高良姜素对多巴胺诱导黑素细胞凋亡的保护作用研究

Study on protective effects of Galangin on apoptosis of melanocyte induced by Dopamine

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 and Bcl - 2 gene transcription level of A375 cell induced by Dopamine. Methods After lmMol/L Dopamine induced 24h,and then antagonize the effect of Dopamine by different concentra- tion（0.5,1.0,2.0 μg/ml）Galangin. We determinated the cell survival with MTT, measured the MDA contents and SOD activity by TBA kit and WST kit. After treated 48h with Galangin, ceils was collected, extract genome DNA and total RNA of A375 cell, through the DNA Ladder assay the apoptosis, measured light density of every group by semi - quantitative RT - PCR, and analysis the change of the transcription level of Bcl - 2 gene by calculating Bcl - 2/GAPDH of every group. Results Galangin of all dose groups could significantly improve the cell survival, inhibit the production of MDA and increase the activety of SOD. The concentration such as O. 5 μg/ml has no significantly protective effects on apoptosis, but the protective effects of other dose groups became stronger and stronger with the concentration increased. Moreover, both 0.5,1.0 and 2.0μg/ml of Galangin have up - regulated effect on the transcription level of Bcl - 2 gene. Conclusion Galangin has significantly protective effects on apoptosis of A375 cell induced by dopamine, and it′s protective mechanism probably relate to inhibit the production of MDA, increase the activety of SOD and up - regulated the transcription level of Bcl - 2 gene.