Study On The New Type Incretin And Insulin-mimetic Medicine For Diabetes Mellitus

Diabetes mellitus(DM)reputed as the most challenging health problem in the world.DM is a multi-level,multi-factor metabolic disease.Type 2 diabetes mellitus(T2DM)account for 90-95% of the total DM patients.T2 DM will lead various complications and the cardiovascular diseases,which are the major cause of death and disability in DM patients.Therefore,it urgent to development new type drugs to prevent and cure DM and its complications.In present study,two new type incretin and one insulin-mimetic drugs for T2 DM were constructed and systematically studied.The results summarized as follows.(1)By using designing of sgRNA and specific primers,recombinant PCR,a series intermediate vectors,the engineering agrobacterium EHA105＿NK contained p X6-U6-5rol GLP-1-HV donor vectors and p ZHY988-Cas6-sgRNA editing vectors were successfully constructed.Using the engineering agrobacterium EHA105＿NK to transform cucumbers,the hypoglycemic anti-thrombus bifunctional cucumber with the incretin-hirudin gene(5rol GLP-1-HV)knock-in the Bt target site was obtained,and the therapeutic effect of which was confirmed by mice model.(2)The sgRNA expression vectors p YES2-g RNA targeting S.cerevisiae ethanol dehydrogenase I gene(ADH I),incretin expression vector p NK1-L-PGK-10 rol GLP-1-R and Cas9 expression vector p GADT7-Cas9 were constructed and co-transformed S.cerevisiae.By using SDS-PAGE and Western Blotting analysis,the hypoglycemic S.cerevisiae(HS)with the incretin gene(10rol GLP-1)knock-in the ADH I target site was obtained,and the hypoglycemic effect of which was checked by mice model.(3)Using the synthetic Vanadyl Trehalose(VT)and DM mice models revealed that VT has "insulin-like" properties,effectively regulates glucose metabolism and lowers blood glucose,but produces certain oxidative stress damage to the gastrointestinal tract of mice by affecting GSH/GSTs pathways,Keap1-Nrf2-ARE pathway related gene expression;The reductive glutathione(GSH)combined vitamin C in formulate method was able to effectively eliminate the toxic side effects of VT,protect the gastrointestinal tract function.The results obtained above may set the foundation to development the incretin biodrugs and insulin-mimetic small molecule medicine for type 2 diabetes mellitus.