The Preparation Of Transgenic HIAPP Gene Mice And Phenotypic Analysis

Diabetes is a polygenic disease, and it is closely related to the environment, Thehuman islet amyloid polypeptide (human islet amyloid polypeptide, hIAPP) gene controlsthe synthesis of IAPP precursor in pancreatic β cells. It is clear that IAPP is a normalproduct of β cell secretory, while in vitro and in vivo, high-sugar high-fat and otherirritants can affect the release of IAPP. Islet amyloid can replace β cells, and islet amyloidfibers can directly damage islet cells. This study successfully prepared hIAPP transgenicmice, and made relativephenotype analysis which provides a statistic and theory basis forstudying diabetes mechanism.We successfully prepared transgenic Hiapp mice and obtained12positive F1mice,and3negative mice.We feed F1mice on the day of twelve weeks with high fat and sugardiet as well as normal diet, and do experiment such as weight test, glucose tolerance test,insulin resistance test. We kill F1mice to do experiment of quantitative PCR expression ofhIAPP, and Western blot at the68thweek.What is more, we conduct experiment of copynumber test of F1hIAPP positive mice. Afterwards, we conduct biochemical indexdetection experiments, such as triglycerides, blood glucose, non-free type fatty acids, andinsulin. Then, we do insulin, hIAPP immunohistochemistry test, pancreas HE staining.The results show that hIAPP transgenic mice fluorescence quantitative expressionlevel and copy number141,113,125,194,195,72,64,62, were significantly higher than thenegative mice,0; Immunohistochemistry results show that positive F1mice(8-6，8-8，7)accumulated lots of islet amyloid polypeptide pancreatic, and compensatory insulinsecretion occurs. Islet size becomes larger, and the number of islets as well as the functionof insulin secretion decreases; Transgenic mice(8-6，8-7，8-8) glucose tolerance decreasedsignificantly with a high blood glucose level compared to the negative mice(14，15，8-24),and insulin resistance symptoms occur. Blood biochemical indicators show that bloodglucose level in positive mice(8-6，8-7，8-8，7，110，8-20) is higher than the negativemice(8-24，15，14，8-28，44，8-27). The insulin level of positive mice(8-6，8-7，8-8，7，110，8-20) is much higher than the negative ones(8-24，15，14，8-28，44，8-27), andinsulin resistance symptoms occur. Non-free-type fatty acid and triglyceride levels aremuch higher in high fat and high sugar-induced mice(8-6，8-7，8-8，8-24，14，15) comparedwith ordinary feed(110，7，8-20，8-28，44，8-27),P<0.01suggesting that high fat and sugardiet plays a role in developing into symptoms of diabetes in mice. HE staining showedrelatively high fat and sugar diet-induced positiv（e8-6，8-7，8-8）and negative mic（e8-24，14，15). Cell disintegrate, cytoplasm dissolute, β cell outline is unclear, and the numberreduced; Fuzzy Pack nuclear structure, exocrine acinar becomes smaller, acinar cells turn smaller size, cell edema.hIAPP transgenic mice preparation succeed, and glucose level as well as insulin levelincrease, insulin resistance occurs.The study provides a statistic and theory basis forstudying diabetes mechanism and animal diabetes model.