Establish An Animal Model Of Contrast-associated Acute Kidney Injury Following Acute Myocardial Infarction And Explore The Role Of Pyroptosis

BackgroundContrast-associated acute kidney injury(CA-AKI)following acute myocardial infarction(AMI),characterized by marked oxidative stress and inflammation disease,is related to adverse outcomes.Pyroptosis is characterized as inflammasome-dependent programmed cell death and mediated by gasdermin D(GSDMD)protein that can occur in AKI.However,the relationship between the pathogenesis of CA-AKI after AMI and pyroptosis is still unclear.Most of the previously established animal models primarily focused on one single factor that causes kidney damage,there is no reliable and stable animal model of CA-AKI after AMI at present.This study aims to establish an animal model of CA-AKI after AMI by combining IRI(ischemia reperfusion injury)surgery with CM(contrast medium)and exploring the role of pyroptosis as well as construct the ceRNA network related to pyroptosis.MethodEighty rats were randomly divided into 4 groups:SHAM group(sham operation+normal saline,n=20),CM group(sham operation+CM,n=20),IRI group(IRI surgery+normal saline,n=20)and CM-IRI group(IRI surgery+CM,n=20).TTC/Evans blue staining and HE staining were used to detecting myocardial and kidney injury,respectively.Western Blot(WB)was used to compare indicators of kidney injury KIM-1 and NGAL.Renal pyroptosis was observed by transmission electron microscope and the expression of pyroptosis protein GSDMD was detected by immunohistochemistry(IHC).Subsequently,enrolled the patients of CA-AKI after AMI and the control patients.Microarray was used to detect and analysis the differentially expressed mRNAs and miRNAs.Pyroptosis association gene datasets were screened based on MSigDB database,then pyroptosis-related differentially expressed genes(py-DEGs)were obtained by intersection with DEmRN As.Pyroptosisrelated DEmRNAs(py-DEmRNAs)and DEmiRNAs(py-DEmiRNAs)were screened by Pearson correlation analysis,and functional enrichment analysis of these key genes was perfonned.Based on the gene interactions,a ceRNA network related to pyroptosis was constructed.ResultTTC/Evans blue double staining results showed the ischemic area and infarct area in the CM-IRI group,and IRI group,while the normal myocardial tissue was shown in the SHAM group and CM group.WB results showed that the expression levels of NGAL and KIM-1 were significantly increased in the CM-IRI group compared with other groups(p<0.001).Transmission electron microscope observed the morphological changes in the kidney of CA-AKI after AMI,such as cell swelling,cell membrane bleb and production of protrusions,and the number of pyroptosis bodies was more in the CM-IRI group.The expression level of GSDMD by IHC was significantly higher in the CM-IRI group than in the control group(p<0.001).83 DEmiRNAs and 5983 DEmRNAs were identified according to the microarray analysis.Pearson correlation analysis showed a total of 10 py-DEmiRNAs and 517 pyDEmRNAs.According to the gene interaction relationships obtained from the database,a total of 15 lncRNAs,61 circRNAs,8 miRNAs and 7 mRNAs were identified to be involved in the construction of pyroptosis-related ceRNA regulatory network.ConclusionIn this study,an animal model close to the clinical situation of CA-AKI after AMI was established by combining contrast media and ischemia-reperfusion surgery and the role of pyroptosis in CA-AKI after AMI was also verified.The pyroptosis-related ceRNA network was constructed,and these results can provide a new basis for molecular biology research on the diagnosis and treatment of CA-AKI after AMI.