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目的探讨白藜芦醇预处理对大鼠皮质神经元氧糖剥夺/再复氧损伤后神经元突起生长的影响。方法体外皮质神经元氧糖剥夺150min,复氧培养24 h。实验分为正常组、对照组和5μmol/L白藜芦醇预处理组。免疫荧光法鉴定神经元,细胞计数试剂盒8(CCK-8)法测定细胞活力,TUNEL法检测细胞凋亡,免疫荧光法和Western blotting检测微管相关蛋白2(MAP-2)和生长相关蛋白43(GAP-43)的表达,并计数神经元突起的长度和数目。结果培养细胞均高表达神经元特异性标记物MAP-2。白藜芦醇组细胞活力较对照组明显增加(0.551±0.009比0.436±0.013,P<0.01),而凋亡则明显降低(18.3%±1.3%比35.3%±1.9%,P<0.01),上调MAP-2(0.790±0.102比0.462±0.063,P<0.01)和GAP-43(0.768±0.084比0.424±0.065,P<0.01)蛋白的表达,增加神经元突起的长度(89.510±6.939比61.538±9.14,P<0.01)和数目(6.347±1.002比3.040±0.608,P<0.01)。结论白藜芦醇预处理能减轻氧糖剥夺/再复氧对神经元的损伤,促进神经元突起的生长。
Objective To investigate the effect of resveratrol preconditioning on the neurite outgrowth after oxygen glucose deprivation / reoxygenation injury in rat cortical neurons. Methods Cortical neurons were deprived of oxygen for 150 minutes and reoxygenated for 24 hours. The experiment was divided into normal group, control group and 5μmol / L resveratrol pretreatment group. Neurons were identified by immunofluorescence, cell viability was determined by CCK-8 assay, apoptosis was detected by TUNEL, and MAP-2 and growth-related proteins 43 (GAP-43) and counted the length and number of neuronal processes. Results The cultured cells all highly expressed neuron-specific marker MAP-2. The viability of resveratrol group was significantly higher than that of control group (0.551 ± 0.009 vs. 0.436 ± 0.013, P <0.01), while the apoptosis was significantly decreased (18.3% ± 1.3% vs 35.3% ± 1.9%, P <0.01) (0.790 ± 0.102 vs 0.462 ± 0.063, P <0.01) and GAP-43 (0.768 ± 0.084 vs 0.424 ± 0.065, P <0.01) increased the neurite outgrowth length (89.510 ± 6.939 vs. 61.538 ± 9.14, P <0.01) and the number (6.347 ± 1.002 vs. 3.040 ± 0.608, P <0.01). Conclusion Resveratrol preconditioning can reduce the damage of neurons induced by oxygen deprivation / reoxygenation and promote neurite outgrowth.