天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (3): 177-181.doi: 10.11958/20192608

• 实验研究 • 上一篇    下一篇

吸入氢气对脓毒症小鼠海马组织 DNA甲基化的影响

于明懂，李佩，于泳浩，卢悦淳,陈会敏，王新，谢克亮   

  1. 1天津医科大学第二医院麻醉科（邮编 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
  • 收稿日期:2019-08-23 修回日期:2020-01-10 出版日期:2020-03-15 发布日期:2020-04-11
  • 通讯作者: 于明懂 E-mail:ymd283640731@163.com

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

YU Ming-dong,LI Pei,YU Yong-hao,LU Yue-chun,CHEN Hui-min,WANG Xin,XIE Ke-liang   

  1. 1 Department of Anesthesiology, the Second Hospital of Tianjin Medical University, Tianjin 300211, China; 2 Department of Anesthesiology, Tianjin Hospital; 3 Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin Research Institute of Anesthesiology
  • Received:2019-08-23 Revised:2020-01-10 Published:2020-03-15 Online:2020-04-11
  • Contact: YU MingDong E-mail:ymd283640731@163.com

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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。

关键词: 氢, 脓毒症, DNA甲基化, 海马

Abstract: 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.

Key words: hydrogen, sepsis, DNA methylation, hippocampus