| Objective: To investigate the protective effect of emodin on the kidney of rats with early renal ischemia-reperfusion injury(RIRI)and its possible mechanism,emodin was used to interfere with the early RIRI model rats.Methods: 90 male Wistar rats were randomly divided into blank group(Control group),sham operation group(Sham group),model group(RIRI group),emodin group(Emodin group),solvent control group(DMSO group)and inhibitor group(TAK-242 group).Sham group and RIRI group received intraperitoneal injection of 0.8-1ml saline at 1 hour before operation and immediately after operation,while Emodin group,DMSO group and TAK-242 group received intraperitoneal injection of Emodin,DMSO and TAK-242 solution 1 hour before operation and immediately after operation.12 hours later,blood was collected from abdominal aorta and bilateral kidney tissues were removed.Observation of renal histomorphology by HE staining.The expressions of HMGB1,TLR4 and p-IKK β in kidney tissue were detected by immunohistochemical method.The expression levels of HMGB1,TLR4,IKK β,p-IKK β,NF-κB p65 and p-NF-κB p65 in kidney tissue were detected by Western blot,renal cell apoptosis was detected by TUNEL method,and the expression levels of Il-35 and Tnf-α in rat kidney tissue were detected by q RT-PCR method.The contents of IL-35 and TNF-α in serum were detected by ELISA method,and the levels of Cr and BUN in serum were detected by biochemical analyzer.Results:1.HE staining results showed that in the Control group and the Sham group,there was no obvious pathological change in the renal tissue morphology,the glomerular structure was clear,the lumen was not deformed,and the nucleus and cytoplasm had no abnormal changes;in the RIRI group,a large number of dilated renal tubules and epithelial cell necrosis were seen.Shedding,renal interstitium and glomerulus congestion and edema were obvious,brush border structure disappeared,inflammatory cell infiltration and nucleus shedding;in Emodin group,renal interstitial blood stasis and edema were alleviated,brush border structure disappeared,and there was a small amount of stasis In the blood,no inflammatory cell infiltration was seen;in the DMSO group,a large number of necrotic renal tubular cells showed hyalinization,the glomeruli were detached and separated,and the hyperemia and edema were obvious,accompanied by inflammatory cell infiltration;in the TAK-242 group,renal interstitial blood stasis and the degree of edema decreased,the brush border structure disappeared,the detachment and necrosis of renal tubules and glomeruli were alleviated,and there was slight inflammatory cell infiltration.2.The results of immunohistochemistry showed that compared with the Control group,the expression levels of HMGB1,TLR4 and p-IKK β in the RIRI group were significantly increased(P<0.05);compared with the RIRI group,the expression levels of TLR4 and p-IKK β in the Emodin group and the TAK-242 group were significantly lower(P<0.05);compared with the RIRI group,the expression level of HMGB1 in the Emodin group was significantly lower(P<0.05),there was no significant difference in the expression level of HMGB1 in the TAK-242 group(P>0.05).3.The results of Western blot showed that compared with the Control group,there was no significant change in the protein levels of IKKβ and NF-κB p65 in the RIRI group.The expression levels of four proteins,HMGB1,TLR4,p-IKK β,and p-NF-κB p65 were significantly increased(P<0.05).Compared with RIRI group,the level of HMGB1 in Emodin group was significantly decreased(P<0.05),the protein expression levels of TLR4,p-IKK β,p-NF-κB p65 were significantly decreased in Emodin group and TAK-242 group(P<0.05).There was no significant difference in HMGB1 protein level in TAK-242 groups(P>0.05).4.The results of TUNEL experiment showed that compared with the Control group,the level of apoptosis in the RIRI group was significantly increased(P<0.05);Compared with RIRI group,the apoptosis level of Emodin group and TAK-242 group was significantly decreased(P<0.05).5.The results of renal tissue q RT-PCR detection showed that compared with the Control group,the levels of Il-35 and Tnf-αm RNA in the RIRI group were significantly increased(P<0.05);Compared with RIRI group,the m RNA levels of Il-35 and Tnf-α m RNA in Emodin group and TAK-242 group were significantly decreased(P<0.05).6.The serum ELISA test results showed that compared with the Control group,the level of IL-35 in the RIRI group was significantly lower,and the TNF-α level was significantly increased(P<0.05);compared with the RIRI group,the IL-35 levels in the Emodin group and the TAK-242 group were significantly increased,and the TNF-α levels were significantly decreased(P<0.05).7.The detection results of renal function indicators showed that compared with the Control group,the levels of Cr and BUN in the RIRI group were significantly increased(P<0.05);Compared with RIRI group,the levels of Cr and BUN in Emodin group and TAK-242 group were significantly lower(P<0.05).Conclusion: Emodin has a good protective effect on the renal injury of rats in the early stage of RIRI,and the mechanism may be related to the regulation of the expression of HMGB1/TLR4/NF-κB inflammatory pathway by emodin. |
PDF Full Text Request