| Objective:Intestinal ischaemia-reperfusion injury is a common and high mortality disease in surgery as well as in critical care units,and its occurrence and development are closely related to the inflammatory response and apoptosis.In recent years,a new gastrointestinal hormone Ghrelin,the growth hormone-secreting receptor,has been identified as a biodiverse agent that protects the intestine in terms of improving the inflammatory response and regulating cell proliferation.We investigated whether this growth hormone-secreting receptor could reduce intestinal ischemia-reperfusion injury through the inhibition of TLR4/NLRP3 signalling pathway.Methods:We established a rat model of intestinal ischemia/reperfusion.There are 18 SD male rats randomly divided into three groups,a sham group,an I/R group and a Ghrelin intervention group.The intestinal I/R animal model was confirmed by clamping the superior mesenteric artery for 60 minutes,followed by reperfusion for 2hours.While Ghrelin was injected intraperitoneally for intervention.The morphological changes of the small intestine were observed by hematoxylin eosin(HE)staining and the extent of intestinal mucosal damage was evaluated according to Chiu’s method.expression levels.TUNEL assay was used to detect apoptosis in intestinal cells.Results1.The results of HE staining showed that the intestinal mucosal injury was significantly aggravated and the intestinal mucosal Chiu’s score was significantly higher in the I/R group compared to the Sham group;the intestinal mucosal injury was significantly reduced and the intestinal mucosal Chiu’s score was significantly lower in the I/R+G group compared to the I/R group,all with statistically significant differences(P < 0.05).2.TUNEL staining results showed that apoptosis was less in the Sham group,with an apoptosis rate of 5.90±7.92(%);apoptosis was significantly more in the I/R3.group,with an apoptosis rate of 67.60 ± 10.00(%);and apoptosis was substantially reduced in the I/R+G group,with an apoptosis rate of 16.80±3.96(%).All differences were statistically significant(P < 0.05).4.The ELISA results showed that the levels of pro-inflammatory cytokines IL-6、TNF-αand IL-1 βin serum were remarkably higher in the I/R and I/R+G groups compared with the Sham group,and the difference was found to be statistically significant(P < 0.05);compared with the I/R group,results showed that serum levels of IL-6、TNF-αand IL-1βwere significantly lower in the I / R + G group,and the difference was found to be statistically significant(P < 0.05).5.Western Blot assay results: the relative protein content of Caspase-1,TLR4,and NLRP3 increased in the I/R group;Ghrelin significantly inhibited the protein expression of TLP4,NLRP3 and caspase-1,and the difference was statistically significant(P < 0.05);compared with the Sham group,the relative protein contents of caspase-1,TLP4 and NLRP3 were increased in the I/R group-1,TLP4 and NLRP3 were significantly increased in the I/R group compared with the Sham group,and the relative amounts of all three proteins were up-regulated in the I/R+G group,and the discrepancies were all of statistical importance(P < 0.05).Conclusion:Ghrelin attenuates histopathological changes in the intestinal mucosa of rats with intestinal ischemia / reperfusion;Ghrelin decreased the expression of inflammatory factors in the serum of rats with intestinal ischemia/reperfusion and may reduce intestinal ischemia/reperfusion injury by inhibiting inflammatory response and apoptosis through TLR4/NLRP3 signaling pathway,which may provide new therapeutic targets and strategies for the treatment of intestinal I/R injury. |
