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目的探讨在离体细胞培养条件下,Aβ1-42对SD乳鼠少突胶质前体细胞(oligodendrocyte precursor cells,OPCs)增殖与分化的影响。方法采用条件培养液原代培养获得高纯度SD乳鼠OPCs,分为正常组、加入无菌水的对照组、加入Aβ1-42的实验组,采用CCK-8检测存活细胞数量和增殖水平,Western blot检测细胞凋亡蛋白含量,免疫细胞化学染色观察细胞形态变化及分化情况。结果与对照组相比,实验组OPCs存活率显著降低(P<0.01),具体表现为:Aβ1-42作用OPCs 1 h后,细胞增殖明显受到抑制(P<0.01);药物浓度1 mol/L作用于OPCs时,增殖也受到影响(P<0.05),当Aβ1-42剂量增加至2.5 mol/L时,细胞增殖明显受到抑制(P<0.01),而对照组与正常组无显著差异(P>0.05)。对凋亡蛋白Caspase-3的检测发现,它们在对照组与正常组的表达水平基本相当(P>0.05),而在实验组明显升高(P<0.01),且呈现时间和浓度依赖性,具体表现为:Aβ1-42作用OPCs 2 h后,Caspase-3表达水平明显上调(P<0.01);药物浓度2.5 mol/L作用于OPCs时,蛋白表达水平有所升高(P<0.05),当Aβ1-42剂量增加至5 mol/L时,Caspase-3表达水平明显上调(P<0.01)。对成熟少突胶质细胞标记物髓鞘碱性蛋白的免疫染色发现,与对照组相比,实验组的细胞体积明显变小,突起明显减少。结论 Aβ1-42可能通过促使细胞凋亡而抑制OPCs增殖,并抑制了OPCs向成熟少突胶质细胞分化,从而影响中枢神经系统的髓鞘形成。
Objective To investigate the effects of Aβ1-42 on the proliferation and differentiation of oligodendrocyte precursor cells (OPCs) in SD neonatal rats in vitro. Methods Primary culture of conditioned medium was used to obtain OPCs of high purity SD suckling mice. The cells were divided into normal group, control group with sterile water, control group with Aβ1-42, CCK-8 to detect the number and proliferation of surviving cells, Western The levels of apoptosis proteins were detected by Western blotting. The morphological changes and differentiation of cells were observed by immunocytochemistry. Results Compared with the control group, the survival rate of OPCs in the experimental group was significantly decreased (P <0.01). The results showed that the proliferation of OPCs was significantly inhibited after treated with Aβ1-42 for 1 h (P <0.01) (P <0.05). When the dose of Aβ1-42 was increased to 2.5 mol / L, the cell proliferation was significantly inhibited (P <0.01), but there was no significant difference between the control group and the normal group (P > 0.05). The expression of Caspase-3 was found to be similar in the control group and the normal group (P <0.05), but significantly increased in the experimental group (P <0.01) in a time and concentration dependent manner. The specific expression of Caspase-3 was up-regulated after Aβ1-42 was treated with OPCs for 2 h (P <0.01). The protein expression was increased when treated with 2.5 mol / L OPCs (P <0.05) When the dose of Aβ1-42 was increased to 5 mol / L, Caspase-3 expression was significantly up-regulated (P <0.01). Immunostaining of myelin basic protein, a marker of mature oligodendrocyte, found that the cell volume of the experimental group was significantly smaller and the number of protuberances was significantly reduced compared with the control group. Conclusion Aβ1-42 may inhibit the proliferation of OPCs by inducing apoptosis, and inhibit the differentiation of OPCs into mature oligodendrocytes and thus affect the myelination of the central nervous system.