The Efeect Of Jinlida On Lipid Metabolism In Skeletal Muscle Of Fat-Induced Insulin Resistance APOE^-/- Mice

ObjectiveThe main content of this topic research is hypoglycemic Chinese medicine compound Jinlida, it is a proprietary Chinese medicine,composed of ginseng,rhizoma polygonati,rhizoma atractylodis,bitter ginseng,radix ophiopogonis,radix rehmanniae,etc more than ten of TCM,but it is unclear of the mechanism in the specific hypoglycemic. Clinical studies have found that it can reduce the patient’s cholesterol,triglycerides,this study was to explore the Jinlida to put by adjusting the skeletal muscle lipid metabolism improving the mechanism of action of insulin resistance (IR).MethodsThis research is divided into the following three parts:1 Jinlida can improve the ApoE-/- mice induced by high-fat in skeletal muscle of insulin resistanceAutomatic biochemistry analyzer is used to detect the serum of blood glucose FBG,cholesterol,triglyceride TG,free fatty acid,high-density lipoprotein cholesterol,low density lipoprotein cholesterol level in mice.Fasting insulin levels was measured by immunoradiometric analysis,calculating indexes of insulin resistance and impaired glucose tolerance test was to evaluate the degree of insulin resistance.Skeletal muscle in mice hematoxylin eosin staining,oil red O staining and transmission electron microscope were used to watch the mice in skeletal muscle of tissue changes and lipid levels.The protein expression of insulin receptor, insulin receptor substrate 1,insulin receptor substrate 2 and glucose transporter 4 in skeletal muscle were measured by real-time quantitative reverse transcription PCR and Western blotting.2 The effect of Jinlida on lipid metabolic intermediate of cholesterol, triglycerides and fatty acids in skeletal muscle of fat-induced insulin resistance Apoe-/- miceExperimental groups and administered are same as the first part.The expression of the Cholesterol metabolism in skeletal muscle about the enzymes and gene:the protein expression of Low density lipoprotein receptor,sterol regulating element binding protein pyrolysis activated protein in skeletal muscle were measured by real-time quantitative reverse transcription PCR and Western blotting.The expression of the triglycerides in skeletal muscle about the enzymes and gene:TG content in skeletal muscle was measured by enzymic enzymatic.The protein expression of hormone sensitive lipase, fatty triglycerides enzyme, peroxidase body growth activated receptor gamma in skeletal muscle were measured by real-time quantitative reverse transcription PCR and Western blotting.The expression of the fatty acids in skeletal muscle about the enzymes and gene:the kit on skeletal muscle FFA content is determined using tissue FFA.The protein expression of carnitine acyltransferase 1,peroxidase body growth activated receptor alpha,fatty acid transposition enzyme in skeletal muscle were measured by real-time quantitative reverse transcription PCR and Western blotting.3 The effect of Jinlida on MAPK signaling pathways in skeletal muscle of fat-induced insulin resistance in Apoe-/- miceThe protein expression of mitogen activated protein kinase signaling pathway of extracellular signal regulated kinase and phosphorylated extracellular signal regulating kinase,P38 lightning,p-P38 lightning, stress activated protein kinase,phosphorylation stress activated protein kinase in skeletal muscle were measured by Western blotting.Results1 Jinlida can improve the ApoE-/- mice induced by high-fat in skeletal muscle of insulin resistanceImprove the blood biochemical indexes in mice:HF group compared with the NF group FBG, TC, TG, FFA and LDL-C levels were significantly increased,HDL-C levels drop;Compared with HF group tianjin eed groups can reduce mice different level of FBG, TC, TG, FFA and LDL-C, raising HDL-C,regulate blood sugar, blood lipid in mice.Improve insulin sensitivity in mice:HF group compared with the NF group FIns, HOMA-IR,glucose tolerance in mice the blood glucose levels were significantly elevated of each point in time.Compared with HF group Jinlida is able to cut different level FIns,lower HOMA-IR,to varying degrees of inhibiting glucose area under the curve of the increase,and presents a certain dose-response relationship.Improve the mice skeletal muscle tissue pathology:compared with NF group,HF group SCAP higher expression;Jinlida is able to reduce high-fat induced mice of different level of skeletal muscle lipid,improve mice skeletal muscle wire arrangement and the structure of mitochondria.Improve mice skeletal muscle insulin signal transduction:compared with NF group,HF group INSR, IRS-1, IRS-2 and GLUT4 mRNA and protein levels significantly decreased;JLDM,JLDH group raised the INSR,IRS-1,IRS-2 and GLUT4 mRNA and protein levels significantly.2 The effect of Jinlida on lipid metabolic intermediate of cholesterol, triglycerides and fatty acids in skeletal muscle of fat-induced insulin resistance Apoe-/- miceRegulate the expression of skeletal muscle cholesterol metabolism related genes and enzymes:compared with NF group,HF group SCAP higher mRNA and protein expression, reduced LDLR mRNA and protein expression.JLDM,JLDH dose group can cut SCAP mRNA and protein level,at the same time higher LDLR mRNA and protein expression.Regulate the expression of skeletal muscle triglyceride metabolism related genes and enzymes:compared with NF group,HF group of skeletal muscle an HSL,ATGL,PPAR gamma mRNA and protein levels were significantly higher than that of NF group;Compared with HF group,JLDM,an HSL JLDH group,ATGL,PPAR gamma mRNA and protein level to some extent.Influence of skeletal muscle lipid metabolism related genes in mice,enzyme expression:compared the NF group,HF group FAT/CD36 higher mRNA and protein expression,CPT1, PPAR alpha mRNA and protein expression;JLDM, JLDH dose group can cut FA17CD36,different level raised CPT1,PPAR alpha mRNA and protein expression.3 The effect of Jinlida on MAPK signaling pathways in skeletal muscle of fatinduced insulin resistance Apoe-/- miceThe Effect of the MAPK signaling pathways:compared with NF,HF group p-JNK, p-ERK,p-rising P38 lightning expression;Compared with HF group,JLDH can inhibit p-phosphorylated P38 lightning.Conclusion1 Jinlida is able to adjust the level of blood lipid parameters of insulin resistance in mice,improve insulin sensitivity, reduce the mice skeletal muscle lipid,improving the structure of the organelles in mice,mice can raise different levels of INSR,IRS-1, IRS-2 and GLUT4 protein and gene level,improve the insulin resistance in mice from different aspects.2 Jinlida improve insulin resistance in mice regulating from cholesterol,triglycerides and fatty acids of related enzymes.3 Jinlida can through a certain degree of inhibition of phosphorylated P38 lightning,blocking the MAPK signaling pathway to improve skeletal muscle insulin resistance in mice.