摘要
目的探讨聚腺苷二磷酸核糖聚合酶-1(PARP-1)特异性抑制剂N-(6-氧-5,6-二氢菲啶-2-羟基)-N,N-二甲基乙酰胺-盐酸(PJ34)对氢醌(HQ)所致人胚肺成纤维细胞(HLF细胞)凋亡的影响及其作用机制。方法将离体传代培养的HLF细胞分为阴性对照组、阳性对照组(HQ组)、3个PJ34组和3个PJ34+HQ组。按PJ34不同浓度(0.5、1.0、2.0μmol/L)分为PJ34的Ⅰ、Ⅱ、Ⅲ组,作用4 h后检测结果;按PJ34的Ⅰ、Ⅱ、Ⅲ组浓度及作用4 h后再分别加入HQ(80μmol/L)分为PJ34+HQ的Ⅰ、Ⅱ、Ⅲ组,作用24 h检测结果;HQ组仅用HQ24 h处理。采用PARP-1单克隆抗体荧光标记,流式细胞术检测HLF细胞中PARP-1蛋白表达、细胞凋亡和细胞复制后期(G2)细胞数情况,噻唑蓝(MTT)比色法测定HLF细胞相对存活率。结果各组给予不同剂量的PJ34 4 h后,HLF细胞的存活(增殖)明显受抑制(P<0.05),随着PJ34浓度的增加,其抑制作用进一步增强;PARP-1蛋白表达阳性的HLF细胞百分数均明显低于阴性对照组(P<0.01);流式细胞术检测显示HQ作用24 h后均可引起HLF细胞的调亡,可出现明显的亚二倍体峰,且随着PJ34预处理浓度的增加,其峰值也逐渐增加;HQ可导致细胞周期出现G2期阻滞,PJ34+HQⅠ、Ⅱ、Ⅲ组G2期阻滞减少且差异具有显著性(P<0.05或P<0.01)。结论PARP-l抑制剂PJ34明显抑制HLF细胞PARP-1表达,增加HQ所致细胞凋亡,并通过诱导HLF细胞凋亡抑制其增殖。
Objective To study the changes of apoptosis and its mechanism of PJ34 of poly （ADP-fibose）polymerase-1 （PARP-1） inhibitor on human embryo lung fibroblasts （HLF） cells induced by hydroquinone （HQ） in vitro. Methods HLF cells were randomly divided into a control group, an HQ control group and test groups. HLF cells in test groups were pretreated with PJ34 at the concentrations of 0.5, 1.0, 2.0 μmol./L （PJ34 groups）, and 4 hours later, 80 μmol/L of HQ （PJ34 ＋HQ groups） were added in respectively. Monoclonal antibody labeled by fluorescenee（FITC） and Flow cytometry （FCM） were used to examine the PARP-1 expression of the controls and the apoptosis rate in the PJ34-pretreated and HQ-treated HLF ceils. Resuits PJ34 suppressed the growth of HLF cells （P〈0.05）, the suppressive effects were enhanced with the increase of PJ34 concentration. The percentage of HLF cells with positive expressions of PARP-1 protein of PJ34 groups was significantly decreased compared with that of the control group（P〈0.01）. FCM showed that the changes of apoptosis rates of HLF ceils treated with HQ for 24 hours. In addition, a typical subdiploid peak could distinctively appear, the peak value was increased with the increase of PJ34 concentration. G2 phase of cell cycle arrest induced by HQ in vitro, and G2 phase arrest decreased in PJ34 ＋ HQ groups （P 〈 0. 01 or P 〈 0. 05）. Conclusion PJ34 PARP-1 inhibitor could rapidly inhibit the PARP-1 expression of HLF ceiLs, enhance the apoptosis, and directly inhibit the cell growth of HLF cells through inducing cell apoptosis.
出处
《中国职业医学》
CAS
北大核心
2008年第6期451-453,457,共4页
China Occupational Medicine
基金
广东省自然科学基金(7301507)
广东省医学科研基金资助课题(A2005471)
广东医学院面上项目(XK0407)
关键词
聚腺苷二磷酸核糖聚合酶-1
PJ34
氢醌
凋亡
人胚肺成纤维细胞
Poly （ADP-ribose） polymerase-1 （ PARP-1 ）
PJ34
Hydroquinone （HQ）
Apoptosis
Human embryo lung fibroblasts （HLF）
