中文摘要：

目的探讨沉默聚腺苷二磷酸核糖聚合酶-1（PARP-1）对氢醌所致大鼠骨髓间充质干细胞（BMMSCs）凋亡的影响。方法①PARP-1基因RNA干扰表达质粒转染BMMSCs后,经新霉素筛选获得PARP-1沉默BMMSCs,以免疫印迹法鉴定所构建的细胞。②以空载体BMMSCs为对照组,以PARP-1沉默BMMSCs为实验组。分别以质量浓度为0.0、2.5、5.0、10.0、20.0、40.0、80.0、160.0和320.0μmol/L的氢醌（溶于磷酸盐缓冲液）作用于2组细胞24 h,以噻唑蓝法检测细胞活力,根据细胞活力检测结果选择后续实验氢醌染毒剂量。③以浓度为0.0-20.0μmol/L的氢醌作用于2组细胞24 h后,以流式细胞技术检测细胞凋亡情况,以实时荧光定量-聚合酶链反应检测PARP-1 mRNA表达情况。结果①用质量浓度为400 mg/L的新霉素成功筛选出PARP-1沉默BMMSCs和空载体BMMSCs,PARP-1沉默BMMSCs中PARP-1蛋白表达量较BMMSCs下降85.00%,成功建立PARP-1沉默细胞。②根据细胞活力检测结果,后续实验选择氢醌染毒剂量为0.0-20.0μmol/L。3与氢醌染毒剂量为0.0μmol/L时比较,对照组及实验组细胞早期凋亡率分别在氢醌染毒剂量为10.0和5.0μmol/L时即出现有统计学意义的升高（P〈0.05）;且在浓度为0.0-10.0μmol/L时,随着氢醌染毒剂量的增加,2组细胞早期凋亡率均增加（P〈0.01）,均呈剂量-效应关系。③与氢醌染毒剂量为0.0μmol/L比较,对照组和实验组细胞的PARP-1 mRNA相对表达水平均在氢醌染毒剂量为5.0μmol/L时即出现有统计学意义的升高（P〈0.05）,且在每个氢醌染毒剂量下实验组细胞PARP-1 mRNA相对表达水平均低于对照组（P〈0.05）;在浓度为0.0-20.0μmol/L时,随着氢醌染毒剂量的增加,2组细胞PARP-1 mRNA相对表达水平均增加（P〈0.01）,均呈剂量-效应关系。结论 PARP-1沉默BMMSCs在氢醌作用下更易发生凋亡,PARP-1可能参与了氢醌诱导的细胞凋亡。

英文摘要：

Objective To explore the effect of silencing of poly（ ADP-ribose） polymerase-1（ PARP-1） on cell apoptosis induced by hydroquinone（ HQ） in rat bone marrow-derived mesenchymal stem cells（ BMMSCs）. Methods i） The RNA expression vectors for PARP-1 gene were transfected into BMMSCs. Neomycin was used to select the transfected cells that stably expressed PARP-1-shRNA. Western blotting was used to examine the gene silencing efficiency. ii） BMMSCs with PARP-1 silencing were sorted as the treated group,while BMMSCs with empty vector were considered as the control group.HQ dissolved in phosphate buffer solution at the concentrations of 0. 0,2. 5,5. 0,10. 0,20. 0,40. 0,80. 0,160. 0 and320. 0 μmol / L were given to both groups for 24 hours. The cell viability was detected by methyl thiazolyl tetrazolium assay,and the concentration of HQ was chosen for the following study based on cell viability. iii） Both groups were treated by HQ at concentrations of 0. 0-20. 0 μmol / L for 24 hours,then the apoptosis of BMMSCs was detected by flow cytometry. The PARP-1 mRNA expression was determined by real-time fluorescent quantitative polymerase chain reaction assay. Results i） PARP-1 silencing cells and empty vector control cells were successfully screened with a mass concentration of 400 mg / L neomycin,and confirmed by level of protein expression. The interference efficiency of PARP-1 gene and inhibition efficiency was 85. 00%. ii） Based on the result of cell viability,HQ at concentrations of 0. 0-20. 0 μmol / L was chosen for the following study. iii） Compared with the group treated by HQ at concentration of 0. 0 μmol / L,the rate of early apoptosis of control group increased significantly with HQ at concentration of 10. 0 μmol / L while that of treated group was increased significantly at concentration of 5. 0 μmol / L（ P〈0. 05）. In addition,at concentrations of 0. 0-10. 0 μmol / L,the rates of early apoptosis in both groups increased in a dose-dependent manner（ P〈0. 01）. Compared with