| With the development of social economy and the transformation of the national way of life, The incidence of the diseases such as diabetes, atherosclerosis, metabolic syndrome increased rapidly caused by modern high calorie intake and less sports consumption way of life, while the key link of these diseases are insulin resistance(IR). Therefore, how to improve the IR has become a research focus in recent years. Clinical medicines to improve IR currently are mainly biguanides and the Thiazolidinediones(TZD), which is greatly limited in clinical use due to the adverse reaction。So it’s a matter to select insulin sensitizing medicine. Traditional Chinese medicine emphasizes on regulating the body’s endocrine and metabolic activity in overall, systemic and multi-target way, which effect is much better than the single effect of Western medicine. Jinlida particles contains 17 kinds of Chinese medicine such as ginseng, solomon’s seal, fuling,Radix Ophiopogonis, kuh-seng,its clinical test shows that it can effectively reduce blood glucose and improve the IR of type 2 diabetic patients, but the mechanism is not clear.Objective:In this study, the formation of dyslipidemia and hyperinsulinemia in high fat fed Apo E-/- mice, has successfully constructed animal model of insulin resistance associated with lipid metabolism disorder,which is similar to the human disease by interaction of environmental and genetic factors. The liver plays an important role in energy metabolism of the whole body, which is the center organ of the conversion of carbohydrate, lipid and protein. The early changes of liver metabolism are closely related to the occurrence and development of IR, obesity and type 2diabetes. In-depth study on the liver can provide an important theoretical basis for the further study of IR. This research observes the changes of liver related genes in the condition of IR induced by high-fa Apo E-/- mice, then intervene the process by different doses of the Jinlida, in order to explore Jinlida mechanism, expecting to provide the theoretical basis for the clinical improvement of IR.Methods:8 male C57BL/6J mice were set to normal control group(NF), given normal feed; 40 healthy male Apo E-/- mice were fed with high fat diet for 16 weeks divided into 5 groups randomly: model group(HF), pioglitazone group(BGLT), Jinlida high dose group(JLDH), Jinlida middle does group(JLDM), Jinlida low dose group(JLDL), according to the body weight of mice, drug delivery of groups were as follows :the high dose group 3.8 g/kg.d-1, the middle dose group 1.9 g/kg.d-1, the low dose group 0.95 g/kg.d-1, Pioglitazone Hydrochloride Tablets 1.33 mg/kg.d-1, drugs after porphyrization were suspended with pure water and set the volume to be 100 ml by gavage, See Table1. The normal control group, model group were given pure water, began gavage for 8 weeks. At the end of the experiment, glucose tolerance test(OGTT) to evaluate the degree of insulin resistance in mice, then blooding sampling in mice eyes, collecting serum, killing the mice, acquiring the liver specimens to preserve in liquid nitrogen standby.Determination of mouse serum fasting blood glucose(FBG), free fatty acid(FFA), cholesterol(TC), triglyceride(TG), high density lipoprotein cholesterol(HDL-C), low density lipoprotein cholesterol(LDL-C).To detect the level index of the fasting plasma insulin by radioimmunoassay(FIns) Analysis method, to calculate insulin sensitivity index(ISI), and evaluate the degree of insulin to resist.Liver HE staining was performed to observe the changes of liver tissue of mice and lipid deposition degree of mouse.Western blotting(Western Blot)determination of hepatic insulin receptors of mice(INSR), insulin receptor substrate 1(IRS-1), fatty acid translocase(FAT), carnitine acyltransferase 1(CPT1).Results:1 To improve the blood biochemical index in miceCompared with the control group, the fasting blood glucose(FBG), cholesterol(TC), triglyceride(TG), free fatty acid(FFA), low density lipoprotein cholesterol(LDL-C) of the model group mice were significantly increased(P<0.05), high density lipoprotein receptor(HDL-C) significantly decreased(P<0.05), These suggested that the animal models induced by high fat diet in this experiment is successful.Compared with the model group, Jinlida reduced the fasting blood glucose(FBG), cholesterol(TC), triglyceride(TG), free fatty acid(FFA), low density lipoprotein cholesterol receptor(LDL-C) of the experimental mice in different degrees, increased high density lipoprotein receptor(HDL-C)(P<0.05), and improves blood biochemical indexes of the mice.2 To improve insulin sensitivity in miceCompared with the control group, the fasting insulin(FIns) and insulin sensitivity index(ISI) of the model group mice were significantly increased(P<0.05),these explained that animal model of insulin resistance made in this experiment induced by high fat diet is successful.Jinlida each dose group can reduce the fasting insulin(FIns) level, and the insulin sensitivity index(ISI). In the oral glucose tolerance test, each mouse all appear that blood sugar levels rose to the peak, then all was gradually decreased; compared with the control group, Each time point of blood glucose level of the model group were significantly increased; Jinlida drug group compared with model group, blood glucose peak and shortening the time of blood glucose were decreased significantly, Jinlida drug group had significant difference compared with the model group(P<0.05).3 Improvement of mouse liver steatosisUsing HE staining,Jinlida was found it can alleviate the changes of Liver fatty degeneration in mice induced by high fat to varying degrees.4 Effect on the expression of lipid metabolism related proteins in mouse liverIt was found that each dose group of Jinlida increased the expression of insulin receptor and insulin receptor substrate moleculesto varying degrees.5 Effect on the expression of lipid metabolism related enzymes in mouse liverEach dose group of Jinlida can increase expression of carnitine palmitoyltransferase 1 in different degree, reduce the expression of fatty acid translocase molecules.Conclusion:Jinlida can improve the blood biochemical index, improve insulin sensitivity, reduce liver fat disease, and improve the expression of hepatic lipid metabolism related proteins and enzymes in model mouse. Therefore, the mechanism of Jinlida improving IR of the mice liver may be: Jinlida can lower the expression of FAT/CD36, raise the expression of CPT1, on the one hand it can reduce the intake of long-chain FFA, on the other hand it can promote the beta oxidation of fatty acids in the liver, and then promote the oxidation of fatty acids and reduce lipid deposition in the liver to improve IR. |
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