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目的:探讨槲皮素(Quercetin,Qu)对大鼠缺氧损伤后少突胶质前体细胞(oligodendrocyte precursor cells,OPCs)增殖的作用。方法:取新生1～2 d Sprague-Dawley(SD)大鼠大脑皮质细胞进行原代培养,振荡分离后纯化3 d行Olig2和A2B5免疫荧光染色鉴定。纯化的细胞分为正常对照组、模型组(缺氧9 h)和Qu处理组(3,9,27,81μmol/L)。缺氧后3 d光镜下观察细胞形态学变化,CCK-8法检测细胞存活率,Olig2免疫荧光染色计数阳性细胞数。结果:纯化培养3 d的细胞胞体呈圆形,有双极或三极突起。免疫荧光染色显示,绝大部分细胞为A2B5和Olig2阳性,A2B5/DAPI阳性细胞率为98.54%。缺氧9 h后,模型组很多细胞出现突起皱缩或崩解,细胞密度较Qu处理组明显降低;培养至3 d,光镜下计数和CCK-8检测结果均表明9μmol/L和27μmol/L Qu处理组细胞数较模型组显著增加(P<0.01,P<0.05);Olig2免疫细胞化学染色结果进一步表明9μmol/L和27μmol/L Qu处理组OPCs数较模型组显著增加(P<0.01,P<0.05)。结论:9μmol/L和27μmol/L Qu有促进缺氧损伤OPCs增殖的作用。
Objective: To investigate the effect of quercetin on the proliferation of oligodendrocyte precursor cells (OPCs) after hypoxia in rats. Methods: Primary cultured rat cerebral cortical neurons were isolated from Sprague-Dawley (SD) rats for 1 ~ 2 days. The cells were isolated by shaking and purified by Olig2 and A2B5 immunofluorescence staining for 3 days. The purified cells were divided into normal control group, model group (9 h hypoxia) and Qu treatment group (3, 9, 27, 81 μmol / L). Morphological changes were observed under light microscope 3 days after hypoxia. Cell viability was detected by CCK-8 assay and Olig2 immunofluorescence staining was used to count the number of positive cells. Results: The cell bodies of the cells cultured for 3 days were round and had bipolar or tripolar processes. Immunofluorescence staining showed that the majority of cells were positive for A2B5 and Olig2, and the positive rate of A2B5 / DAPI was 98.54%. After 9 hours of hypoxia, many cells in the model group showed shrinkage or disintegration, and the cell density was significantly lower than that in Qu treatment. Counting by light microscopy and CCK-8 results showed that 9μmol / L and 27μmol / Compared with model group, the number of cells in L Qu treatment group was significantly increased (P <0.01, P <0.05). The results of Olig2 immunocytochemistry further showed that the number of OPCs in 9μmol / L and 27μmol / L Qu treatment groups was significantly increased , P <0.05). Conclusion: 9μmol / L and 27μmol / L Qu can promote the proliferation of OPCs induced by hypoxia.