PMID- 34148549 OWN - NLM STAT- MEDLINE DCOM- 20211027 LR - 20211027 IS - 2054-9369 (Electronic) IS - 2095-7467 (Print) IS - 2054-9369 (Linking) VI - 8 IP - 1 DP - 2021 Jun 21 TI - RIG-I, a novel DAMPs sensor for myoglobin activates NF-kappaB/caspase-3 signaling in CS-AKI model. PG - 37 LID - 10.1186/s40779-021-00333-4 [doi] LID - 37 AB - 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-52E cells were treated with 200 mumol/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-alpha (TNF-alpha) was detected 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-kappaB) P65, p-P65, and the apoptotic 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 (IRF3), p-IRF3 and the antiviral factor interferon-beta (IFN-beta) showed no significant changes 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-kappaB/caspase-3 axis. CONCLUSION: RIG-I is a novel damage-associated molecular patterns (DAMPs) sensor for myoglobin and participates in the NF-kappaB/caspase-3 signaling pathway in CS-AKI. In the development of CS-AKI, specific intervention in the RIG-I pathway might be a potential therapeutic strategy for CS-AKI. FAU - Wang, Peng-Tao AU - Wang PT AD - General Hospital of Tianjin Medical University, Tianjin, 300052, China. FAU - Li, Ning AU - Li N AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300350, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. FAU - Wang, Xin-Yue AU - Wang XY AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. FAU - Chen, Jia-Le AU - Chen JL AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. FAU - Geng, Chen-Hao AU - Geng CH AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. FAU - Liu, Zi-Quan AU - Liu ZQ AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. FAU - Fan, Hao-Jun AU - Fan HJ AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. FAU - Lv, Qi AU - Lv Q AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. FAU - Hou, Shi-Ke AU - Hou SK AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. houshike@tju.edu.cn. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. houshike@tju.edu.cn. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. houshike@tju.edu.cn. FAU - Gong, Yan-Hua AU - Gong YH AD - Institute of Disaster Medicine, Tianjin University, Tianjin, 300072, China. gongyanhua@tju.edu.cn. AD - Tianjin Key Laboratory of Disaster Medicine Technology, Tianjin, 300072, China. gongyanhua@tju.edu.cn. AD - Wenzhou Safety (Emergency) Institute, Tianjin University, Wenzhou, 325000, China. gongyanhua@tju.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210621 PL - England TA - Mil Med Res JT - Military Medical Research JID - 101643181 RN - 0 (Alarmins) RN - 0 (Myoglobin) RN - 0 (NF-kappa B) RN - EC 2.7.7.- (RIG-I protein, rat) RN - EC 3.4.22.- (Casp3 protein, rat) RN - EC 3.4.22.- (Caspase 3) RN - EC 3.6.4.13 (RNA Helicases) SB - IM MH - Acute Kidney Injury/etiology/physiopathology MH - Alarmins MH - Animals MH - Caspase 3/*drug effects MH - China MH - Crush Syndrome/blood/complications MH - Disease Models, Animal MH - Male MH - Myoglobin/pharmacology/therapeutic use MH - NF-kappa B/*drug effects MH - RNA Helicases/*pharmacology/therapeutic use MH - Rats MH - Rats, Sprague-Dawley MH - Signal Transduction/*drug effects PMC - PMC8215750 OTO - NOTNLM OT - Acute kidney injury OT - Crush syndrome OT - Damage-associated molecular patterns OT - Myoglobin OT - Nuclear factor kappa-B/caspase-3 OT - Retinoic acid-inducible gene I COIS- The authors declare that there are no competing interests. EDAT- 2021/06/22 06:00 MHDA- 2021/10/28 06:00 PMCR- 2021/06/21 CRDT- 2021/06/21 05:27 PHST- 2020/12/25 00:00 [received] PHST- 2021/06/10 00:00 [accepted] PHST- 2021/06/21 05:27 [entrez] PHST- 2021/06/22 06:00 [pubmed] PHST- 2021/10/28 06:00 [medline] PHST- 2021/06/21 00:00 [pmc-release] AID - 10.1186/s40779-021-00333-4 [pii] AID - 333 [pii] AID - 10.1186/s40779-021-00333-4 [doi] PST - epublish SO - Mil Med Res. 2021 Jun 21;8(1):37. doi: 10.1186/s40779-021-00333-4.