Consumer Paste Transfer The Turbidimetry Reversal Of Obese Non-alcoholic Fatty Liver Model Of Rat Liver Fatty Degeneration Of The Experimental Study

Objective: Nonalcoholic fatty liver disease(NAFLD) is a clinical characterized by excessive fat degeneration, without alcohol over storing ingestion in liver hepatic of fatty cellular. With the improvement of the standard of living, more and more people are suffering nonalcoholic fatty liver (NAFL). It appears that obesity is concerned with the cause of NAFL. And there is a tendency of young. The incidence fatty liver is increasing and NAFLD patients are prone to liver sclerosis or liver failure. So it is necessary to find a new way on therapy of fatty liver. There aren't any ideal medications to prevent NAFL. Chinese traditional medicine(TCM) possesses some superiority on it. My tutor, professor Tong Xiao-lin realized in his clinical work that the obesity is the"comment etiological factor"of NAFL,T2DM,hypertension etc. The treatment of obesity and NAFL with the theory of"Gao"and"Zhuo"is the representation of preservation theory. There a good therapeutic effect of the treatment of NAFL with Xiao Gao Zhuan Zhuo Preparation. Using the model of NAFL recognized by the world and linking to the evolution of modern research of NAFL, we observed the body weight,abdomen fat,the liver index,hepatic tissue pathology,Liver function,blood lipid,FFA,MDA,SOD,TNF-αand Leptin in the liver homogenate,hepatic tissue expression of CYP4502E1,InsR and IRS-2 protein and mRNA by the technique of pathology, biochemistry and molecular biology in the research, to offer deep study of pathogenesis of NAFL.Methods:The research was divided into two steps.The fInsRt step to study the effect of Xiao Gao Zhuan Zhuo Preparation on NAFL(Experiment I):Forty clean graded male Wister rats, weight 180-220g, ware fed 1W normally, then were randomly divided into 5 groups, namely normal control group, model group, diet control group, rosiglitazone group, Xiao Gao Zhuan Zhuo group. The normal control group are fed with ordinary diet; the rest are fed with high fat diet (88%ordinary diet,14% lard,1% cholesterin) to replicate NAFL model of rats; After 8W, the diet of the normal control group and the model group don't change, and diet of the rest change to the ordinary food. At the same time, the rats of the rosiglitazone group are poured suspension of rosiglitazone(3mg/Kg.d, qid)into stomach. the rats of Xiao Gao Zhuan Zhuo group are poured suspension of Xiao Gao Zhuan Zhuo Preparation into stomach. At the end of twelve week of the experiment,all rats are fasted for 12h, then are drawed blood(2ml) throuogh jugular vein, centrifugalization(3000rpm,10m),then to obetain the blood serum, stored at -20℃.After 2 days, all the rats are experienced the Clamp experiment. After Clamp experiment, all the rats are killed. Amone the whole experiment, we observe common state of animal. We reckon liver index, detect the con tent of serum ALT,AST CHO,TG,HDL-C,LDL-C and FFA in the liver homogenate. Observe hepatic tissue pathology with electron microscope.The second step is to study the mechanism of treatment of NAFL with Xiao Gao Zhuan Zhuo Preparation. The method of replicate NAFL model of rats,to divid group and the information of usage medicine are the same as the experimentⅠ. To detect the blood glucose and insulin. To detect the content of MDA and SOD in the liver homogenate. To observe the protein expression of CYP2E1 by immunohistochemistry and the mRNA expression of CYP2E1,InsR and IRS-2 by PT-PCR.Result:The experiment I is to study the effect of Xiao Gao Zhuan Zhuo preparation on NAFL. According to the hepatic tissue pathology and the state of hepatic function of rats, we succeeded in replicating the models of rats with NAFL. Compared with the model group, the body weight,abdomen fat and the liver index of Xiao Gao Zhuan Zhuo group decrease significantly(P<0.01). The degree of hepatic tissue inflammation is improved significantly(P <0.01)and the level of serum ALT,AST,CHO,TG,LDL-C and FFA in the liver homogenate decrease markedly(P <0.01).The second step (Experiment II-V):⑴Compared with the model group, Xiao Gao Zhuan Zhuo preparation can decrease significantly the content of TNF-αand Leptin (P <0.01,P <0.05).⑵Compared with the model group, Xiao Gao Zhuan Zhuo preparation can decrease significantly the content of MDA and increase markedly the activity of the SOD(P <0.01). Immunohistochemistry experiments display that the positive area and the MOD numerus of CYP2E1 decrease significantly(P<0.01).RT-PCR experiments show that the expression of CYP2E1 mRNA decrease markedly(P <0.01).⑶Compared with the model group, Clamp experiment display that Xiao Gao Zhuan Zhuo preparation can increase significantly the glucose infusion rate(GIR)(P<0.01).The fasting insulin of the Xiao Gao Zhuan Zhuo group decrease significantly compared with the model group (P<0.01).RT-PCR show that the expression of InsR and IRS-2 mRNA increase markedly(P <0.01).Conclusion:1. Xiao Gao Zhuan Zhuo Preparation can decrease significantly the body weight,abdomen fat and the liver index. The results display that Xiao Gao Zhuan Zhuo Preparation has the function of weight reduction. That hints: to reduce the abdomen fat may be one of the mechanism of Xiao Gao Zhuan Zhuo Preparation in the treatment of nonalcoholic fatty liver.2. Xiao Gao Zhuan Zhuo Preparation have the function of reduction the fat deposition in the hepatocyte and improve the degree of hepatic tissue inflammation.3. Xiao Gao Zhuan Zhuo Preparation can improve the the liver function and decrease the blood lipid,FFA in the liver homogenate. That illustrate Xiao Gao Zhuan Zhuo Preparation have the function of liver protection and decreasing lipid. 4. Through decreasing the content of TNF-αand Leptin in the liver homogenate,Xiao Gao Zhuan Zhuo Preparation may indirectly lessen the action of lipid peroxidation; Through lessening the Leptin resistance, Xiao Gao Zhuan Zhuo Preparation may improve the insulin resistance.5. Xiao Gao Zhuan Zhuo Preparation can inhibit the expression of the CYP2E1 protein and lessen the action of lipid peroxidation, meanwhile, it can increase the ability of anti- lipid peroxidation.6. Xiao Gao Zhuan Zhuo Preparation can raise the GIR and decrease the insulin. In the aspect of insulin signal transduction, Xiao Gao Zhuan Zhuo Preparation affect insulin receptor and insulin receptor substrate-2, thereby ameliorating the insulin resistance.