组蛋白去乙酰化酶2基因在苯并(a)芘致神经细胞凋亡中的量效和时效表达  

The Dose-effect and Time-effect Expression of Histone Deacetylase 2 mRNA in B(a)P-induced Neural Cell Apoptosis

作　　者：王晓燕[1] 李娜[1] 牛侨[1] 聂继盛[1] 

机构地区：[1]山西医科大学公共卫生学院劳动卫生教研室,山西太原030001

出　　处：《环境与职业医学》2011年第4期219-223,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金资助项目(编号:30800899;30471437);山砥省自然科学基金(编号:2010021034-3)

摘　　要：[目的]通过体外培养细胞途径研究组蛋白去乙酰化酶2(HDAC2)基因在苯并(a)芘[B(a)P]所致神经细胞凋亡中的量效和时效表达。[方法]大鼠原代培养的神经元细胞分为两批:第一批分4个组,以B(a)P同时加入S9对细胞染毒,分别为二甲基亚砜(DMSO)组、低剂量组、中剂量组、高剂量组,使其终浓度为0、10、20、40μmol/L,继续培养48h;第二批分5个组,以20μmol/L B(a)P染毒,分别于染毒0、6、12、24、48h处理细胞。用流式细胞术检测神经细胞的凋亡率,实时荧光定量PCR法检测caspase-3,HDAC2基因的量效和时效表达情况。[结果]①流式细胞术结果显示,与DMSO组相比,中、高剂量组神经细胞的凋亡率明显增加(P<0.05);与低剂量组相比,高剂量组凋亡率增加(P<0.05)。与0 h组相比,染毒24、48 h组神经细胞的凋亡率明显增加(P<0.05)。②与DMSO组相比,高剂量组caspase-3、HDAC2mRNA的表达量明显增加(P<0.01);与低剂量组相比,高剂量组caspase-3 mRNA表达量明显增加(P<0.05)。与0h组相比,染毒48h组caspase-3、HDAC2 mRNA表达量明显升高(P<0.01,P<0.05)。[结论]B(a)P可引起神经细胞凋亡,HDAC2基因表达上升,该基因可能在B(a)P所导致的神经细胞凋亡中起重要作用。[ Objective ] To investigate the dose-effect and time-effect expression of histone deacetylase 2 （ HDAC2 ） mRNA in B（a）P-induced neural cell apoptosis in vitro. [ Methods ] The primary cultured neurons of rat were assigned into two lots. The first lot was separated into 4 groups and treated with B（a）P at final concentrations as 0 μmol/L（ DMSO group ）, 10 μmol/L （ low-dose group ）, 20 μmol/L （ medium-dose group ）, and 40 μmol/L （ high-dose group ） respectively, and added S9 at the same time, then incubated for 48 hours. The second lot was separated into 5 groups, all treated with 20 μmol/L B（a）P, then incubated for 0, 6, 12, 24, 48 h respectively. The apoptosis rate of all incubated neurons was detected by flow cytometry, the expression of caspase-3 and HDAC2 mRNA by QRT-PCR. [ Results ] （1）Flow cytometry showed that compared with the DMSO group, the apoptosis rates of neurons in middle and high-dose group were significantly increased （ P 〈 0.05 ） ; compared with the low-dose group, the apoptosis rates also increased （P〈 0.05 ）. The apoptosis rates at 24, 48 h after exposure were significantly increased compared with that at 0h（P〈0.05 ）. （2）The caspase-3 and HDAC2 mRNA expression showed significant increments in highdose group compared with DMSO group（ P 〈 0.01 ）. The caspase.3 mRNA expression in high dose group increased significantly compared with the low-dose group （ P 〈 0.05 ）. The expressions of caspase-3 and HDA C2 mRNA at 48 h after exposure increased significantly compared with that at 0 h （P 〈 0.01, P 〈 0.05 ）. [ Conclusion ] After exposed to B（a）P, neural cell apoptosis and HDAC2 mRNA expression increased. HDAC2 may play an important role in B（a）P-induced neuronal apoptosis.

关 键 词：苯并(A)芘 组蛋白去乙酰化酶2 半胱天冬酶-3 细胞凋亡 

分 类 号：R114[医药卫生—卫生毒理学]