摘要
目的:观察曲古抑菌素A(TSA)诱导小鼠骨髓间充质干细胞分化为胰岛素分泌细胞的适宜浓度。方法:C57BL/6小鼠骨髓间充质干细胞系体外培养。实验分为5组:对照组(DMSO,A组)和TSA干预组(25 nmol/L,B组;50 nmol/L,C组;100 nmol/L,D组;200 nmol/L,E组)。各组分两阶段用相应的培养基培养10 d后进行检测。双硫腙染色鉴定各组的胰岛素分泌细胞,结果进行半定量分析。免疫荧光染色鉴定各组的胰岛素分泌,比较各组胰岛素平均荧光强度。酶联免疫吸附试验检测各组胰岛素分泌细胞胰岛素的分泌量。结果:TSA作用10 d能够诱导C57BL/6小鼠骨髓间充质干细胞分化成为胰岛素分泌细胞并分泌胰岛素。B组胰岛素染色阳性面积、阳性率、胰岛素染色积分吸光度以及胰岛素平均荧光强度显著高于其它干预组,差异有统计学意义(均P<0.05)。随着各干预组TSA浓度的升高,胰岛素的分泌量减少。B组胰岛素含量显著高于其它干预组,差异有统计学意义(均P<0.05)。结论:TSA处理10 d能诱导C57BL/6小鼠骨髓间充质干细胞分化成为胰岛素分泌细胞,并且25 nmol/L为TSA的适宜浓度。
AIM：To investigate whether trichostatin A （ TSA） , a new revulsant ,can induce mouse mesenchymal 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 different 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 detected 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 differentiate 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 TSAtreated 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 concentration of TSA is 25 nmol/L.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第6期1088-1092,共5页
Chinese Journal of Pathophysiology
