Trichostatin A promotes mouse bone marrow mesenchymal stem cells to differentiate into insulin-secreting cells / 中国病理生理杂志

ABSTRACT

AIM:To investigate whether trichostatin A ( TSA) , a new revulsant ,can induce mouse mesenchy-mal stem cells to differentiate into insulin-secreting cells and to explore the appropriate concentration of TSA .METHODS:The mesenchymal stem cell line from C57BL/6 mice was cultured in vitro and divided into 5 groups before treated with dif-ferent concentrations of TSA , ( group A:DMSO;group B~E:treated with 25 nmol/L, 50 nmol/L, 100 nmol/L and 200 nmol/L of TSA, respectively).After exposed to different cultured media for 10 d during the 2 stages, the cells were detec-ted by the following methods:the insulin-secreting cells in each group were identified by dithizone staining and the results were calculated with immunohistochemical half quantitative analysis .The insulin secreted by insulin-secreting cells in each group was identified by immunofluorescence , and the mean fluorescence intensity of insulin was compared .The content of insulin in each group was quantified by ELISA .The appropriate concentration of TSA was determined according to the above results .RESULTS:TSA treatment for 10 d promoted the mouse bone marrow mesenchymal stem cells to differenti-ate into insulin-secreting cells which produced insulin .The immunohistochemistry and immunofluorescence imaging analysis of insulin-secreting cells showed that the insulin staining positive area , positive ratio , total density of insulin expression and mean fluorescence intensity of insulin in group B were significantly higher than those in the other TSA -treated groups .When the concentrations of TSA gradually increased , the content of insulin reduced accordingly .The content of insulin in group B was significantly higher than that in the other TSA-treated groups .CONCLUSION:TSA treatment for 10 d promotes bone marrow mesenchymal stem cells from C57BL/6 mice to differentiate into insulin-secreting cells and the appropriate concen-tration of TSA is 25 nmol/L.