RIG-I,a novel DAMPs sensor for myoglobin,activates NF-κB/caspase-3 signaling in CS-AKI model
RIG-I,a novel DAMPs sensor for myoglobin, activates NF-κB/caspase-3 signaling in CS-AKI model作者机构：General Hospital of Tianjin Medical UniversityTianjin 300052China institute of Disaster MedicineTianjin UniversityTianjin 300072China State Key Laboratory of Medicinal Chemical BiologyNankai UniversityTianjin 300350China Tianjin Key Laboratory of Disaster Medicine TechnologyTianjin 300072China Wenzhou Safety(Emergency)InstituteTianjin UniversityWenzhou 325000Zhejiang China.
出 版 物：《Military Medical Research》 (军事医学研究（英文版）)
年 卷 期：2022年第9卷第1期
页 面：40-52页
核心收录：
基 金：supported by the Tianjin UniversityaDouble First Classoconstruction talent start-up fund to Dr.Yan-Hua Gong,the grants awarded to Shi-Ke Hou by Science and Technology Commission of the CMC(c12019048) Ning Li by Open Fund of State Key Laboratory of Medicinal Chemical Biology(Nankai University)(2020010)。
主 题：Crush syndrome Acute kidney injury Retinoic acid-inducible gene I Myoglobin Nuclear factor kappa-B/caspase-3 Damage-associated molecular patterns
摘 要：Background:Acute kidney injury(AKI)is the main life-threatening complication of crush syndrome(CS),and myoglobin is accepted as the main pathogenic factor.The pattern recognition receptor retinoicacid-inducible gene I(RIG-I)has been reported to exert anti-viral effects function in the innate immune response.However,it is not clear whether RIG-I plays a role in CS-AKI.The present research was carried out to explore the role of RIG-I in CS-AKI.Methods:Sprague-Dawley rats were randomly divided into two groups:the sham and CS groups(n=12).After administration of anesthesia,the double hind limbs of rats in the CS group were put under a pressure of 3 kg for 16 h to mimic crush conditions.The rats in both groups were denied access to food and water.Rats were sacrificed at 12 h or 36 h after pressure was relieved.The successful establishment of the CS-AKI model was confirmed by serum biochemical analysis and renal histological examination.In addition,RNA sequencing was performed on rat kidney tissue to identify molecular pathways involved in CS-AKI.Furthermore,NRK-52 E cells were treated with 200μmol/L ferrous myoglobin to mimic CS-AKI at the cellular level.The cells and cell supernatant samples were collected at 6 h or 24 h.Small interfering RNAs(siRNA)was used to knock down RIG-I expression.The relative expression levels of molecules involved in the RIG-I pathway in rat kidney or cells samples were measured by quantitative real-time PCR(qPCR),Western blotting analysis,and immunohistochemistry(IHC)staining.Tumor necrosis factor-α(TNF-α)was d etected by ELISA.Co-immunoprecipitation(Co-IP)assays were used to detect the interaction between RIG-I and myoglobin.Results:RNA sequencing of CS-AKI rat kidney tissue revealed that the different expression of RIG-I signaling pathway.qPCR,Western blotting,and IHC assays showed that RIG-I,nuclear factor kappa-B(NF-κB)P65,p-P65,and the a poptotic marker caspase-3 and cleaved caspase-3 were up-regulated in the CS group(P0.05).However,the levels of interferon regulatory factor 3(IRF3),p-IRF3 and the antiviral factor interferon-beta(IFN-β)showed no significant c hanges between the sham and CS groups.Co-IP assays showed the interaction between RIG-I and myoglobin in the kidneys of the CS group.Depletion of RIG-I could alleviate the myoglobin induced expression of apoptosis-associated molecules via the NF-κB/caspase-3 axis.C onclusions:RIG-I is a novel damage-associated molecular patterns(DAMPs)sensor for myoglobin and participates in the NF-κB/caspase-3 signaling pathway in CS-AKI.In the development of CS-AKI,specific intervention in the RIG-I p athway might be a potential therapeutic strategy for CS-AKI.
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