Repair effect of 2%hydrogen inhalation on neurological function in rats with sepsis-related encephalopathy and its mechanism
Objective To observe the effect of 2%hydrogen inhalation on the repair of neurological function in rats with sepsis-associated encephalopathy and to explore its possible mechanism.Methods Thirty-six SD rats were random-ly divided into the group S,group H,and group SH,with 12 rats in each group.Rats in the groups S and H were used to establish sepsis models by cecal ligation and puncture,rats in the group SH only underwent laparotomy without cecal liga-tion,rats in the group H inhaled 2%hydrogen for 24 h after successful modeling,and rats in the groups S and SH normally inhaled air for 24 h.Neurobehavioral scores were performed at 0,6,12,and 24 h after inhalation in each group,and four rats were randomly selected and sacrificed at 6,12,and 24 h after inhalation.HE staining was used to observe the patho-logical changes of brain tissues.The expression of S100β and Caspase-1 was detected by Western blotting and immunohis-tochemistry.HT22 cells in logarithmic growth phase were divided into the air group,hydrogen group and control group.The cell sepsis injury models were established in the air and hydrogen groups,and lipopolysaccharide was added to the cul-ture medium.The hydrogen group was placed in a 37℃,2%hydrogen incubator,while the air group was placed in a 37℃,5%CO₂ incubator.The control group was maintained in a 37℃,5%CO₂ incubator with normal culture medium.All groups were cultured for 24 h.TUNEL assay was applied to detect apoptosis rate.The expression of Caspase-1 was de-tected by Western blotting.Results The neurobehavioral scores of rats in both the group S and the group H at 0 h of in-halation were lower than those in the group SH.The neurobehavioral scores decreased successively in the group SH,group H and group S after inhalation for 6,12 and 24 h(both P<0.05).Pathological observation of brain tissues:In the group SH,the neurons were neatly arranged,the structure was clear and the number of neurons was large in the brain tissue after inhalation for 6 h.Over the inhalation time,the number of normal neurons did not change significantly.In the group S,neurons were disorganized,cell nuclei were pyknotic,cell structure was loose,and Nissl bodies in the cytoplasm were re-duced at 6 h of inhalation,and the number of normal neurons gradually decreased over the inhalation time.In the group H,the neurons were arranged neatly at 6 h after inhalation,and the number of normal neurons gradually increased with the prolongation of inhalation time.At 6 and 12 h after inhalation,the positive expression rates of S100β and Caspase-1 pro-tein and the relative expression levels of S100β and Caspase-1 protein in the brain tissues of rats in the groups S,H,and SH decreased in turn,with statistically significant difference between groups(all P<0.05).After 24 hours of inhalation,the positive expression rates of S100β and Caspase-1 protein and the relative expression levels of S100β and caspase-1 pro-tein in the brain tissues of rats in the groups H and SH were lower than those in the group S(all P<0.05).The apoptosis rate and relative expression of Caspase-1 protein in control group,hydrogen group and air group increased in turn,and the relative expression of Caspase-1 protein in cells of both the control group and the hydrogen group was lower than that in the air group(all P<0.05).Conclusion Inhalation of 2%hydrogen promotes neurological recovery in rats with sepsis-asso-ciated encephalopathy,and the mechanism may be related to the inhibition of neuronal pyroptosis.
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