Effect Of PDX Gene 1 In Human Adipose-derived Mesenchymal Stem Cells Differentiating Into Insulin-secreting Cells

【Objective】To invesitigate the effect of PDX gene 1(PDX-1)in human adipose-derived mesenchymal stem cells(h ADSC)differentiating into insulin-secreting cells.【Methods】The adipose tissue-derived mesenchymal stem cells were randomly divided into experimental group and control group.The experimental group was treated with MOI(infected complex)= 50 in the pancreaticoduodenal homologous box gene-1,After 4 hours of culture,the culture solution containing PDX-1 gene was added and the fresh culture medium was fresh and the culture was continued for 48 hours to ensure transfection.The same method was used in the control group without adenovirus infection.Two days after infection,the results of transfection were detected by Western blotting and immunofluorescence staining.The cells were observed 15 days after staining with dithizone(DTZ)staining to count the number of cells with positive insulin expression.The secretion of insulin was detected by enzyme-linked immunosorbent assay(ELISA).The expression of insulin-1 gene was detected by immunofluorescence staining.【Results】(1)The expression of PDX-1 was detected by Western blot and immunofluorescence after adipogenic mesenchymal stem cells(PDX-1)for 48 h,and no PDX-1 expression was observed in the control group.The expression of PDX-1 was detected in the control group.(2)The insulin-positive cells were not found in the control group,and the dithizone-positive cells were stained in the experimental group.(3)After 15 days of transfection,the cells in the experimental and control groups were secreted(6.98±0.72)、(6.78±0.54)ng/m L,respectively.There was no significant difference between the two groups(P> 0.05).The amount of insulin in the high glucose environment was(13.38±0.66)、(6.50 ± 0.64)ng / m L,which was statistically significant compared with the control group,P<0.05.The expression of insulin 1 in the experimental group was higher than that in the control group.【Conclusion】The introduction of pancreaticoduodenal homologous box gene-1 promoted the differentiation of human adipose-derived mesenchymal stem cells into pancreatic islet-like cells and laid a foundation for the development of islet transplantation for type 1 diabetes mellitus.