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RiG-i,a novel DAMPs sensor for myoglobin,activates NF-κB/caspase-3 signaling in CS-AKi model

Military Medical Research 2022年第1期9卷 40-52页

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 general Hospital of Tianjin Medical University Tianjin 300052China institute of Disaster Medicine Tianjin UniversityTianjin 300072China State Key Laboratory of Medicinal Chemical Biology Nankai UniversityTianjin 300350China Tianjin Key Laboratory of Disaster Medicine Technology Tianjin 300072China Wenzhou Safety(Emergency)institute Tianjin UniversityWenzhou 325000Zhejiang China.

Background:Acute kidney injury(AKi)is the main life-threatening complication of crush syndrome(CS),and myoglobin is accepted as the main pathogenic *** pattern recognition receptor retinoicacid-inducible gene i(RiG-i)has been reported to exert anti-viral effects function in the innate immune ***,it is not clear whether RiG-i plays a role in *** present research was carried out to explore the role of RiG-i in ***: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 *** rats in both groups were denied access to food and *** were sacrificed at 12 h or 36 h after pressure was *** successful establishment of the CS-AKi model was confirmed by serum biochemical analysis and renal histological *** addition,RNA sequencing was performed on rat kidney tissue to identify molecular pathways involved in ***,NRK-52 E cells were treated with 200μmol/L ferrous myoglobin to mimic CS-AKi at the cellular *** cells and cell supernatant samples were collected at 6 h or 24 *** interfering RNAs(siRNA)was used to knock down RiG-i *** 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)*** necrosis factor-α(TNF-α)was d etected by ***-immunoprecipitation(Co-iP)assays were used to detect the interaction between RiG-i and ***:RNA sequencing of CS-AKi rat kidney tissue revealed that the different expression of RiG-i signaling ***,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

关键词： Crush syndrome Acute kidney injury retinoic acid-inducible gene i Myoglobin Nuclear factor kappa-B/caspase-3 Damage-associated molecular patterns

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(-)-Epigallocatechin-3-gallate enhances poly i:C-induced interferon-λ1 production and inhibits hepatitis C virus replication in hepatocytes

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World Journal of Gastroenterology 2017年第32期23卷 5895-5903页

作者： Yi-Zhong Wang Jie-Liang Li Xu Wang Ting Zhang Wen-Zhe Ho Department of infectious Diseases Shanghai Children’s Hospital Shanghai Jiao Tong University Department of Pathology and Laboratory Medicine Temple University Lewis Katz School of Medicine

AiM To investigate the effect of(-)-epigallocatechin-3-gallate(EGCG) on polyinosinic-polycytidylic acid(poly i:C)-triggered intracellular innate immunity against hepatitis C virus(HCV) in hepatocytes. METHODS A cell culture model of HCV infection was generated by infecting a hepatoma cell line, Huh7, with HCV JFH-1 strain(JFH-1-Huh7). Poly i:C with a high molecular weight and EGCG were used to stimulate the JFH-1-Huh7 cells. Real-time reverse transcription-polymerase chain reaction was used to detect the expression levels of intracellular m RNAs and of intracellular and extracellular HCV RNA. Enzyme-linked immunosorbent assay was used to evaluate the interferon(iFN)-λ1 protein level in the cell culture supernatant. immunostaining was used to examine HCV core protein expression in Huh7 *** Our recent study showed that HCV replication could impair poly i:C-triggered intracellular innate immune responses in hepatocytes. in the current study, we showed that EGCG treatment significantly increased the poly i:C-induced expression of Toll-like receptor 3(TLR3), retinoic acid-inducible gene i, and iFN-λ1 in JFH-1-Huh7 cells. in addition, supplementation with EGCG increased the poly i:C-mediated antiviral activity in JFH-1-Huh7 cells at the intracellular and extracellular HCV RNA and protein levels. Further investigation of the mechanisms showed that EGCG treatment significantly enhanced the poly i:C-induced expression of iFN-regulatory factor 9 and several antiviral iFNstimulated genes, including iSG15, iSG56, myxovirus resistance A, and 2'-5'-oligoadenylate synthetase 1, which encode the key antiviral elements in the iFN signaling pathway. CONCLUSiON Our observations provide experimental evidence that EGCG has the ability to enhance poly i:C-induced intracellular antiviral innate immunity against HCV replication in hepatocytes.

关键词： (-)-Epigallocatechin-3-gallate Toll-like receptor 3 retinoic acid-inducible gene i iFN-λ1 Hepatitis C virus iFN-stimulated genes

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