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
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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、40 μmol/L,继续培养48 h;第二批分5 个组,以20 μmol/L B(a)P 染毒,分别于染毒0、6、12、24、48 h 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0 h 组相比,染毒48 h 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。 Abstract: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 0 h(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 HDAC2 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.
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