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目的 用川芎嗪 (ligustrazinhydrochloride)在体外定向诱导SD青年鼠骨髓间质干细胞 (mesenchymalstemcells ,rMSCs)分化为神经元样细胞。 方法 用低糖DMEM冲洗骨髓腔 ,收集骨髓细胞悬液 ,接种在塑料培养瓶中。经体外扩增、纯化 ,选用第 5代后的骨髓间质干细胞进行诱导分化。用 10 μg LbFGF预诱导 2 4h ,更换成含川芎嗪的无血清培养基DMEM诱导间质干细胞分化为神经元样细胞。用免疫组织化学SABC法鉴定神经丝蛋白 (NF M)、神经元特异性烯醇化酶 (NSE)、巢蛋白 (nestin)、微管联合蛋白 2 (MAP 2 )、生长相关蛋白 4 3(GAP 4 3)、胶质纤维酸性蛋白 (GFAP)的表达。 结果 第 5代间质干细胞形态达到均一 ,呈梭形。用川芎嗪诱导 15min到 3h ,间质干细胞胞体逐渐增大 ,并伸出细长突起形似神经元样细胞。免疫组织化学显示NF M、NSE、nestin、MAP 2和GAP 4 3表达阳性 ,而GFAP阴性。对照组上述染色均为阴性。 结论 川芎嗪可诱导骨髓间质干细胞分化为神经元样细胞。
Objective To induce differentiation of mesenchymal stem cells (rMSCs) in SD young rats into neuron-like cells in vitro induced by ligustrazine hydrochloride. Methods Bone marrow cavity was washed with low-glucose DMEM, bone marrow cell suspension was collected and inoculated in a plastic culture flask. After in vitro expansion and purification, the fifth generation of bone marrow mesenchymal stem cells were used to induce differentiation. 10 μg LbFGF was pre-induced for 24 hours and replaced with a serum-free medium containing Ligustrazine to induce differentiation of mesenchymal stem cells into neuron-like cells. Identification of neurofilament protein (NF M), neuron-specific enolase (NSE), nestin, microtubule associated protein 2 (MAP 2), growth-associated protein 4 3 (GAP 4) by immunohistochemical SABC method 3) Expression of glial fibrillary acidic protein (GFAP). Results The morphology of the passage 5 mesenchymal stem cells was uniform and fusiform. Induced by tetramethylpyrazine for 15 minutes to 3 hours, the mesenchymal stem cell body gradually increased, and elongated elongated protrusions resembled neuron-like cells. Immunohistochemistry showed positive expression of NF M, NSE, nestin, MAP 2 and GAP 4 3, whereas GFAP was negative. In the control group, the above staining was negative. Conclusion Tetramethylpyrazine can induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.