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背景:尿酸作为一种内源性的抗氧化剂,其抗氧化、抗DNA损害作用及发挥神经元保护作用近年受到关注。目的:观察不同浓度尿酸对骨髓间充质干细胞成神经分化的影响。方法:体外分离、纯化、培养骨髓间充质干细胞,观察细胞形态,取扩增3代的骨髓间充质干细胞,分别用含0(对照组)、0.2,0.4,0.8 mmol/L浓度尿酸的预诱导液诱导24 h,再用含0(对照组)、0.2,0.4,0.8 mmol/L浓度尿酸的诱导液诱导1 h后行尼氏体染色,2 h后,用免疫组织化学法检测细胞内巢蛋白、神经元特异性烯醇化酶的表达。结果与结论:骨髓间充质干细胞经诱导后,细胞胞体收缩,形成突起并形成连接。细胞胞质中存在蓝色颗粒状的尼氏体,含不同浓度尿酸的诱导组神经元特异性烯醇化酶阳性率均较对照组明显升高(P<0.05),而且随着尿酸浓度增加,神经元特异性烯醇化酶阳性表达率逐渐增加(P<0.05)。含不同浓度尿酸的诱导组巢蛋白阳性表达率较对照组降低(P<0.05),诱导4 h后,细胞脱落明显。在体外一定时间内,尿酸能够促进骨髓间充质干细胞向神经元样细胞的分化,且具有一定的浓度依赖性。
BACKGROUND: Uric acid, as an endogenous antioxidant, has received much attention in recent years for its antioxidant, anti-DNA damage and neuroprotective effects. Objective: To observe the effects of different concentrations of uric acid on the neural differentiation of mesenchymal stem cells. Methods: Bone marrow mesenchymal stem cells (MSCs) were isolated, purified and cultured in vitro. The morphological changes of MSCs were observed. The bone marrow mesenchymal stem cells (MSCs) of three generations were amplified and the cells were treated with 0 (control), 0.2, 0.4 and 0.8 mmol / The pre-induced solution was induced for 24 h, then induced by uracil containing 0 (control group), 0.2, 0.4 and 0.8 mmol / L uric acid for 1 h, then Nissl staining was performed. After 2 h, the cells were detected by immunohistochemistry Nestin, neuron-specific enolase expression. RESULTS AND CONCLUSION: After induction of bone marrow mesenchymal stem cells, the soma of cells contracted to form protuberances and form connections. The presence of blue granular Nissl bodies in the cytoplasm of the cells showed that the positive rate of neuron-specific enolase in the induction group with different concentrations of uric acid was significantly higher than that in the control group (P <0.05). With the increase of uric acid concentration, The positive rate of neuron specific enolase increased gradually (P <0.05). The expression of nestin in the induced group with different concentrations of uric acid was lower than that in the control group (P <0.05). After 4 h of induction, the cells shed off obviously. In a certain period of time in vitro, uric acid can promote the differentiation of BMSCs into neuron-like cells in a concentration-dependent manner.