摘要
目的:探讨丹参注射液在体外诱导大鼠骨髓间充质干细胞(rM SC s)向神经元样细胞分化的作用。方法:采用贴壁筛选法分离rM SC s,并通过不断传代进行纯化和扩增培养。采用含丹参注射液的无血清Du lbecco改良E ag le培养基(DM EM)/F 12培养液诱导rM SC s分化为神经元样细胞,观察不同浓度丹参注射液对rM SC s诱导分化为神经元样细胞的影响。通过观察rM SC s经诱导后细胞的形态变化和采用免疫组化方法检测神经元特异性核蛋白(N euN)、神经丝蛋白(NF M)、胶质纤维酸性蛋白(GFAP)的表达来鉴定神经元样细胞。结果:经丹参注射液诱导后,rM SC s胞体收缩,突起伸出,形似神经元;免疫组化显示诱导出的神经元样细胞N euN、NF M表达阳性,GFAP阴性。丹参注射液在浓度为10 m l/L时的诱导作用最为显著。结论:丹参注射液可以在体外诱导rM SC s分化为神经元样细胞,其诱导分化率与丹参注射液的浓度有关。
Objective To investigate the effects of differentiation of rat bone mesenchymal stem cells （rMSCs） into neuron-like cells with salvia miltiorrhiza injection in vitro. Methods rMSCs were separated from rat bone marrow with wall sticking method and expanding, and cultured in vitro, rMSCs were induced to differentiate into neuron-like cells with salvia miltiorrhiza injection. The effects of different dosages of salvia miltiorrhiza on differentiation of rMSCs into neuron-like cells were observed. Neuron-like cells were identified with neuron-specific enolase （NeuN）, neurofilament （NF- M）, glial fibrillary acidic protein （GFAP） by immunohistochemistry. Results The rMSCs were induced by salvia miltiorrhiza injection to express neuronal phenotype similar to the cell appearance of neurons with NeuN and NF - M positive at 3 hours, but glial astrocyte marker GFAP negative. The differentiation was the most significant when the inducing concentration of salvia miltiorrhiza injection was at 10 ml/L. Conclusion rMSCs can be induced to differentiate into neuron-like cells with salvia miltiorrhiza injection. The efficacy of differentiation is correlated with the concentration of salvia miltiorrhiza injection.
出处
《中国中西医结合急救杂志》
CAS
2006年第4期210-213,F0002,共5页
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
基金
国家中医药管理局基金资助项目(2003LHR11)
浙江省科技厅基金资助项目(2004C33014)
关键词
骨髓间充质干细胞
神经元
丹参注射液
诱导分化
大鼠
bone mesenchymal stem cells
neuron
salvia miltiorrhiza injection
induction and differentiation
rat
