| Research purposes: To establish a mouse model of type 2 diabetes(T2DM)by intraperitoneal injection of high-fat feed combined with alloxan(ALX),and on the basis of the treatment of T2 DM model mice Pharmacodynamic evaluation research provides scientific basis for the treatment of T2 DM by this prescription.Research method: 66 SPF Kunming male mice were randomly assigned to 6 mice as a normal control group and fed with basic feed.The remaining 60 mice were fed with high-fat diet for 4 weeks,and the model mice were given once every other day,and three consecutive times of intraperitoneal injection of ALX120mg/kg(PH=4.4)alloxan(Alloxan)to establish type 2 diabetic mice In the model,on the 3rd day after the last injection,the tail blood collection was performed to detect the level of basal fasting blood glucose [(FBG)12-hour fasting but not water],and FBG≥ 11.1mmol/L was used as a sign of the success of type 2 diabetes model.At the same time,it was divided into a normal group(N=6)and a model group(N=12)(intragastric administration with normal saline: 1.5ml/10g),the Mongolian medicine Tegoshidegeqi high-dose group [(N=12)3g/kg ],Mongolian medicine Tegshidgeqi medium-dose group [(N=12)1.5g/kg],Mongolian medicine Tegshidgeqi low-dose group [(N=12)0.75g/kg] and metformin hydrochloride group [(N=12)200mg/kg],a total of 6 groups.The mice in each group were administered continuously for 21 days(three weeks).During the administration period,except for the normal group,the other groups were fed with high-fat feed.The tail vein blood was collected every week to detect the fasting blood glucose(FBG)and body weight of the mice in each group,and the status of the mice in each group was observed and recorded.After the experiment,the FBG level and body weight were measured,then the eyeballs were taken for blood collection,and the neck was sacrificed.The pancreas,liver,kidney,skeletal muscle and adipose tissue of the mice were taken to detect relevant indicators.Detect the levels of cholesterol(TC),triglyceride(TG),highdensity lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)in the serum of mice with an automatic biochemical analyzer HE staining and pathological analysis,ELISA method was used to detect the levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and serum insulin(FINS)in the serum of each group of mice and calculate the insulin resistance index(HOMA-IR),kit method for determination of superoxide dismutase(SOD)activity and reduced glutathione(GSH)content in mice liver and kidney(left side)tissue antioxidant capacity,Western blot(Western Blot))Method to detect the expression of GLUT-4 protein in skeletal muscle(left hind gastrocnemius muscle)and fat(abdominal omentum fat)tissues of mice in each group.Research result:1.General situation of mice in each group:(1)After 4 weeks of feeding with high-fat diet,the body weight of the mice increased significantly,and the mouse model of hyperlipidemia was successfully established.Then,once every other day,three consecutive intraperitoneal injections successfully established the mouse model of type 2 diabetes.After modeling,the average fasting blood glucose of T2 DM model mice was 15.785 mmol/L.(2)Mice in type 2 diabetes model group have poor mental state,rough hair,yellow color,and water intake Diabetes symptoms such as increased,increased food intake,and significantly increased urine output are obvious,and the new Mongolian medicine Tegexi Degeqi(Danureux)powder can relieve the above symptoms in model mice.2.Comparison of fasting blood glucose(FBG)levels of mice in each group: The results showed that before the administration,the FBG of the model group,positive drug group,Mongolian medicine Tegshidgeqi high-dose group,middle-dose group,and low-dose group was higher than that of the normal group(P<0.05),indicating that type 2 diabetes was less The mouse model was successfully established.Three weeks after administration,FBG in the positive drug group,high-dose group,medium-dose group,and low-dose group was lower than that in the model group(P<0.05),indicating that the drug had a hypoglycemic effect.Compared with the high-dose group and the low-dose group,the Mongolian medicine Tegshidgeqi medium-dose group had a higher FBG decrease(P<0.05),indicating that the Mongolian medicine Tegshidgeqi middle-dose group had the best effect.3.Comparison of blood lipids of mice in each group: The TC,TG and LDL-C values of the model group were higher than those of the normal group(P<0.05),the HDL-C values of the model group were lower than that of the normal group(P<0.05),and the high-dose group and medium dose of Mongolian medicine Tegheidegeqi The TC,TG and LDL-C