氢醌对人骨髓单个核细胞组蛋白去乙酰化酶的影响

Effect of hydroquinone on the histone deacetylase in human bone marrow mononuclear cells

目的观察氢醌对人骨髓单个核细胞组蛋白去乙酰化酶（HDAC）活力、HDAC1和HDAC2 mRNA表达水平影响，以及加入HDAC抑制剂曲古抑菌素（TSA）作用10h，HDAC活力、HDAC1和HDAC2 mRNA表达水平变化。方法采集正常人骨髓制备单个核细胞悬液，分0h对照组、氢醌3h组、氢醌6h组、氢醌12h组、氢醌24h组、氢醌＋曲古抑菌素10h组、氢醌10h组，收集各组骨髓单个核细胞，提取细胞核蛋白和RNA，应用去HDAC试剂盒检测酶活力变化，实时荧光定量PCR检测HDAC1和HDAC2 mRNA表达水平。结果氢醌3、6、12h组HDAC酶活力分别为0h对照组的1．31、1．53、1．148倍，差异有统计学意义（P〈0．05）。RT-PCR结果显示，除了氢醌24h组，氢醌3、6、12h组HDAC1 mRNA表达量分别为0h对照组的1．173、1．901、2．348倍，差异有统计学意义（P〈0．05）。与对照组比较，氢醌3h组和氢醌24h组HDAC2 mRNA表达量的差异无统计学意义（P〉0．05），氢醌6h组和氢醌12h组的HDAC2 mRNA表达量分别为0h对照组的1．426、1．766倍，差异有统计学意义（P〈0.05）。HQ与TSA作用骨髓单个核细胞10h结果显示，与氢醌10h组比较，氢醌＋曲古抑菌素10h组的HDAC活力降低25．6％，差异有统计学意义（P〈0．05）；与对照组比较，氢醌＋曲古抑菌素10h组的HDAC活力升高13．0％，差异有统计学意义（P〈0.05）。RT-PCR结果显示，HQ与TSA作用骨髓单个核细胞10h，氢醌＋曲古抑菌素10h组比氢醌10h组的HDAC1 mRNA表达水平下降26．9％，HDAC2 mRNA表达水平下降19．3％，与对照组的差异均有统计学意义（P〈0．05）。氢醌10h组及氢醌＋曲古抑菌素10h组的HDAC2 mRNA和HDAC1 mRNA表达水平均明显高于对照组，差异有统计学意义（P〈0.05）。结论在一定时间范围内，氢醌作用骨髓单个核细胞，HDAC活力，HDAC1和HDAC2 mRNA表达水平增加；HDAC抑制剂TSA可以降低氢醌引起的HDAC活力以及HDAC1和HDAC2 mRNA表达水平；提示氢醌引起的骨髓损伤与组蛋白乙酰化修饰水平改变有关。

Objective -To observe the activity of histone deacetylase and the mRNA expression level of HDAC1 and HDAC2 in human bone marrow mononuclear cells ,which induced by hydroquinone and exposed to hydroquinone plus Trichostatin as a histone deacetylase inhibitor for 10 hours respectively. Methods Collect the bone marrow mononuclear cells suspension,divided into control group,HQ group （3 h, 6 h, 12 h, 24 h）, HQ＋ TSA 10 h group and HQ 10 h group. Extract the nuclear proteins and RNA, test the activity of histone deacetylase with the colorimetric HDAC assay kit and detect the mRNA expression level of HDAC 1 and HDAC2 by real-time Polymerase Chain Reaction（PCR）. Results The HDAC activity of HQ3 h group, HQ6 h group and HQ12 h group were 1.31 times, 1.53 times and 1.148 times than that of control group respectively. And the difference was statistically significant （P〈0.05）. Except the HQ24 h group （P〉0.05）, the HDAC1 mRNA expression of HQ3 h group, HQ6 h group and HQ12 h group were 1.173 times, 1.901 times and 2.348 times than that of control group respectively. And the difference was statistically significant （P〈0.05）. The HDAC2 mRNA expression of HQ6 h group and HQ12 h group were 1.426 times and 1.766 times than that of the control group respectively. And the difference was statistically significant （P〈0.05）. No significant difference was observed between HQ3 h group, HQ24 h group and control group（P〉0.05 ）. The cells were treated by hydroquinone plus TSA for 10 hours. The HDAC activity of HQ＋TSA 10h group was reduced by 25.6% than that of HQ 10 h group （P〈0.05） and rised 13.0% compared to the control group （P〈0.05）. And the difference was statistically significant between groups（P〈0.05 ）.The cells were treated by hydroquinone plus TSA for 10 hours. The HDAC1 mRNA expression of the HQ＋TSA 10h group is reduced by 26.9% than that of HQ10h group. The HDAC2 mRNA expression is reduced by 19.3% compared to the HQ 10h group.And the difference was statistically significant between groups （P〈0.05）. The HDAC1 and HDAC2 mRNA expression is obviously higher than the control group , the difference was statistically significant （P〈0.05）. Conclusion Treatment of hydroquinone, the histone deaeetylase activity and the mRNA expression of HDAC1 and HDAC2 were increased in a certain time range. The histone deacetylase inhibitor （TSA） can reduce the histone deacetylase activity and the mRNA expression level of HDAC1 and HDAC2 in the bone marrow mononuclear cell induce by hydroquinone.h can be confirmed that hematopoietie damage induced by the benzene metabolites is related to the histone aeetylation modification level.