Abstract:

Objective To investigate the effect of hydrogen on myocardiumapoptosis after limb ischemia-reperfusion in rabbits through inhibition of the tumor necrosis factor-like cytokine 1A （TL1A）/death receptor 3 （DR3） pathway. Methods The gene expression omnibus （GEO） GSE160516 dataset was downloaded and the kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment was performed. Eighteen male New Zealand large-eared white rabbits were randomly divided into control group （Group C）， limb ischemia-reperfusion group （Group R）， and hydrogen group （Group H） using the random number table method， with six rabbits in each group. The rabbit double hind limb ischemia-reperfusion model was established， and the animals in each group were centrifuged with venous blood to prepare serum， and the hearts were removed. Serum TL1A and DR3 levels were measured by enzyme-linked immunosorbent assay （ELISA）. The terminal deoxynucleotidyl transferase UTP mediated nicked end labeling （TUNEL） assay was performed to detect apoptosis rate in myocardium. The relative expression of TL1A， DR3 and cysteine aspartate protease 3 （Caspase3） proteins in myocardium was measured by western blotting （WB）. The real-time reverse transcription polymerase chain reaction （RT-qPCR） was performed to determine the relative expression of TL1A， DR3 and Caspase3 mRNA in myocardium. Results KEGG enrichment analysis revealed that gene expression was enriched to Apoptosis signaling pathway，TNF signaling pathway and MAPK signaling pathwayafter myocardium ischemia-reperfusion in mice. It was observed that the expression levels of serum TL1A and DR3 were significantly higher in group R compared with group C （P < 0.01）， while the expression levels of serum TL1A and DR3 were significantly lower in group H compared with group R （P < 0.01）. Compared with group C， the apoptosis rate of myocardium in group R was significantly higher （P < 0.01）；compared with group R， the apoptosis rate of myocardium in group H was significantly lower （P < 0.01）. The relative expressions of TL1A， DR3 and Caspase3 protein and mRNA in myocardium were significantly increased in group R compared with group C （all P<0.01）； the relative expressions of TL1A， DR3 and Caspase3 protein and mRNA in myocardium were significantly decreased in group H compared with group R （P < 0.05， P < 0.01）. Conclusion Hydrogen attenuates myocardium apoptosis after limb ischemia-reperfusion by inhibiting the TL1A/DR3 pathway.

Key words: hydrogen, ischemia-reperfusion, myocardium, apoptosis