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【目的】观察低氧对体外培养的人骨髓间充质干细胞(hBMSCs)向胆碱能神经元分化的影响。【方法】将对照组和常氧分化组hBMSCs置于常氧(20%O2)环境中培养,低氧分化组细胞置于低氧(3%O2)环境中。采用免疫细胞化学法和高效液相色谱—电化学法,分别检测胆碱能神经元的数量和诱导分化后乙酰胆碱(ACh)的含量。【结果】对照组未见胆碱乙酰化酶(ChAT)阳性神经元,低氧分化组ChAT阳性神经元数量明显增多,约为常氧分化组的4倍(P<0.01),并且该组诱导分化后的细胞合成的ACh含量也高于常氧分化组(P<0.05)。【结论】低氧可促进hBMSCs向胆碱能神经元方向分化,这为临床应用hBMSCs治疗阿尔茨海默病提供了新的思路。
【Objective】 To observe the effect of hypoxia on cholinergic neuronal differentiation of hBMSCs cultured in vitro. 【Methods】 The control and normoxia groups were cultured in normoxia (20% O2) environment, hypoxia cells were exposed to hypoxia (3% O2). The number of cholinergic neurons and the content of acetylcholine (ACh) after differentiation were detected by immunocytochemistry and high performance liquid chromatography-electrochemistry respectively. 【Results】 ChAT-positive neurons were not found in the control group. The number of ChAT-positive neurons in the hypoxia group was significantly increased, which was about 4-fold (P <0.01), compared with that in the normoxia group ACh content in differentiated cells was also higher than that in normoxia group (P <0.05). 【Conclusion】 Hypoxia can promote hBMSCs to differentiate into cholinergic neurons, which provides a new idea for the clinical application of hBMSCs in the treatment of Alzheimer’s disease.