Transplantation Effects Of IPMSCs Regulated By Conophylline Via PI3K Pathway In Diabetic Mice

Study found that small molecular substance named conophylline, an alkaloid isolated from the leaves of Ervatamia microphylla, which can promote pancreatic stem cells proliferation differentiation. In this study, the immortalized mesenchymal stem cells derived from fetal porcine pancreas, iPMSCs were utilized as the experimental subject to investigate the function of conophylline. Simultaneously to explore the implementation of proliferation and differentiation was carried out whether though PI3K signal pathway, and then transplantation induced iPMSCs into diabetic mice, suggested that the conophylline could promote differentiation, simultaneously, provide a theoretical principle for small molecules promoting pancreatic cell differentiation.1, Conophylline promote iPMSCs proliferation via PI3K.In order to demonstrate the influence of conophylline for iPMSCs proliferation, conophylline and LY-294002(the inhibitor of PI3K) were added in the culture medium of iPMSCs respectively, and then the growth curve and state had been observed, expression of genes such as PCNA, C-myc, PDX1, Glut2and Insulin could also be detected. The results suggested that conophylline could promote proliferation of iPMSCs, increase the efficiency of DNA synthesis, and the cells of S phase were significant more than other groups, the most important, that the IF and WB consist with the previous results. On the contrary, LY could inhibit the function of conophylline. The most important is that expression of Glut2and Insulin could not be detected in all groups.2, Conophylline promoted iPMSCs differentiate to insulin-secretory cells in vitro.We assembled2step induced cells which were added conophylline and LY to medium of iPMSCs respectively. And then target genes such as PDX1,Glut2and Insulin had been detected by RT-PCR and WB; observed the synthesis of insulin for sensitive of high-sugar. Consequently, solo conophylline could not significantly induce iPMSCs differentiate to islet cell clusters; up-regulate the expression level of PDX1、Glut2and Insulin. Positive DTZ reaction suggests that the induced iPMSCs are sensitive for high sugar:insulin secretion, C-peptide formation; on the contrary, the function of LY group was significantly reduced.3, The induced iPMSCs transplant into diabetic mice.Firstly, there is no tumorigeniciry proved by induced iPMSCs transplantation in nude mouse by1×107, therefor it is secure for clinical application. Induced iPMSCs transplanted into diabetic mice, the results revealed that reverse hyperglycaemia could be temporarily relieved by transplantation of induced iPMSCs.It is demonstrated that conophylline plays a key role in the regulation of cell proliferation, promotion of expression of pancreatic function related genes. Induced iPMSCs could secrete insulin and response to high glucose level, and transplanted iPMSCs could improve hyperglycaemia of diabetic mice. Thus, conophylline has the potential as novel drugs for diabetes treatment. iPMSCs from swine might be a new horizon of substitutive therapy by target cells transplantation.