trichostatin A对多巴胺能神经细胞的影响

发布时间：2018-06-22 05:05

本文选题：多巴胺能神经元 + TSA　；参考：《广东医学》2015年09期

【摘要】：目的探讨组蛋白去乙酰化酶抑制剂trichostatin A(TSA)对多巴胺能神经细胞的影响。方法将体外培养多巴胺能神经细胞SH-SY5Y分为6组,DMSO处理为对照组,不同浓度(50、100、200、500、1 000 nmol/L)TSA处理为TSA组,处理48 h后行MTT法检测细胞活力;采用200 nmol/L TSA处理细胞不同时间段,MTT法检测细胞活力,Western blot检测Fra1表达;构建腺病毒载体过表达Fra1或对照GFP,检测正常或TSA处理条件下过表达Fra1对细胞活力的影响。结果对照组与50 nmol/L TSA组比较,细胞存活率差异无统计学意义,与100 nmol/L TSA组比较,细胞活力明显减少(P0.05),随TSA浓度升高,活力减少更显著(P0.05);200 nmol/L TSA处理细胞8 h未检测到抑制细胞存活(P0.05),处理16 h有显著抑制作用,处理时间越久抑制效应越明显(P0.05);Western blot结果显示,TSA处理6 h未抑制Fra1表达(P0.05),处理12 h明显抑制Fra1表达,24 h抑制效应更显著(P0.05);过表达Fra1促进细胞存活(P0.05),显著拮抗TSA的抑制存活效应(P0.05)。结论 TSA抑制多巴胺能神经细胞存活通过抑制Fra1表达。
[Abstract]:Objective to investigate the effect of histone deacetylase inhibitor trichostatin A (TSA on dopaminergic neurons. Methods the dopaminergic neurons (SH-SY5Y) were divided into 6 groups (control group) treated with DMSO and treated with TSA at different concentrations (50,100g / L, 500,000nmol / L). After 48 h treatment, the cell viability was measured by MTT assay. The expression of Fra1 was detected by using 200nmol / L TSA at different times, and the effect of overexpression of Fra1 on cell viability under normal or TSA-treated condition was detected by constructing adenovirus vector overexpression Fra1 or control GFP. Results there was no significant difference in cell survival rate between the control group and the 50 nmol / L TSA group. Compared with the 100 nmol / L TSA group, the cell viability was significantly decreased (P0.05), and the cell viability increased with the increase of TSA concentration. The activity decrease was more significant (P0.05) after treatment with 200 nmol / L TSA for 8 h, the inhibition of cell survival was not detected (P0.05), but it was significantly inhibited at 16 h after treatment. The results of Western blot showed that TSA did not inhibit the expression of Fra1 for 6 h (P0.05), significantly inhibited the expression of Fra1 for 24 h (P0.05), over-expressed Fra1 promoted cell survival (P0.05), and significantly antagonized the inhibitory effect of TSA (P0.05). Conclusion TSA inhibits the survival of dopaminergic neurons by inhibiting Fra1 expression.
【作者单位】：广州医科大学附属第二医院神经外科;广州医科大学神经科学研究所(广东省重点实验室神经遗传与离子通道病省部共建教育部重点实验室);
【基金】：国家自然科学基金资助项目(编号:31171021) 广东省中医药局建设中医药强省科研课题(编号:20121135) 广东省自然科学基金资助项目(编号:S2011010003392) 广州市属高校科研项目(编号:10A223)
【分类号】：R96

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