Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury
Abstract 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. Method...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-02-01
|
| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12964-023-01443-3 |
| _version_ | 1797273851854323712 |
|---|---|
| author | Liubing Chen Hongyi Fang Xiaoyun Li Peiling Yu Yu Guan Cuicui Xiao Zhizhao Deng Ziqing Hei Chaojin Chen Chenfang Luo |
| author_facet | Liubing Chen Hongyi Fang Xiaoyun Li Peiling Yu Yu Guan Cuicui Xiao Zhizhao Deng Ziqing Hei Chaojin Chen Chenfang Luo |
| author_sort | Liubing Chen |
| collection | DOAJ |
| description | Abstract 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-1β 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 |
| first_indexed | 2024-03-07T14:50:04Z |
| format | Article |
| id | doaj.art-ed0234641f7b4ee69a8230b3bc946aeb |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-03-07T14:50:04Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-ed0234641f7b4ee69a8230b3bc946aeb2024-03-05T19:46:34ZengBMCCell Communication and Signaling1478-811X2024-02-0122111610.1186/s12964-023-01443-3Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injuryLiubing Chen0Hongyi Fang1Xiaoyun Li2Peiling Yu3Yu Guan4Cuicui Xiao5Zhizhao Deng6Ziqing Hei7Chaojin Chen8Chenfang Luo9Department of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityDepartment of Anesthesiology, The Third Affiliated Hospital of Sun Yat-sen UniversityAbstract 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-1β 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 Abstracthttps://doi.org/10.1186/s12964-023-01443-3Renal ischemia reperfusion injuryHypoxia and reoxygenation injuryConnexin32PyroptosismiR155-3p |
| spellingShingle | Liubing Chen Hongyi Fang Xiaoyun Li Peiling Yu Yu Guan Cuicui Xiao Zhizhao Deng Ziqing Hei Chaojin Chen Chenfang Luo Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury Cell Communication and Signaling Renal ischemia reperfusion injury Hypoxia and reoxygenation injury Connexin32 Pyroptosis miR155-3p |
| title | Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury |
| title_full | Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury |
| title_fullStr | Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury |
| title_full_unstemmed | Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury |
| title_short | Connexin32 gap junction channels deliver miR155-3p to mediate pyroptosis in renal ischemia-reperfusion injury |
| title_sort | connexin32 gap junction channels deliver mir155 3p to mediate pyroptosis in renal ischemia reperfusion injury |
| topic | Renal ischemia reperfusion injury Hypoxia and reoxygenation injury Connexin32 Pyroptosis miR155-3p |
| url | https://doi.org/10.1186/s12964-023-01443-3 |
| work_keys_str_mv | AT liubingchen connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT hongyifang connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT xiaoyunli connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT peilingyu connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT yuguan connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT cuicuixiao connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT zhizhaodeng connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT ziqinghei connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT chaojinchen connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury AT chenfangluo connexin32gapjunctionchannelsdelivermir1553ptomediatepyroptosisinrenalischemiareperfusioninjury |