表基因修饰参与H_2O_2介导人晶状体上皮细胞SRA01/04的生长停滞  

Growth arrest of human lens epithelial cell SRA01/04 mediated by H_2O_2 conbined with epigenetic modification

作　　者：王新[1] 李宝华[2] 张明昌[1] 张向东[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,湖北省武汉市430022 [2]郑州人民医院,河南省郑州市450003 [3]新乡医学院第三附属医院眼科,河南省新乡市453003

出　　处：《眼科新进展》2013年第8期721-724,共4页Recent Advances in Ophthalmology

摘　　要：目的探讨在H2O2介导人晶状体上皮细胞SRA01/04生长停滞下,表基因修饰的改变。方法将培养的人晶状体上皮细胞SRA01/04分为3组,分别为:A组:先加H2O2组,即在细胞培养液内先加0.5μmol·L-1H2O2,30min后再加吡诺克辛液300μL培养23.5h;B组:后加H2O2组,即先在细胞培养液内加吡诺克辛钠液300μL培养23.5h后,再加0.5μmol·L-1H2O2培养30min;C组:对照组,即不加H2O2及药物培养24h。对各组细胞进行免疫细胞化学检测和Western bolt检测,分析各组细胞内组蛋白脱乙酰化酶(histone deacetylase,HDAC)1、核因子-κB(nuclear factor-KappaB,NF-κB)、c-myc及周期蛋白1(Cyclin D1)的表达。结果免疫细胞化学检测结果显示,NF-κB表达信号的灰度值:A组(192.25±5.56)>B组(170.40±2.50)>C组(157.14±1.09),3组间两两相比差异均有统计学意义(均为P<0.01);HDAC1、c-myc及Cyclin D1表达信号的灰度值均为C组>B组>A组,且3组间两两相比差异均有统计学意义(均为P<0.01)。Western blot检测结果显示,相较于对照组,以H2O2处理人晶状体上皮细胞SRA01/04后HDAC1、Cyclin表达减弱,并且A组比B组更为明显;同时可见NF-κB的表达增强,A组也比B组更为明显。结论在H2O2介导人晶状体上皮细胞SRA01/04生长停滞下,HDAC1活性减弱,NF-κB激活,c-myc及Cyclin D1的表达下调。Objective To explore the epigenetic modification change under the growth arrest of human lens epithelial cell SRA01/04 mediated by H202. Methods The cultured human lens epithelial cells SRA01/04 were divided into 3 groups：Group A or pre-H202 group,that is after 0.5 μmol · L-1 H202 was added to the cultured SRA01/ 04 cells for 30 minutes,the cells were co-cultivated with 300μL Pirenoxine solution for 23.5 hours;Group B or post-H202 group,that is after co-culture of the SRA01/04 cells with 300μL Pirenoxine solution for 23.5 hours,0.5 μmol · L-1 H202 was added and continued to cultivate for 30 minute; Group C or control group, that is SRA01/04 cells were cultured for 24 hours with neither H2 02 nor drug. Each group was detected by im- munocytochemical reaction and Western bolt and the expression of histone deacetylase （HDAC） 1, nuclear factor-Kappa B （NF-KB）, c-myc and Cyclin D1 was analyzed. Re- suits The immunocytochemical reaction showed that the scanned gray-scaled means of NF-KB in group A（ 192.25 ± 5.56） 〉 group B（ 170.40 ± 2.50） 〉 group C（ 157. 14 ± 1.09）. The three groups all had statistical（ all P 〈 0.01 ） ; Those of HDAC1, c-myc and CyclinD1 were all group C 〉 group B 〉 group A and these groups all had statistical sig- nificance（ all P 〈 0. 01 ）. The Western bolt showed that compared with control group, HDAC1 activity decreased after treated with n202 and group A appeared more clearly than group B;The expression of NF-KB became stronger, which appeared more clearly in group A than that in group B. Conclusion Under the growth arrest of human lens epithelial cell SRA01/04 mediated by H202 ,the HDAC1 activity decreases,NF-KB factor is up-regulated and the expressions of c-myc and CyclinD1 are down ragulated.

关 键 词：氧化应激 晶状体上皮细胞 组蛋白脱乙酰化 

分 类 号：R776.1[医药卫生—眼科]