CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway
BackgroundAcute kidney injury is a common and severe complication of sepsis. Sepsis -induced acute kidney injury(S-AKI) is an independent risk factor for mortality among sepsis patients. However, the mechanisms of S-AKI are complex and poorly understand. Therefore, exploring the underlying mechanism...
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| Format: | Article |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1053754/full |
| _version_ | 1797954023448379392 |
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| author | Yuting Wang Wenjie Xi Xinyi Zhang Xinwen Bi Boyang Liu Xiaoming Zheng Xinjin Chi |
| author_facet | Yuting Wang Wenjie Xi Xinyi Zhang Xinwen Bi Boyang Liu Xiaoming Zheng Xinjin Chi |
| author_sort | Yuting Wang |
| collection | DOAJ |
| description | BackgroundAcute kidney injury is a common and severe complication of sepsis. Sepsis -induced acute kidney injury(S-AKI) is an independent risk factor for mortality among sepsis patients. However, the mechanisms of S-AKI are complex and poorly understand. Therefore, exploring the underlying mechanisms of S-AKI may lead to the development of therapeutic targets.MethodA model of S-AKI was established in male C57BL/6 mice using cecal ligation and puncture (CLP). The data-independent acquisition (DIA)-mass spectrometry-based proteomics was used to explore the protein expression changes and analyze the key proteomics profile in control and CLP group. The methodology was also used to identify the key proteins and pathways. S-AKI in vitro was established by treating the HK-2 cells with lipopolysaccharide (LPS). Subsequently, the effect and mechanism of Cathepsin B (CTSB) in inducing apoptosis in HK-2 cells were observed and verified.ResultsThe renal injury scores, serum creatinine, blood urea nitrogen, and kidney injury molecule 1 were higher in septic mice than in non-septic mice. The proteomic analysis identified a total of 449 differentially expressed proteins (DEPs). GO and KEGG analysis showed that DEPs were mostly enriched in lysosomal-related cell structures and pathways. CTSB and MAPK were identified as key proteins in S-AKI. Electron microscopy observed enlarged lysosomes, swelled and ruptured mitochondria, and cytoplasmic vacuolization in CLP group. TUNEL staining and CTSB activity test showed that the apoptosis and CTSB activity were higher in CLP group than in control group. In HK-2 cell injury model, the CTSB activity and mRNA expression were increased in LPS-treated cells. Acridine orange staining showed that LPS caused lysosomal membrane permeabilization (LMP). CA074 as an inhibitor of CTSB could effectively inhibit CTSB activity. CCK8 and Annexin V/PI staining results indicated that CA074 reversed LPS-induced apoptosis of HK-2 cells. The JC-1 and western blot results showed that LPS inhibited mitochondrial membrane potential and activated mitochondrial apoptosis pathway, which could be reversed by CA074.ConclusionsLMP and CTSB contribute to pathogenesis of S-AKI. LPS treatment induced HK-2 cell injury by activating mitochondrial apoptosis pathway. Inhibition of CTSB might be a new therapeutic strategy to alleviate sepsis-induced acute kidney injury. |
| first_indexed | 2024-04-10T23:12:21Z |
| format | Article |
| id | doaj.art-7b8068eb415544e0a9576b94c931b691 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-10T23:12:21Z |
| publishDate | 2023-01-01 |
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| series | Frontiers in Immunology |
| spelling | doaj.art-7b8068eb415544e0a9576b94c931b6912023-01-13T05:02:47ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-01-011310.3389/fimmu.2022.10537541053754CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathwayYuting Wang0Wenjie Xi1Xinyi Zhang2Xinwen Bi3Boyang Liu4Xiaoming Zheng5Xinjin Chi6Department of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaDepartment of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaScientific Research Center, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaDepartment of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaDepartment of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaScientific Research Center, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaDepartment of Anesthesiology, The Seventh Affiliated Hospital of Sun Yat-sen University, Shenzhen, ChinaBackgroundAcute kidney injury is a common and severe complication of sepsis. Sepsis -induced acute kidney injury(S-AKI) is an independent risk factor for mortality among sepsis patients. However, the mechanisms of S-AKI are complex and poorly understand. Therefore, exploring the underlying mechanisms of S-AKI may lead to the development of therapeutic targets.MethodA model of S-AKI was established in male C57BL/6 mice using cecal ligation and puncture (CLP). The data-independent acquisition (DIA)-mass spectrometry-based proteomics was used to explore the protein expression changes and analyze the key proteomics profile in control and CLP group. The methodology was also used to identify the key proteins and pathways. S-AKI in vitro was established by treating the HK-2 cells with lipopolysaccharide (LPS). Subsequently, the effect and mechanism of Cathepsin B (CTSB) in inducing apoptosis in HK-2 cells were observed and verified.ResultsThe renal injury scores, serum creatinine, blood urea nitrogen, and kidney injury molecule 1 were higher in septic mice than in non-septic mice. The proteomic analysis identified a total of 449 differentially expressed proteins (DEPs). GO and KEGG analysis showed that DEPs were mostly enriched in lysosomal-related cell structures and pathways. CTSB and MAPK were identified as key proteins in S-AKI. Electron microscopy observed enlarged lysosomes, swelled and ruptured mitochondria, and cytoplasmic vacuolization in CLP group. TUNEL staining and CTSB activity test showed that the apoptosis and CTSB activity were higher in CLP group than in control group. In HK-2 cell injury model, the CTSB activity and mRNA expression were increased in LPS-treated cells. Acridine orange staining showed that LPS caused lysosomal membrane permeabilization (LMP). CA074 as an inhibitor of CTSB could effectively inhibit CTSB activity. CCK8 and Annexin V/PI staining results indicated that CA074 reversed LPS-induced apoptosis of HK-2 cells. The JC-1 and western blot results showed that LPS inhibited mitochondrial membrane potential and activated mitochondrial apoptosis pathway, which could be reversed by CA074.ConclusionsLMP and CTSB contribute to pathogenesis of S-AKI. LPS treatment induced HK-2 cell injury by activating mitochondrial apoptosis pathway. Inhibition of CTSB might be a new therapeutic strategy to alleviate sepsis-induced acute kidney injury.https://www.frontiersin.org/articles/10.3389/fimmu.2022.1053754/fulllysosomal membrane permeabilizationCA074mitochondrial apoptosis pathwayS-AKIcathepsin B (CTSB) |
| spellingShingle | Yuting Wang Wenjie Xi Xinyi Zhang Xinwen Bi Boyang Liu Xiaoming Zheng Xinjin Chi CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway Frontiers in Immunology lysosomal membrane permeabilization CA074 mitochondrial apoptosis pathway S-AKI cathepsin B (CTSB) |
| title | CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway |
| title_full | CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway |
| title_fullStr | CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway |
| title_full_unstemmed | CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway |
| title_short | CTSB promotes sepsis-induced acute kidney injury through activating mitochondrial apoptosis pathway |
| title_sort | ctsb promotes sepsis induced acute kidney injury through activating mitochondrial apoptosis pathway |
| topic | lysosomal membrane permeabilization CA074 mitochondrial apoptosis pathway S-AKI cathepsin B (CTSB) |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1053754/full |
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