The Protective Effect Of NLRP3 Inhibitor CY-09 On Liver Injury After Renal Ischemia Reperfusion Injury In Mice

Objective: To investigate the mechanism of liver injury after renal ischemia reperfusion(R/IR)injury in mice and the positive effect of CY-09 which is the specific inhibitor of NOD-like receptor thermal protein domain associated protein 3(NLRP3).Methods: Thirty two Bal/C mice were were randomly divided into four groups:Sham group,Sham with NLRP3 inhibitor group(Sham+CY-09),R/IR group,R/IR with NLRP3 inhibitor group(R/IR+CY-09).Each group contains eight mice.Sham +CY-09 group and R/IR + CY-09 group were injected CY-09(50ug/kg)intraperitoneally at 30 min before surgery.Sham group and R/IR group were injected with saline with equal volume.The right kidney was resected in all groups,the left kidney was fully exposed and clipped for45 minutes in the R/IR group and R/IR +CY-09 group.The left renal ischemia for 45 minutes and reperfuse for 24 hours.Serum creatinine(Scr) and Alaninetransaminase(ALT)concentration were detected by Kits.Liver and kidney histopathological injury were detected by HE staining.Apoptosis of liver tissue were obeserved by Tunel stain.The proteins level of NLRP3,ASC and Caspase-1 was determined by Western blot.The content of IL-1β and IL-6 inflammatory factors were detected by ELISA kits.Results: Compared with the Sham group,the levels of Scr and ALT in the R/IR group were significantly increased(P<0.0001),HE staining showed obvious pathological damage of liver and kidney tissues,Tunel staining showed a significant increase in liver tissue apoptosis(P<0.0001),In liver tissue,the protein expressions of NLRP3(P<0.01),Caspase-1(P<0.05),and ASC(P<0.05)were significantly increased,and the inflammatory factors IL-1β(P<0.01),IL-6(P<0.001)levels were significantly increased;the expression levels of miR687-3p(P<0.01),miR24-3p(P<0.05),and miR126-3p(P<0.01)in liver and kidney tissues were significantly increased,while the expression levels of miR494-3p in liver tissue decreased significantly(P<0.05),and there was no significant difference in the expression level of miR494-3p in kidney tissue(P>0.05).Compared with the R/IR group,the levels of Scr and ALT in the R/IR +CY-09 group decreased significantly(P <0.01),HE staining showed a reduction in pathological damage to liver and kidney tissue,Tunel staining showed a decrease in liver tissue apoptosis,NLRP3(P <0.01),Caspase-1(P <0.05),ASC(P <0.05) protein content in liver tissue decreased significantly.The content of inflammatory factor IL-1β decreased significantly(P <0.05),while the level of the inflammatory factor IL-6 showed no difference(P>0.05).Conclusion(s): Renal ischemia reperfusion can activate the NLRP3 inflammasome in liver tissue to promote inflammation-induced liver damage,and pretreatment with NLRP3 inhibitor CY-09 can alleviate liver tissue damage.