RIG-I,a novel DAMPs sensor for myoglobin,activates NF-κB/caspase-3 signaling in CS-AKI model  

作　　者：Peng-Tao Wang Ning Li Xin-Yue Wang Jia-Le Chen Chen-Hao Geng Zi-Quan Liu Hao-Jun Fan Qi Lv Shi-Ke Hou Yan-Hua Gong 

机构地区：[1]General Hospital of Tianjin Medical University,Tianjin 300052,China [2]institute of Disaster Medicine,Tianjin University,Tianjin 300072,China [3]State Key Laboratory of Medicinal Chemical Biology,Nankai University,Tianjin 300350,China [4]Tianjin Key Laboratory of Disaster Medicine Technology,Tianjin 300072,China [5]Wenzhou Safety(Emergency)Institute,Tianjin University,Wenzhou 325000,Zhejiang China.

出　　处：《Military Medical Research》2022年第1期40-52,共13页军事医学研究（英文版）

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)。

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(P<0.05).However,the levels of interferon regulatory factor 3(IR

关 键 词：Crush syndrome Acute kidney injury Retinoic acid-inducible gene I MYOGLOBIN Nuclear factor kappa-B/caspase-3 Damage-associated molecular patterns 

分 类 号：R642[医药卫生—外科学] R692[医药卫生—临床医学]