吸入氢气对脓毒症小鼠海马组织DNA甲基化的影响  

Effects of inhaled hydrogen gas on the DNA methylation in hippocampus of septic mice

作　　者：于明懂[1] 李佩 于泳浩[3] 卢悦淳[1] 陈会敏 王新[3] 谢克亮[3] YU Ming-dong;LI Pei;YU Yong-hao;LU Yue-chun;CHEN Hui-min;WANG Xin;XIE Ke-liang(Department of Anesthesiology,the Second Hospital of Tianjin Medical University,Tianjin 300211,China;Department of Anesthesiology,Tianjin Hospital;Department of Anesthesiology,Tianjin Medical University General Hospital,Tianjin Research Institute of Anesthesiology)

机构地区：[1]天津医科大学第二医院麻醉科,300211 [2]天津市天津医院麻醉科 [3]天津医科大学总医院麻醉科/天津市麻醉学研究所

出　　处：《天津医药》2020年第3期177-181,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(81671888,81772043);天津市自然科学基金资助项目(18JCZDJC35100)。

摘　　要：目的评价吸入氢气对脓毒症小鼠海马组织的DNA甲基化状态的影响.方法54只健康雄性C57BL/6小鼠采用随机数字表法分为3组:假手术组(Sham组)、脓毒症组(Sepsis组)和氢气治疗组(Sepsis+H2组),每组18只.Sepsis组和Sepsis+H2组采用盲肠结扎穿孔法(CLP)制备小鼠脓毒症模型,Sepsis+H2组小鼠于手术后1 h和6 h吸入用空气混合的2%氢气1 h,Sham组只开腹不进行盲肠结扎和穿孔.3组小鼠于假手术或CLP后1、3、7 d取小鼠海马组织,比色法测定全基因组DNA甲基化水平;实时定量PCR法检测DNA甲基化转移酶(DNMTs,包括DNMT1、DNMT3a和DNMT3b)的mRNA水平;Western blot法检测DNMT1、DNMT3a和DNMT3b蛋白表达水平.结果与Sham组比较,Sepsis组小鼠在建模后1、3、7 d海马组织全基因组甲基化水平明显下降(P<0.05),DNMT1和DNMT3a的mRNA和蛋白表达水平升高,DNMT3b的mRNA和蛋白表达水平降低(P<0.05);与Sepsis组比较,Sepsis+H2组全基因组甲基化水平升高(P<0.05),DNMT1和DNMT3a的mRNA和蛋白表达水平下降,DNMT3b的mRNA和蛋白表达水平升高(P<0.05).结论吸入氢气可纠正脓毒症小鼠海马组织的DNA甲基化紊乱状态,改善DNA甲基化紊乱状态是氢气治疗脓毒症相关性脑病的重要机制之一.Objective To evaluate the effect of inhaled hydrogen gas on DNA methylation in hippocampal tissues of septic mice.Methods Fifty-four male C57BL/6 mice were divided into 3 groups using a random number table:sham operation group(group Sham),sepsis group(group Sepsis)and hydrogen gas group(group Sepsis+H2),18 mice for each group.Sepsis model was produced by cecum ligation and puncture(CLP)in group Sepsis and group Sepsis+H2.Mice in group Sepsis+H2 inhaled 2%hydrogen gas mixed with air at 1 h and 6 h after surgery.Mice in group Sham were performed only laparotomy without CLP.At 1,3 and 7 d after sham operation or CLP,hippocampal tissues were taken from the mice in the three groups,and the global DNA methylation was measured by colorimetry.Real-time PCR was used to detect the mRNA expression levels of DNA methyltransferase enzymes(DNMT1,DNMT3a,and DNMT3b).Western blot assay was used to detect the protein expression levels of DNMT1,DNMT3a and DNMT3b.Results Compared with group Sham,the global DNA methylation in hippocampal tissues was significantly decreased at 1,3 and 7 d after operation in group Sepsis(P<0.05),the mRNA and protein expression levels of DNMT1 and DNMT3a were significantly increased,while the level of DNMT3b was decreased in group Sepsis(P<0.05).Compared with group Sepsis,the global DNA methylation was significantly increased in group Sepsis+H2(P<0.05),the mRNA and protein expression levels of DNMT1 and DNMT3a were significantly decreased,and the level of DNMT3b was significantly increased in group Sepsis+H2(P<0.05).Conclusion Inhaled hydrogen gas could corrected DNA methylation disorders in the hippocampus of septic mice.The correction of DNA methylation disorders is one of the mechanisms of hydrogen gas therapy for sepsis-associated encephalopathy.

关 键 词：氢 脓毒症 DNA甲基化 海马 

分 类 号：R631.2[医药卫生—外科学]