Abstract: Abstract：Objective To investigate the effect of miR-182-5p targeting FoxO3a-mediated pyroptosis on hepatic ischemia-reperfusion (IR) injury. Methods Liver ischemia-reperfusion models of mice were established. According to the random number table method, 40 mice were divided into 5 groups with 8 rats in each group respectively, sham group, IR groups (2 h group, 6 h group, 12 h group and 24 h group). The cell experiments were divided into two parts: (1) the hypoxia models were established and divided into control group and IR group. (2) Hypoxia/reoxygenation models were established and divided into control group, mimic group, inhibitor group and inhibitor+siRNA group. HE staining was used to observe the pathological changes of liver tissues after IR. Immunocytochemistry staining was used to detect the expression of FoxO3a in liver cells. qRT-PCR and Western blot assay were carried out to detect the expressions of miR-182-5p, FoxO3a, Caspase1, IL-1β and IL-18. The gene correlation between miR-182-5p and FoxO3a was analyzed. Immunofluorescent staining was used to detect the Caspase1 expression. ELISA kit was used to detect the expression of IL-1β and IL-18. CCK- 8 kit was used to detect changes of cell viability. Results The liver injury after IR treatment was increased in mice, and the injury was the most severe at 12 h-reperfusion. At the same time, the expression levels of FoxO3a, Caspase1, IL-1β and IL- 18 were increased (P＜0.05), and pyroptosis was induced. MiR-182-5p level in liver tissue was higher after IR treatment compared with that in sham group (P＜0.05). After IR treatment for AML12 cells in vitro, miR-182-5p expression was up regulated, FoxO3a expression was down-regulated, and caspase1 expression was increased (P＜0.05). Overexpression of miR-182-5p decreased the expression of FoxO3a. On the contrary, it increased the expression of FoxO3a, which in turn decreased the expression levels of Caspase1, IL-1β and IL-18. In the end, the viabilities of hepatocytes were increased (P＜ 0.05). Conclusion The activation of miR-182-5p can aggravate hepatic ischemia-reperfusion injury, while the inhibition of miR-182-5p can reduce hepatic ischemia-reperfusion injury. The mechanism may be related to miR-182-5p activating FoxO3a to activate hepatocellular pyroptosis, and further influence the expression of Caspase1, IL-1β and IL-18.

Key words: reperfusion injury, liver, microRNAs, forkhead transcription factors, miR-182-5p, pyroptosis, FoxO3a