氢气对重度脓毒症小鼠肠道损伤的保护作用

Protective effects of hydrogen on intestinal injury in septic mice

目的观察氢气吸入对重度脓毒症小鼠肠黏膜屏障功能、肠道组织氧化还原状态的影响,并探讨其机制。方法选取美国癌症研究所雄性小鼠56只,体质量20~25 g,均为5周龄,采用随机数字表法将其分为四组：假手术组（A组）、假手术＋氢气组（B组）、脓毒症组（C组）和脓毒症＋氢气组（D组）,每组14只。采用盲肠结扎穿孔（CLP）法制备小鼠脓毒症模型。B组和D组于CLP后1、6 h吸入2%氢气1 h。CLP后24 h进行行为学评分;采集静脉血检测血中生化指标[丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、尿素氮（BUN）、肌酐（Cr）];随后处死小鼠取中段空肠,对肠道组织进行原位末端脱氧核苷酸转移酶标记法（TUNEL）染色,观察肠道组织细胞凋亡情况并评分;检测血液及肠道组织匀浆中丙二醛（MDA）水平及过氧化氢酶（CAT）、超氧化物歧化酶（SOD）的活力。结果与A组比较,C组和D组行为学评分、肠道组织凋亡指数、血生化指标（ALT、AST、BUN、Cr）水平及血清和肠道组织MDA水平升高,SOD和CAT活力降低,差异均有统计学意义（P〈0.05）,而B组上述指标与A组比较,差异均无统计学意义（P〉0.05）;与C组比较,D组行为学评分、肠道组织凋亡指数、血生化指标水平及血清和肠道组织MDA水平降低,SOD和CAT活力升高,差异均有统计学意义（P〈0.05）。结论氢气吸入对重度脓毒症小鼠肠损伤和肠黏膜屏障功能破坏有明显保护作用,可能与其抗凋亡作用及对内源性氧化还原平衡状态的维持有关。

Objective To investigate the effect of hydrogen（H2） inhalation on the intestinal barrier function and intesti nal tissue redox status in severe septic mice and its mechanism. Methods Fifty-six male mice of the American Institute for Cancer Research（AICR） were selected,the body mass was 20-25 g and aged 5 weeks,and divided into four groups according to the random number table method ： sham operation group（ group A）, sham operation ＋ H2group（ group B）, sepsis group（ group C）and sepsis＋H2group（group D）,14 cases in each group. The mouse sepsis model was produced by cecal ligation and puncture（CLP）method. The group B and D received 1 h inhalation of 2% H2 at 1 and 6 h after CLP. The behavioral score was conducted at 24 h after CLP;the venous blood was collected for detecting blood biochemical indicators（ALT,AST,BUN,Cr）,then the mice were sacrificed for taking the middle section of jejunum. The intestinal tissue was performed the TUNEL staining for observing the apoptosis situation of intestinal tissue cells and conducting the scoring;the levels of MDA and activity of CAT and SOD in blood and intesti nal tissue homogenate were detected. Results Compared with the group A,the behavioral score,intestinal tissue apoptosis index,levels of blood biochemical indicators（ALT,AST,BUN,Cr） and serum and intestinal tissue MDA level in the group C and D were increased,while the activity of SOD and CAT were decreased,the differences were statistically significant（P〈0.05）,but which in the group B had no statistical differences as compared with the group A（P〉0.05）;compared with the group C,the behavioral score,intestinal tissue apoptosis index,levels of blood biochemical indicators and serum and intestinal tissue MDA level in the group D were decreased,while the activity of SOD and CAT were increased,the differences were statistically significant（P〈0.05）.Conclusion H2 inhalation has obviously protective effect on intestinal injury and intestinal mucosal barrier function destroy in severe septic mice,which may be associated with the anti-apoptotic effect and maintaining endogenous redox equilibrium state.