PMID- 38347637 OWN - NLM STAT- MEDLINE DCOM- 20240220 LR - 20240320 IS - 1478-811X (Electronic) IS - 1478-811X (Linking) VI - 22 IP - 1 DP - 2024 Feb 12 TI - Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury. PG - 121 LID - 10.1186/s12964-023-01443-3 [doi] LID - 121 AB - OBJECTIVES: To explore whether the gap junction (GJ) composed by connexin32(Cx32) mediated pyroptosis in renal ischemia-reperfusion(I/R) injury via transmitting miR155-3p, with aim to provide new strategies for the prevention and treatment of acute kidney injury (AKI) after renal I/R. METHODS: 8-10 weeks of male C57BL/ 6 wild-type mice and Cx32 knockdown mice were divided into two groups respectively: control group and renal I/R group. MCC950 (50 mg/kg. ip.) was used to inhibit NLRP3 in vivo. Human kidney tubular epithelial cells (HK - 2) and rat kidney tubular epithelial cells (NRK-52E) were divided into high-density group and low-density group, and treated with hypoxia reoxygenation (H/R) to mimic I/R. The siRNA and plasmid of Cx32, mimic and inhibitor of miR155-3p were transfected into HK - 2 cells respectively. Kidney pathological and functional injuries were measured. Western Blot and immunofluorescent staining were used to observe the expression of NLRP3, GSDMD, GSDMD-N, IL - 18, and mature IL-18. The secretion of IL-18 and IL-1beta in serum, kidney tissue and cells supernatant were detected by enzyme-linked immuno sorbent assay (ELISA) kit, and the expression of NLPR3 and miR155-3p were detected by RT-qPCR and fluorescence in situ hybridization (FISH). RESULTS: Tubular pyroptosis were found to promote AKI after I/R in vivo and Cx32-GJ regulated pyroptosis by affecting the expression of miR155-3p after renal I/R injury. In vitro, H/R could lead to pyroptosis in HK-2 and NRK-52E cells. When the GJ channels were not formed, and Cx32 was inhibited or knockdown, the expression of miR155-3p was significantly reduced and the pyroptosis was obviously inhibited, leading to the reduction of injury and the increase of survival rate. Moreover, regulating the level of miR155-3p could affect survival rate and pyroptosis in vitro after H/R. CONCLUSIONS: The GJ channels composed of Cx32 regulated tubular pyroptosis in renal I/R injury by transmitting miR155-3p. Inhibition of Cx32 could reduce the level of miR155-3p further to inhibit pyroptosis, leading to alleviation of renal I/R injury which provided a new strategy for preventing the occurrence of AKI. Video Abstract. CI - (c) 2024. The Author(s). FAU - Chen, Liubing AU - Chen L AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Fang, Hongyi AU - Fang H AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Li, Xiaoyun AU - Li X AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Yu, Peiling AU - Yu P AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Guan, Yu AU - Guan Y AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Xiao, Cuicui AU - Xiao C AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Deng, Zhizhao AU - Deng Z AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Hei, Ziqing AU - Hei Z AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. FAU - Chen, Chaojin AU - Chen C AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. chenchj28@mail.sysu.edu.cn. FAU - Luo, Chenfang AU - Luo C AD - Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen University, No. 600 Tianhe Road, Guangzhou, 510630, Guangdong Province, China. luochenf@mail.sysu.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Video-Audio Media DEP - 20240212 PL - England TA - Cell Commun Signal JT - Cell communication and signaling : CCS JID - 101170464 RN - 0 (Interleukin-18) RN - 0 (MicroRNAs) RN - 0 (MIRN155 microRNA, human) RN - 0 (Mirn155 microRNA, mouse) RN - 0 (MIRN155 microRNA, rat) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (GJB1 protein, human) RN - 0 (Gjb1 protein, mouse) RN - 0 (Gjb1 protein, rat) SB - IM MH - Animals MH - Humans MH - Male MH - Mice MH - Rats MH - *Acute Kidney Injury/genetics MH - Gap Junctions/metabolism MH - Hypoxia MH - In Situ Hybridization, Fluorescence MH - Interleukin-18/genetics MH - Kidney/metabolism MH - Mice, Inbred C57BL MH - *MicroRNAs/genetics/metabolism MH - NLR Family, Pyrin Domain-Containing 3 Protein/metabolism MH - Pyroptosis MH - *Reperfusion Injury/metabolism PMC - PMC10863161 OTO - NOTNLM OT - Connexin32 OT - Hypoxia and reoxygenation injury OT - Pyroptosis OT - Renal ischemia reperfusion injury OT - miR155-3p COIS- The authors declare no competing interests. EDAT- 2024/02/13 00:42 MHDA- 2024/02/13 06:45 PMCR- 2024/02/12 CRDT- 2024/02/12 23:49 PHST- 2023/06/07 00:00 [received] PHST- 2023/12/13 00:00 [accepted] PHST- 2024/02/13 06:45 [medline] PHST- 2024/02/13 00:42 [pubmed] PHST- 2024/02/12 23:49 [entrez] PHST- 2024/02/12 00:00 [pmc-release] AID - 10.1186/s12964-023-01443-3 [pii] AID - 1443 [pii] AID - 10.1186/s12964-023-01443-3 [doi] PST - epublish SO - Cell Commun Signal. 2024 Feb 12;22(1):121. doi: 10.1186/s12964-023-01443-3.