Objective To investigate the role of BAM15,a novel mitochondrial uncoupler,in is-chemia-reperfusion injury induced acute kidney injury.Methods Totally,28 C57 mice(6-8 weeks old,weight 24-26 g)were randomly divided into normal control(NC)group,ischemia-reperfusion group,BAM15-treated group,and ischemia-reperfusion+BAM15 treated group,with saline injected intraperito-neally in the NC group.The clamp was removed after 60 min ligation of both renal arteries and veins in the ischemia-reperfusion group.BAM15(5 mg/kg)was injected intraperitoneally in the BAM15-treated group.BAM15(5 mg/kg)was given immediately after the ischemia-reperfusion model was established in the is-chemia-reperfusion+BAM15 treatment group.The 5-day survival rates of mice in different groups were compared.Creatinine(Cr)and blood urea nitrogen(BUN)levels were detected by biochemical method.Hematoxylin-eosin(HE)staining was used to evaluate renal tubule injury,and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)staining was used to evaluate renal cell apoptosis.Tissue levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6 and IL-18 were determined by en-zyme-linked immunosorbent assay.The expression levels of cyclic GMP-AMP synthase(cGAS)and inter-feron stimulating factor(STING)were detected by immunohistochemical staining.One-way ANOVA was used for comparison among groups,and Log-rank(Mantel-Cox)test was used for survival analysis.Results The 5-day survival rate was 1/8 in the ischemia-reperfusion group and 5/8 in the ischemia-reperfusion+BAM15 group,and the difference was statistically significant(P＜0.05).HE staining showed that renal tubule injury in the ischemia-reperfusion group was significantly severer than in the control group(2.56± 0.34 vs.0±0,t=4.154,P＜0.05)and also severer than in the ischemia-reperfusion+BAM15 group(2.56±0.34 vs.1.65±0.21,t=3.217,P＜0.05).TUNEL staining showed that apoptosis rate in the ischemia-reperfusion group was significantly higher than in the control group(8.23±1.14 vs.0±0,t=10.068,P＜0.05)and also higher than in the ischemia-reperfusion+BAM15 group(8.23±1.14 vs.4.68±0.61,t=6.144,P＜0.05).ELISA results showed that the levels of TNF-α,IL-6,IL-1β and IL-18 in the ischemia-reperfusion+BAM15 treatment group were significantly decreased as compared with those in the ischemia-reperfusion group(P＜0.05).Immunohistochemical staining results showed that the cGAS positive expression area in the ischemia-reperfusion group was significantly larger than that in the control group(13.23±1.87 vs.3.72±0.39,t=11.146,P＜0.05),and STING positive expression are-a was significantly larger than that in the control group(15.11±1.98 vs.3.11±0.41,t=14.214,P＜0.05),and the difference was statistically significant.The cGAS positive expression area in the ischemia-reperfusion group was larger than that in the ischemia-reperfusion+BAM15 group[(13.23±1.87)％vs.(8.31±1.12)％,t=8.625,P＜0.05],and STING positive expression area in the ischemia-reperfusion+BAM15 group was larger than that in the ischemia-reperfusion+BAM15 group[(15.11±1.98)％vs.(9.21±1.02)％,t=8.541,P＜0.05],the difference was statistically significant.Conclusion BAM15 may alleviate ischemia-reperfusion injury induced acute kidney injury by inhibiting inflammation and apoptosis.
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