三氧化二砷对MRL/lpr狼疮鼠脾脏淋巴细胞组蛋白乙酰化的影响  

The Effects of Arsenic Trioxide on Acetylation of Histone H3 of Splenic Lymphocytes in MRL/lpr Mice

作　　者：嘉婷[1] 强孚勇[1] 张挺[1] 朱载华[1] 朱小春[1] 

机构地区：[1]温州医学院附属第一医院风湿免疫科,温州325000

出　　处：《中国细胞生物学学报》2010年第5期744-748,共5页Chinese Journal of Cell Biology

基　　金：温州市科技计划(No.Y20090240)资助项目~~

摘　　要：观察三氧化二砷(arsenic trioxide,ATO)对MRL/lpr狼疮鼠脾脏淋巴细胞组蛋白H3乙酰化水平的影响以及对组蛋白乙酰基转移酶(histone acetyltransferases,HATs)和组蛋白去乙酰化酶(histone deacetylases,HDACs)活性的影响。将20周龄MRL/lpr狼疮鼠和正常C57BL/6J小鼠无菌条件下取出脾脏,配制成脾脏淋巴细胞悬液。体外经PHA-P(20μg/ml)和IL-2(1000 U/ml)常规刺激48 h后,随机分为3组:ATO组;曲古抑菌素(Trichostatin A,TSA)组;RPMI 1640组。继续培养24 h。免疫印迹法测定乙酰化组蛋白H3的表达量;酶活性试剂盒分别测定HAT和HDAC活性。结果显示在MRL/lpr狼疮鼠中RPMI 1640组、ATO组、TSA组组蛋白H3乙酰化水平分别为0.46±0.06、O.87±0.02、1.6±0.13(n=3),且差异具有显著的统计学意义(P<0.01);ATO和TSA能明显降低HDAC活性(P<0.01);而对于HAT,三组间两两相比差异无统计学意义;ATO组和TSA组HDAC/HAT的比值明显低于RPMI 1640组。而在C57BL/6J小鼠中,ATO组和RPMI 1640组之间以上指标均无差异。研究结果表明AT0能明显抑制MRL/lpr狼疮鼠脾脏淋巴细胞的HDAC活性,下调HDAC/HAT的比值,提高组蛋白H3的乙酰化水平,而对正常小鼠并无明显影响。This study aimed to investigate the effects of arsenic trioxide（ATO） on the acetylation status of the histone H3 of splenic lymphocytes from MRL/lpr mice and the activities of histone acetyltransferases（HATs） and histone deacetylases（HDACs）.Spleens were isolated from MRL/lpr mice and C57BL/6J mice at 20 weeks of age under specific pathogen-free condition and made to suspensions.Splenic lymphocytes were stimulated in vitro for 48 h in the presence of PHA-p（20μg/ml） and IL-2（1 000 U/ml）,then were divided randomly into 3 groups：ATO group（treated with 1.0μmol/L ATO for 24 h）,TSA group（treated with 300 ng/ml TSA for 24 h） and RPMI 1640 group（treated with RPMI 1640 for 24 h）.Expression levels of acetylated histones H3 in 20-week-old MRL/lpr splenocytes were detected by Western blot.The activities of HDAC and HAT were evaluated via the corresponding activity assay kits.The results showed that in MRL/lpr mice,the degrees of acetylated histone H3 in RPMI 1640 group,ATO group and TSA group were 0.46±0.06,0.87±0.02 and 1.6±0.13,respectively.And the difference was statistically significant（P0.01）.Compared with RPMI 1640 group,ATO group and TSA group could significantly reduce the activity of HDAC（P0.01）.However,for the HAT activity,no significant difference was observed between ATO group as well as between TSA group versus RPMI 1640 group.HDAC/HAT ratios in ATO group as well as TSA group were lower than RPMI 1640 group.There was no difference between ATO group and RPMI 1640 group in C57BL/6J mice（P0.05）.It was concluded that ATO can inhibit the activity of HDAC,lower the ratio of HDAC/HAT,and improve the acetylation level of histone H3 in the MRL/lpr splenic lymphocytes,and it was of no effect in normal mice.

关 键 词：系统性红斑狼疮 三氧化二砷 组蛋白 乙酰化 组蛋白去乙酰化酶 

分 类 号：R593.241[医药卫生—内科学]