PARP-1沉默对骨髓间充质干细胞凋亡的影响

发布时间：2018-01-03 22:37

本文关键词：PARP-1沉默对骨髓间充质干细胞凋亡的影响　出处：《中国老年学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的探讨沉默PARP-1对1,4-苯醌(PBQ)致大鼠骨髓间充质干细胞(BMSCs)凋亡的影响。方法以空载体BMSCs为对照组,以PARP-1沉默BMSCs为处理组,免疫印迹法检测聚腺苷二磷酸核糖聚合酶(PARP)-1蛋白表达量。磷酸盐缓冲液(PBS)溶解PBQ,分别以0、2.5、5.0、10.0、20.0、40.0、80.0、160.0和320.0μmol/L PBQ染毒两组细胞,根据细胞活力检测结果选择合适染毒浓度后,应用噻唑蓝(MTT)比色法检测细胞活力,Annexin V-PE/7-AAD标记的流式细胞技术检测细胞凋亡,实时荧光定量-聚合酶链反应检测两组细胞PARP-1基因表达。结果沉默细胞PARP-1蛋白表达量〔(0.15±0.02)倍〕与对照组细胞〔(1.00±0.03)倍〕相比,下降了85%(P0.05)。PBQ染毒24 h后,相同剂量下两组细胞的活力并无显著性差异。随着PBQ染毒剂量增加,两组细胞PARP-1表达水平和凋亡率均明显增高(P0.05),且相同剂量下PARP-1沉默细胞比对照组细胞凋亡率明显增高(P0.05)。结论 PARP-1沉默BMSCs在PBQ作用下更易发生凋亡,PARP-1可能参与了PBQ诱导的细胞凋亡。
[Abstract]:Objective to investigate the silencing of PARP-1 1,4- benzoquinone (PBQ) induced rat bone marrow mesenchymal stem cells (BMSCs) apoptosis. Methods BMSCs empty vector control group, PARP-1 silencing BMSCs as treatment group and Western blot of poly adenosine phosphate ribose polymerase two (PARP) -1 protein expression volume of phosphate buffer. Liquid (PBS) dissolved PBQ, respectively 0,2.5,5.0,10.0,20.0,40.0,80.0160.0 and 320 mol/L PBQ in two groups of cells, cell viability was detected according to the selection of appropriate exposure concentration after application of thiazole blue (MTT) than cell viability assay, Annexin V-PE /7-AAD marked apoptosis flow cytometry detection, real-time fluorescence quantitative polymerase chain the reaction cell PARP-1 gene expression of the two groups. Results the expression of PARP-1 protein in cells of silence (0.15 + 0.02) times and the cells in the control group, (1 + 0.03) times was decreased by 85% compared to.PBQ (P0.05) after 24 h exposure, Significant differences of the two groups of cell viability without the same dose. With the increasing of the dose of PBQ, two groups of cells PARP-1 expression and apoptosis rate were significantly increased (P0.05), and the same dose of PARP-1 silenced cells than the control group apoptosis rate was significantly higher (P0.05). Conclusion PARP-1 silencing BMSCs more susceptible to apoptosis in PBQ under the action of PARP-1 may be involved in PBQ induced apoptosis.

【作者单位】：广东医科大学公共卫生学院劳动卫生与环境卫生教研室;东莞市环境医学重点实验室;广东医科大学公共卫生学院实验中心;
【基金】：广东医科大学科研基金(No.2XB13012)
【分类号】：R329.2
【正文快照】： 1,4-苯醌(PBQ)又称对苯醌,是苯的代谢终产物,在有机合成工业中广泛应用。PBQ作用于骨髓,能引起造血功能损害并导致白血病,但其毒性作用机制尚不完全清楚〔1〕。PBQ在体内能通过形成多种DNA加合物诱导遗传突变的产生,也能够促进或抑制细胞凋亡〔2,3〕。聚腺苷二磷酸核糖聚合酶-

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