中药诱导间充质干细胞向神经细胞分化后的免疫调控作用

Neurons drived from mesenchymal stem cells with the induction of Chinese traditional herbal drugs harbor immunoregulatory effects

目的:探讨多种中药成分体外定向诱导骨髓间充质干细胞(MSCs)向神经元样细胞分化,并研究分化细胞的免疫调控能力。方法:通过贴壁法分离人骨髓间充质干细胞(hMSCs),体外扩增培养。多种中药成分定向诱导分化为神经元样细胞。光镜下观察细胞形态,免疫细胞化学检测神经细胞特异性抗原标志物神经元烯醇化酶(NSE)、巢蛋白(Nestin)、胶质纤维酸性蛋白(GFAP)的表达。通过单向混合淋巴细胞反应(MLR),观察神经细胞分化的hMSCs对人外周血T淋巴细胞(human periphera lblood lymphocyte,hPBL)免疫调控作用。结果:hMSCs可通过贴壁法成功分离并可在体外大量扩增。黄芪,天麻,人参诱导1～3小时后大部分hMSCs转变为神经元样细胞,出现胞体和突起。免疫细胞化学染色NSE及Nestin呈阳性,GFAP阴性。混合培养结果显示分化hMSCs抑制hPBLs增殖反应。结论:多种中药成分和中药制剂可在体外诱导hMSCs分化为神经元样细胞,分化hMSCs抑制人淋巴细胞增殖反应,具有免疫调控作用。

Objective：To study the immunoregulatory effect of neurons drived from mesenchymal stem cells with the induction of some Chinese traditional herbal drugs.Methods：Human mesenchymal stem cells（hMSCs） were isolated from bone marrow by wall sticking method,amplifying cultured in vitro,and differentiated into neuron-like cells by oriented induction with some Chinese traditional herbal drugs.The morphology of cells was observed under lightmicroscopy,neuron-specific enolase（NSE）,nestin and glial fibrillary acidic protein（GFAP） were detected by imnunocytochemistry.The immunoregulatory effect on T lymphocytes of the neuron-like cells induced from hMSCs were detected by one-way mixed lymphocyte reaction（MLR）.Results：hMSCs could be separated and amplified in vitro.After being induced by Astragale,Gastrodia,Ginsenosides for 1-3 h,most of the cells would be differentiated into meuron-like cells,revealing cytodendrite.By immunochemical staining,cells showed positive of NSE,nestin,and negative of GFAP.MLR assays demonstrated that neurons drived from hMSCs could suppress the proliferation of allogeic T lymphocytes.Conclusion：hMSCs can be induced to differentiate themselves into neuron-like cells by some Chinese traditional herbal drugs.Neurons drived from hMSCs could suppress the proliferation of allogeic T lymphocytes and harbor immunoregulatory effects.