values of the dose group and the low dose group were lower than those of the model group(P<0.05),and the HDL-C value ratio model of the Mongolian medicine Tegshidgeqi high dose group,middle dose group and low dose group Group high(P<0.05),indicating that the drug has the effect of inhibiting the increase of TC,TG,LDL-C value and HDL-C value.Compared with the high-dose group and the low-dose group,the medium-dose group of Mongolian medicine Tegshidgeqi had a higher increase in HDL-C value(P<0.05),indicating that the medium-dose group had the best effect in raising HDL-C.Compared with the low-dose group,the middle-dose group of the Mongolian medicine Tegshidgeqi had a greater decrease in TC value(P<0.05),indicating that the middle-dose group had a better effect on reducing TC than the low-dose group.The three different doses of Mongolian medicine group had no difference in the decrease of TG and LDL-C values(P>0.05).4.Comparison of anti-inflammatory effects of mice in each group: The results showed that the serum IL-6 and TNF-α in the model group were higher than those in the normal group(P<0.05),and the serum IL-6 and TNF-in the high-dose group,medium-dose group and low-dose group α is lower than the model group(P<0.05),indicating that the drug has the effect of reducing serum IL-6 and TNF-α.Compared with the high-dose group and the low-dose group,the medium-dose group of Mongolian medicine Tegshidgeqi had a higher decrease in IL-6 and TNF-α(P<0.05),indicating that the medium-dose group had the best effect.5.Comparison of serum insulin(FINS)levels of mice in each group: The FINS level of the model group was higher than that of the normal group(P<0.05),and the FINS level of the positive drug group,Mongolian medicine high-dose group,middle-dose group and low-dose group was higher than the model group(P<0.05),It indicates that the drug has the effect of improving hyperinsulinemia in type 2 diabetes model mice6.Comparison of antioxidant capacity of mice in each group: The results show that the liver and kidney SOD activity and liver and kidney GSH content of the model group are lower than that of the normal group(P<0.05),the Mongolian medicine Tegshidgeqi high-dose group,middle-dose group,low-dose group liver and kidney SOD,liver and kidney GSH is higher than the model group(P<0.05),indicating that the drug has elevated liver,Kidney SOD and liver and kidney GSH.Compared with the high-dose group and the low-dose group,the middle-dose group of Mongolian medicine Tegheedgeqi had higher levels of liver and kidney SOD and liver and kidney GSH(P<0.05),indicating that the middle-dose group had the best effect.7.HE stained pathological section: The pathological changes of pancreas tissue in the intervention group of Mongolian medicine Tegeshidegeqisan in various doses were significantly reduced compared with the model mouse group.8.Comparison of the expression levels of GLUT-4 protein of mice in each group: Western Blot was used to examine the expression level of GLUT-4 protein in skeletal muscle and adipose tissue.The Mongolian medicine Teguedge Qisan powder intervention group can promote the expression and translocation of GLUT-4 in skeletal muscle and adipose tissue,thereby increasing Glucose absorption increased significantly compared with the model group.9.Comparison of mouse insulin resistance index(HOMA-IR)in each group: The results showed that the HOMA-IR value of the model group was higher than that of the normal group(P<0.05),and the Mongolian medicine Tegshidgeqi medium-dose group had a higher decrease compared with the high-dose group and the low-dose group(P<0.05).It shows that the Mongolian medicine Teghidge Qisan has the effect of reducing the HOMA-IR value.Analysis conclusion:1.Continuous feeding of high-fat feed and intraperitoneal injection of alloxan succeeded in the successful preparation of a mouse model of type 2 diabetes.This model is suitable for the study of type 2 diabetes,and the modeling success rate reached 66.7%(FBG = 15.785 mmol/L).2.The new Mongolian medicine Tegexi Degeqi(Danurea)powder reduces blood glucose,inflammatory factors,insulin levels and insulin resistance index in mice with type 2 diabetes,and improves lipid metabolism.3.The Mongolian medicine Tegeshidege Qisan can reduce the damage of pancreatic islets of type 2 diabetes model mice and reduce the oxidative stress state of liver and kidney tissues.4.The Mongolian medicine Tegeshidege Qisan increases the protein expression of GLUT-4 in skeletal muscle and adipose tissue of type 2 diabetes model mice,and improves insulin resistance in type 2 diabetes mice. |
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