Establishment Of An Animal Model Of Type 2 Diabetes Mellitus With Both Qi And Yin Deficiency

Objective: Three methods were used to establish a rat model of type 2 diabetes mellitus with both Qi and Yin deficiency.According to the clinical diagnosis and treatment standards,the macro signs and micro indicators of the animal model were comprehensively collected,and the animal models with short modeling cycle and stable model were screened.The potential biological characteristics of the syndrome were discussed by metabonomics.Methods: High sugar and high fat + injection of streptozotocin(STZ),high sugar and high fat + injection of STZ + served hot Chinese traditional medicine and spontaneity GK rats were used to established long-term preparation of qi and Yin deficiency type rats model combined disease and type 2 diabetes mellitus.The mental state,body weight,food intake,water intake,grip,heart rate,respiration,pulse,tongue image and open field of the rats were monitored regularly.At the end of the experiment,the rat organ index,insulin resistance index(IR),random blood glucose,fasting blood glucose,four items of coagulation,four items of blood lipid,insulin(INS),SOD,MDA,c AMP,c GMP,CD4,CD8,a disintegrin and metalloproteinase with thrombospondin motif 4(Adamts4),early growth response 2(Egr2),insulin-like growth factor binding protein-1(Igfbp1)and urinary protein.HE staining was used to observe the histopathologic changes of pancreas,heart and kidney.The syndrome manifestations of type 2 diabetes rats with Qi-Yin deficiency were comprehensively evaluated.Metabolomics was used to screen the differential metabolites and metabolic pathways related to Qi-yin deficiency syndrome in rat urine.Results:(1)Three model rats showed symptoms of lethargy,emaciation,thirst,hyperphagia,fatigue,shortness of breath and weak pulse.○1 The level of random blood glucose,fasting blood glucose,insulin resistance index,TC,TG,LDL-C increased,HDL-C did not change significantly,FIB increased,APTT decreased,INS decreased,MDA increased,SOD did not change significantly,CD4 increased,CD8 did not change significantly,c AMP did not change significantly,c GMP decreased,Adamts4,Egr2,Igfbp1 and urinary protein increased in high sugar and fat +STZ group.○2 Random blood glucose,fasting blood glucose,insulin resistance index and TG increased,TC,LDL-C,HDL-C did not change significantly in rats with high sugar and high fat +STZ+ hot TCM model.FIB increased,APTT decreased,INS decreased,MDA increased,SOD decreased,CD4 increased,CD8 had no significant change,c AMP had no significant change,c GMP decreased,Adamts4,Egr2,Igfbp1 had no significant change and urinary protein increased.○3 The level of random blood glucose,fasting blood glucose,TC and HDL-C was increased,and TG was decreased.There were no significant changes in LDL-C.The level of PT,FIB and MDA increased,SOD increased,,CD4 increased,CD8 decreased.Adamts4,Egr2,Igfbp1 did not change significantly.urinary protein increased in spontaneous GK group.(2)Metabolomics results showed that ○1 The top 20 differential metabolites of high sugar and high fat + STZ model rats included threonine,leucine,lysine,valine,aspartic acid,etc.○2 The top 20 differential metabolites of high sugar and high fat +STZ+ hot Chinese medicine model rats included 3-isoallyl glutaric acid,norlichexanthone,tyrosine,serine,valine,phenylalanine,histidine,etc.○3 The top 20 differential metabolites of spontaneous GK model rats included L-lysine,L-phenylalanine,arginine,tryptophan,D-Glucaro-1,4-lactone,hypoxanthine,etc.Conclusion:(1)The three models can all have significant changes in macro signs,glucose and lipid metabolism,inflammatory factors,oxidative stress response and other aspects,showing the syndrome of Qi and Yin deficiency.But all the indicators have varying degrees of change.The experimental period of GK rats with spontaneous hyperglycemia is short and the of complications are more serious.It is expensive,which is not suitable for widespread popularization.The high sugar and high fat +STZ+ hot TCM model takes a relatively short time to build relatively,and the animals used are SPF grade.The cost is lower,and the reflected disease characteristics and syndrome characteristics are more consistent with the clinic.Therefore,it is more suitable for the preparation of the animal model of type 2 diabetes syndrome combined with Qi and Yin deficiency in this study.(2)Metabolomics of urine of three model animals showed that tryptophan,glutamic acid,threonine,arginine,glycine,lecithin,palmitic acid,lactic acid,5-hydroxyvalproic acid were potential differential metabolites of Qi and Yin deficiency.The metabolic pathways involved are amino acid biosynthesis,fatty acid biosynthesis,glycolysis/gluconeogenesis,drug metabolism-cytochrome P450 metabolism,etc.