| BackgroundInsulin resistance is the common pathological and physiological basis of various diseases, such as type 2 diabetes, hyperglycemia, obesity, hyperlipidemia, coronary heart disease and so on. Improving insulin sensitivity and improving insulin resistance is an important task for the prevention and treatment of endocrine diseases such as type 2 diabetes, hyperlipidemia and obesity. Based on years of clinical experience, a large number of literature review, Professor Gao Sihua proposed the regulation the function of liver, spleen and kidney in treatment based on syndrome differentiation of type 2 diabetes. With nourishing qi and yin of spleen, liver and kidney, accompanied by heat treatment and activating blood circulation, He has created the clinical prescription of Jiangtang Xiaoke granules (JTXKG). Preliminary experimental and clinical observations have confirmed the effectiveness of the prescription in correcting glucose and lipid metabolism. Since the KK-Ay type diabetic mice for their fur color mutation of the Ay gene, with genetic susceptibility and environmental sensitivity, through diet induced spontaneous emergence of high blood sugar, obesity, hyperinsulinemia, endocrine disorders are more likely to be induced, emergence of metabolic syndrome. As a model of obese type 2 diabetes mellitus, the typical insulin resistance is the pathological change, which is the most close to human type 2 diabetes mouse model. Skeletal muscle, as the largest target organ of insulin, is responsible for more than 70%of the glucose metabolism. Therefore, we choose KK-Ay mouse skeletal muscle as the research medium, we conducted a deep study on the mechanism of the hypoglycemic mechanism.Research objectiveWith the observation effects of obesity weight, glucose and lipid metabolism and PI-3K pathway in skeletal muscle of Jiangtang Xiaoke granules in type 2 diabetic model KK-Ay mice. We investigate the possible mechanism of Jiangtang Xiaoke granules in the regulation of glucose and lipid metabolism.Research methodsThe 8 week old KK-Ay mice fed with high fat diet, at their 12 weeks of age,50 mice, whose fasting blood glucose were not less than 14mmol/l, were randomly divided into 5 group (control, JTXK hign, JTXK medium, JTXK low and pioglitazone). All mice were administered for next 10 weeks. Food intake, body weight and fasting blood glucose were recorded every week. At week 4 and week 10 the glucose tolerance was tested. At the end of the experiment, blood glucose, serum insulin, hemoglobin, blood lipids and free fatty acid levels of 4 and free fatty acids were detected, detection of insulin signal transduction pathway of PI-3K pathway in IRS-1, PI3K, AKT, GLUT4, GSK-3 beta RNA water with genes related to real time fluorescence quantitative PCR, and detection of protein expression of the above mentioned factors in skeletal muscle by Blot WesternResearch resultsEach dose of JTXK had reduced food intake in KK-Ay mice, The body weights of all mice were continuing to rise and high-dose JTXK administration could significantly begin to reduce the body weight gain from 8th week. JTXK lowered fasting blood glucose after 4th week administration and medium dose was most effective in long-term. JTXK lowered the level of glycosylated hemoglobin, and the medium-dose was most effective. JTXK could improve insulin sensitivity, and high-dose was most effective. Glucose tolerance was not changed at week 4, but showed a significant improvement in JTXK high group at week 10. serum TG, TC, HDL-C and LDL-C significantly improved, with a low dose of FFA levels Improved by JTXKReal time fluorescence quantitative PCR results showed that JTXK has a rising trend to IRS-1, PI3K, Akt and GLUT4 expression, reduced GSK-3 beta mRNA, in which the effect of the high-dose group is particularly significant, and similar with the positive drug pioglitazone effect. Western blot results showed that at the protein level, in KK-Ay mouse skeletal muscle tissue,the expression of PI3K, Akt, p-Akt, GLUT4 protein increased in all, of GSK-3β is decreased, which the effect of high dose is the most obvious.ConclusionJTXK promote KK-Ay mice weight loss and metabolic, may inhibit food intake of mice。 controlled FBG to and improve glucose tolerance, reduce HDL-C, LDL-C, TG, TC and FFA levels in Long term administrationJTXK increased the expression of IRS-1 mRNA,PI3K and Akt mRNA and GLUT4 mRNA, reduce GSK-3 beta mRNA expression in KK-Ay mice skeletal muscle.JTXK had promoted expression of IRS-1,PI3K, Akt, p-Akt, GLUT4 protein, reduce GSK-3β protein expression in KK-Ay mouse skeletal muscle tissue to improve insulin resistance.The innovation of this study:It is for the first time and in the traditional Chinese medicine theory to discusse the close relationship between the dysfunction of Liver, Spleen and Kidney and insulin resistance, and reflect the important significance in Liver, Spleen and Kidney for the treatment of type 2 diabetes from the correct point of view about insulin resistance in skeletal muscle.It is for the first use of Jiangtang Xiaoke granules to intervent of spontaneous diabetes mellitus model KK ay mice in order to observe its Pharmacological effects on obesity type 2 diabetic mice model, and to reveal the effect of the treatment of Liver, Spleen and Kidney on obesity type 2 diabetic blood glucose and lipid metabolic disorder.It is for the first time, the TCM theory and insulin signal transduction of PI3K pathway has be linked up, and application of KK-Ay spontaneous diabetic mice model, and to observed glucose and lipid metabolism and other aspects of the role of pharmacy of JTXK which is based on the prescription of theory of regulating the function of liver, spleen and kidney together. And on the basis, through the research of JTXK on insulin signaling pathway-PI3K pathway key factors in gene and protein influence, to investigate the effects of JTXK on PI3K signal transduction pathway, and reveal its possible mechanism of action, and provide of PI3 K Signal transduction pathway which is one of the most important pathway in glucose and lipid metabolism, so as to reveal the possible working mechanisms of JTXKG and provide the experimental data proof for the novel theory in insulin resistant treatment.It is for the first time with different doses of Jiangtang Xiaoke granules to intervent type 2 diabetic mice model,and we found that the high dose group is the most significant to body weight and insulin resistance, and middle dose group is most effective to fasting blood glucose and hemoglobin levels, and low dose group is most effective to reduce the free fatty acids,and that means Jiangtang Xiaoke granules does not exist in a dose-dependent relationship in the treatment process. |
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