the construction of T2 MD animal model.2 Biochemical indicators showed that: Compared with control group,the blood glucose,ALT,AST,TC,TG,LDL,MDA were significantly higher in the model group(P < 0.05),Shenshao decoction in the treatment group,the biochemical indexes were improved,blood glucose,ALT,AST,TC,TG,LDL,MDA decreased significantly(P<0.05),the difference was statistically significant 3 HE staining results showed that: Compared with the control group(C)group,the model group(M)liver cells swelling,fat degeneration,a large number of inflammatory cells gathered,the hepatic lobules lose the integrity of the structure,the liver line arrangement disorder is not neat,liver sinus expansion.After the oral administration of ginseng and ginseng oral liquid,the hepatic lobules were recovered completely,some of them were arranged in order,and the changes of balloon like cells,hyaline degeneration and inflammatory infiltration were reduced.The results indicated that Shenshao decoction could improve the liver injury caused by diabetes mellitus 4.Masson staining: A large number of thick blue collagen fibers can be seen in the blood vessels,periportal and disse space of liver tissue in model group.In shenshao group: Shenshao decoction low dose group changes little,thick collagen fibers were still visible.Blue collagen fibers decreased and thinner in Shenshao decoction middle dose group.Only a few blue collagen fibers were observed in Shenshao decoction high dose group,which was close to the normal control group.5Immunohistochemical Staining: The TLR4 protein was mainly expressed in the cytoplasm of the liver cells,and it was brownish yellow granules.Compared with control group,the deposition of yellow particles in model group was significantly increased.In the Shenshao decoction low dose group,the deposition of yellow particles decreased,and the deposition of yellow particles in Shenshao decoction low dose group and Shenshao decoction high dose group was significantly reduced.Similar expression were observed in NF-?B,Myd88,TRAF6,IL-6.6 Western blot: Compared with control group the expression of TLR-4?NF-?B?Myd88?TRAF6?IL-6 protein significantly increased in model group(P <0.05).In group of Shenshao decoction: the expression of TLR4?IL-6?TRAF6 protein was lower in Shenshao decoction low dose group than that in model group(P<0.05),The expression of TLR4?IL-6?My D88?TRAF6 protein in Shenshao decoction middle dose group was significantly lower than that in model group(P< 0.05),The expression of TLR4 ? IL-6 ? My D88 ? TRAF6 ? NF-?B protein was obviously reduced in group group T-H(P<0.05).Conclusions 1 Inflammation may involved in the process of liver injury in type 2diabetes mellitus..2 The protective effect of Shenshao oral liquid on diabetic liver injury may be mediated by TLR4/My D88/RAF6 dependent signaling pathway.