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目的探讨高良姜素对多巴胺诱导的A375黑素细胞凋亡及其Bcl-2基因转录水平的影响。方法利用1 mmol/L的多巴胺处理A375细胞24 h后,以0.5,1.0,2.0μg/ml的高良姜素作用于受损的A375细胞。采用MTT法检测细胞存活率变化,TBA法试剂盒检测细胞中MDA含量,WST法测定细胞SOD酶活性。高良姜素作用A375细胞48h后,收集细胞,分别提取其基因组DNA和总RNA,DNA Ladder法检测药物对受损A375细胞凋亡的影响,通过半定量RT-PCR测定给药后各组的光密度值,计算各组的Bcl-2/GAPDH值,从而分析Bcl-2基因转录水平的变化。结果高良姜素各剂量组均可显著提高受损A375细胞存活率,降低细胞中MDA生成及促进SOD酶活性。0.5μg/ml的高良姜素对A375细胞凋亡保护不明显,随着给药浓度的增大,其细胞凋亡作用逐渐增强。此外,0.5,1.0,2.0μg/ml的高良姜素对A375细胞Bcl-2基因的转录均具有上调作用。结论高良姜素对多巴胺诱导的A375细胞凋亡具有显著保护作用,其作用机制可能与其降低细胞MDA产生、促进SOD酶活性及上调Bcl-2基因的转录水平有关。
Objective To investigate the effect of galangin on apoptosis of dopamine induced A375 melanocytes and the expression of Bcl-2 gene. Methods A375 cells were treated with 1 mmol / L dopamine for 24 h, then treated with galangin at concentrations of 0.5, 1.0 and 2.0 μg / ml for A375 cells. Cell viability was detected by MTT assay, MDA content was detected by TBA method and SOD activity by WST method. After galangin A375 cells were treated for 48h, the cells were harvested and their genomic DNA and total RNA were extracted respectively. The effects of drugs on the apoptosis of A375 cells were detected by DNA Ladder method. The light of each group was determined by semi-quantitative RT-PCR Density, calculate the Bcl-2 / GAPDH value of each group to analyze the change of Bcl-2 gene transcription level. Results All doses of galangin significantly increased the survival rate of damaged A375 cells, decreased the production of MDA and promoted the activity of SOD. 0.5ng / ml of galangin had no significant protective effect on A375 cells apoptosis, with the increase of the concentration, the apoptosis of A375 cells gradually increased. In addition, galangin at 0.5, 1.0 and 2.0 μg / ml up-regulated the transcription of Bcl-2 gene in A375 cells. CONCLUSION Gallicin has a significant protective effect on the apoptosis of A375 cells induced by dopamine. Its mechanism may be related to the decrease of cell MDA, the activity of superoxide dismutase and the up-regulation of Bcl-2 gene transcription.