【摘要】 目的:研究复方丹参注射液体外诱导人胚胎和成年小鼠骨髓基质干细胞(MSCs)体外定向神经细胞发育的规律。方法:应用复方丹参注射液诱导体外培养的人胚胎和成年小鼠骨髓基质干细胞,观察诱导过程中的形态学变化,并于诱导后5、24、72 h分别应用抗Nestin、抗bⅢ-tubulin、抗MBP及抗GFAP抗体进行免疫细胞化学染色,RT-PCR检测mRNA表达,Westem Blot检测蛋白表达。应用5mmol/L和10 mmol/L的2-巯基乙醇作为诱导对照组。结果:复方丹参注射液和2-巯基乙醇均可诱导骨髓基质干细胞分化为神经样细胞,但复方丹参注射液诱导作用更为缓和,细胞死亡率远低于2-巯基乙醇诱导组。免疫细胞化学示Nestin和bⅢ-tubulin呈阳性表达,MBP及GFAP呈阴性表达。RT-PCR可见Nestin和bⅢ-tubulin mRNA呈阳性表达,Westem Blot检测证实,Nestin和bⅢ-tubulin蛋白呈阳性表达。结论:复方丹参注射液可在体外诱导人胚胎和成年小鼠骨髓基质干细胞向神经细胞发育,是较为理想的诱导剂。更多还原

【Abstract】 Objective: To investigate the regulation of neuronal development of marrow stromal stem cells(MSCs) induced by salvia miltiorrhiza and b-mercaptoethanol in human fetal and Kunming adult mouse in vitro.Methods: Morphological changes of neuronal development during the induction of MSCs established by salvia miltiorrhiza were observed and compared with those induced by 5 mmol/L,10 mmol/L b-mercaptoethanol as control.Cells were stained immuneocytochemically with Nestin,bⅢ-tubulin,MBP and GFAP 5 hours,24 hours and 72 hours after induction,the mRNA was determined by RT-PCR and the expression of protein was detected by Westem blotting.Results: Typical neuron-like morphology appeared gradually with the extension of induction,whereas mortality rate of MSCs caused by b-mercaptoethanol was really higher than that by salvia miltiorrhiza.Cells were stained positively with Nestin and bⅢ-tubulin antibody but negatively with MBP and GFAP.Nestin and bⅢ-tubulin mRNA and protein positively expressed by RT-PCR and Western blotting.Conclusion: Marrow stromal stem cells in human fetal and adult mouse can develop into neural cells by salvia miltiorrhiza and b-mercaptoethanol in vitro.Salvia miltiorrhiza is superior to b-mercaptoethanol.更多还原