花生四烯酰多巴胺对H2O2诱导SH-SY5Y细胞氧化应激损伤的保护作用

目的 探讨花生四烯酰多巴胺(N-arachidonoyl dopamine,NADA)对H₂O₂诱导的人神经母细胞瘤SH-SY5Y细胞氧化损伤的保护作用。方法 H₂O₂处理SH-SY5Y细胞制作细胞氧化损伤模型,通过CCK8法检测细胞活力,明确不同水平的NADA对细胞存活的影响,分析NADA对SH-SY5Y细胞的保护作用; 利用生化方法检测丙二醛(MDA)和乳酸脱氢酶(LDH)的水平; 运用DCFH-DA荧光探针检测细胞内氧自由基(ROS)水平; Hoechst33342/PI双染法检测NADA对细胞氧化损伤的保护作用; 蛋白免疫印迹实验检测抗凋亡蛋白bcl-2的表达水平。结果 H₂O₂处理SH-SY5Y细胞可导致细胞活力降低,增加MDA水平和LDH活力,促进ROS的产生,降低抗凋亡蛋白bcl-2的表达,加剧细胞凋亡。NADA可剂量依赖性提高H₂O₂诱导的SH-SY5Y细胞的存活率、降低MDA水平和LDH活力以及ROS的产生,促进抗凋亡蛋白bcl-2的表达,降低细胞凋亡。结论 NADA对H₂O₂诱导的细胞氧化损伤模型具有保护作用,其机制可能是通过抑制胞内氧化应激,促进抗凋亡蛋白bcl-2的表达,从而减少神经细胞凋亡。

Protective effect of N-arachidonoyl dopamine on H2O2 -induced injury of SH-SY5Y cells

ObjectiveTo investigate the protective effect of N-arachidonoyl dopamine(NADA)on H₂O₂-induced injury of SH-SY5Y cells.Methods SH-SY5Y cells were exposed to H₂O₂. The cell viability was determined by cell counting kit-8(CCK-8)assay. The effect of NADA on the cell viability of H₂O₂ -treated cells was determined by CCK-8 assay. The levels of malondialdehyde(MDA)and lactate dehydrogenase(LDH)were determined by biochemical method. The level of intracellular oxygen species(ROS)was determined by using a fluorescent probe-DCFH-DA. The cell apoptotic rate was determined by Hoechst33342/PI staining. Western blot was used to test the level of anti-apoptotic proteinbcl-2.Results The cell viability was significantly decreased after H₂O₂ treatment. H₂O₂ treatment resulted in higher levels of LDH and MDA in medium, and higher level of intracellular ROS. The expression level of bcl-2 was decreased after H₂O₂ treatment. H₂O₂ also induced apoptosis of SH-5Y5Y cells. NADA increased the cell viability of H₂O₂ -treated SH-SY5Y cells, decreased the levels of LDH, MDA and ROS. NADA also promoted the expression of anti-apoptotic protein bcl-2 in a dose-dependent manner. Furthermore, NADA decreased apoptotic rate of SH-5Y5Y cells.Conclusion NADA exerted a protective effect against H₂O₂ -induced SH-SY5Y cell injury through suppressing intracellular oxidative stress and promoting the expression of bcl-2.