氢对脓毒症相关性脑病小鼠脑组织线粒体功能的影响

Effect of hydrogen on mitochondrial function in brain tissues of mice with sepsis-associated encephalopathy

目的评价氢对脓毒症相关性脑病小鼠脑组织线粒体功能的影响。方法健康雄性C57小鼠268只,体重20~25 g,6周龄,采用随机数字表法将其分为4组(n=67):假手术组(SH组)、假手术+氢气组(SH+H2组)、脓毒症相关性脑病组(SAE组)和脓毒症相关性脑病+氢气组(SAE+H2组)。采用盲肠结扎穿孔术制备脓毒症相关性脑病模型。SH+H2组和SAE+H2组分别于术后1、6 h时吸入2%氢气1 h。记录术后7生存情况。于术后24 h时取脑组织,计数海马CA1区正常神经元;于术后6、12和24 h时,采用荧光分光光度法检测海马线粒体膜电位(MMP),采用荧光素-荧光酶发光法检测线粒体ATP含量;于术后3、5和7 d时行Y迷宫自发交替实验。结果与SH组比较,SAE组和SAE+H2组7 d生存率、海马CA1区正常神经元计数、MMP、线粒体ATP含量和自发交替百分比降低(P<0.05);与SAE组比较,SAE+H2组7 d生存率、海马CA1区正常神经元计数、MMP、线粒体ATP含量和自发交替百分比升高(P<0.05)。结论氢减轻小鼠脓毒症相关性脑病的机制可能与改善脑组织线粒体功能有关。

Objective To evaluate the effect of hydrogen on the mitochondrial function in brain tissues of mice with sepsis-associated encephalopathy(SAE).Methods Two hundred and sixty-eight healthy male C57 mice,aged 6 weeks,weighing 20-25 g,were divided into 4 groups(n=67 each)using a random number table method:sham operation group(group SH),sham operation plus hydrogen gas group(group SH+H2),group SAE,and SAE plus hydrogen gas group(group SAE+H2).Sepsis was produced by cecal ligation and puncture in anesthetized mice.The mice in group SH+H2 and group SAE+H2 inhaled 2%hydrogen gas for 1 h starting from 1 and 6 h after operation,respectively.The postoperative 7-day survival rate was recorded.Brain tissues were obtained at 24 h after operation to count the normal neurons in hippocampal CA1 region.At 6,12 and 24 h after operation,hippocampal mitochondria were isolated for determination of mitochondrial membrane potential(MMP)by fluorescence spectrophotometry and ATP content by a bioluminescence assay.Y-maze(spontaneous alternation)test was performed at days 3,5 and 7 after operation.Results Compared with group SH,the 7-day survival rate was significantly decreased,the number of normal neurons in hippocampal CA1 region,MMP,mitochondrial ATP content and spontaneous alternation percentage in Y-maze test were significantly decreased in group SAE and group SAE+H2(P<0.05).Compared with group SAE,the 7-day survival rate,the number of normal neurons in hippocampal CA1 region,MMP,mitochondrial ATP content and spontaneous alternation percentage in Y-maze test were significantly increased in group SAE+H2(P<0.05).Conclusion The mechanism by which hydrogen reduces SAE is probably associated with improving mitochondrial function in brain tissues of mice.