Bcl-xl基因对硝普钠诱导SY5Y细胞凋亡的抑制作用  

Inhibition Effect of Bcl-xl Gene on Apoptosis of SY5Y Cells Induced by Sodium Nitroprusside

作　　者：杨炼红[1] 蒋龙元[2] 蔡晓冬[3] 刘淑琼[1] 吴秀勤[1] 刘军[1] 陶恩祥[1] 

机构地区：[1]中山大学孙逸仙纪念医院神经科,广东广州510120 [2]中山大学孙逸仙纪念医院急诊科,广东广州510120 [3]中山大学附属第一医院神经科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2012年第4期449-453,459,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省科技计划项目(2006B36004016);广东省自然科学基金(S2011010004626)

摘　　要：【目的】研究Bcl-xl基因对成人神经母细胞廇SH-SY5Y细胞毒性的影响。【方法】将构建好的Bcl-xl基因重组真核表达载体pIRES2-EGFP/Bcl-xl转染SH-SY5Y细胞,并分为正常组、pIRES2-EGFP空质粒转染组、pIRES2-EGFP/Bcl-xl质粒转染组,利用凋亡诱导剂硝普钠(50μmol/L)诱导各组细胞凋亡,行Hoechst33258细胞染色对比观察各组细胞凋亡情况,通过MTT法分析各组细胞存活率。RT-PCR与Western blotting检测转染后Bcl-xl基因的表达情况。【结果】转染SH-SY5Y细胞后,检测结果显示Bcl-xl基因能够高效转染并在此细胞中蛋白表达。相对于未转染细胞来说,转染细胞能部分对抗硝普钠诱导的凋亡,MTT显示有统计学意义(P<0.05)。【结论】Bcl-xl基因能够通过升高基因的表达抑制NO诱导的类神经元细胞死亡,同时也能间接说明硝普钠产生的NO对细胞的毒性可能大部分是通过凋亡途径来产生的。[ Objective] To study the effect of Bcl-xl gene on cytotoxicity of SH-SY5Y cells induced by sodium nitroprusside. [ Methods ] The plasmid pIRES2-EGFP/Bcl-xl was transfected into SH-SY5Y cells. Three groups were designed： A） normal control group ; B） pIRES2-EGFP control group ; C） pIRES2-EGFP/Bcl-xl group. After SH-SY5Y cells being treated with sodium nitroprusside （50 ~mol/L）, the apoptosis was valued by Hoechst33258 stain. In addition, M33＂ was employed to test cell viability. We also did RT- PCR and Western blotting to investigate Bcl-xl gene expression. [ Results ] By transfection of pIRES2-EGFP/Bcl-xl, Bcl-xl gene could effectively express in SH-SY5Y cells. Supported by MTF results, Bcl-xl gene was able to partially reduce cell death induced by sodium nitroprusside. [Conclusion] Transfection of Bcl-xl gene can increase the expression of Bcl-xl and thus lead to weaken cell death induced by NO supplied by sodium nitropmsside. This result supports that cytotoxicity induced by SNP is mainly mediated via the process of apoptosis.

关 键 词：Bcl—xl基因 SH—SY5Y 硝普钠 凋亡 基因治疗 

分 类 号：R741.02[医药卫生—神经病学与精神病学]