Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling
Abstract Background In the central nervous system (CNS), connexin 43 (Cx43) is mainly expressed in astrocytes and regulates astrocytic network homeostasis. Similar to Cx43 overexpression, abnormal excessive opening of Cx43 hemichannels (Cx43Hcs) on reactive astrocytes aggravates the inflammatory res...
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-10-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12974-020-01978-z |
| _version_ | 1818117058555019264 |
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| author | Hailong Yu Xiang Cao Wei Li Pinyi Liu Yuanyuan Zhao Lilong Song Jian Chen Beilei Chen Wenkui Yu Yun Xu |
| author_facet | Hailong Yu Xiang Cao Wei Li Pinyi Liu Yuanyuan Zhao Lilong Song Jian Chen Beilei Chen Wenkui Yu Yun Xu |
| author_sort | Hailong Yu |
| collection | DOAJ |
| description | Abstract Background In the central nervous system (CNS), connexin 43 (Cx43) is mainly expressed in astrocytes and regulates astrocytic network homeostasis. Similar to Cx43 overexpression, abnormal excessive opening of Cx43 hemichannels (Cx43Hcs) on reactive astrocytes aggravates the inflammatory response and cell death in CNS pathologies. However, the role of excessive Cx43Hc opening in intracerebral hemorrhage (ICH) injury is not clear. Methods Hemin stimulation in primary cells and collagenase IV injection in C57BL/6J (B6) mice were used as ICH models in vitro and in vivo. After ICH injury, the Cx43 mimetic peptide Gap19 was used for treatment. Ethidium bromide (EtBr) uptake assays were used to measure the opening of Cx43Hcs. Western blotting and immunofluorescence were used to measure protein expression. qRT-PCR and ELISA were used to determine the levels of cytokines. Coimmunoprecipitation (Co-IP) and the Duolink in situ proximity ligation assay (PLA) were applied to measure the association between proteins. Results In this study, Cx43 expression upregulation and excessive Cx43Hc opening was observed in mice after ICH injury. Delayed treatment with Gap19 significantly alleviated hematoma volume and neurological deficits after ICH injury. In addition, Gap19 decreased inflammatory cytokine levels in the tissue surrounding the hematoma and decreased reactive astrogliosis after ICH injury in vitro and in vivo. Intriguingly, Cx43 transcriptional activity and expression in astrocytes were significantly increased after hemin stimulation in culture. However, Gap19 treatment downregulated astrocytic Cx43 expression through the ubiquitin-proteasome pathway without affecting Cx43 transcription. Additionally, our data showed that Gap19 increased Yes-associated protein (YAP) nuclear translocation. This subsequently upregulated SOCS1 and SOCS3 expression and then inhibited the TLR4-NFκB and JAK2-STAT3 pathways in hemin-stimulated astrocytes. Finally, the YAP inhibitor, verteporfin (VP), reversed the anti-inflammatory effect of Gap19 in vitro and almost completely blocked its protective effects in vivo after ICH injury. Conclusions This study provides new insight into potential treatment strategies for ICH injury involving astroglial Cx43 and Cx43Hcs. Suppression of abnormal astroglial Cx43 expression and Cx43Hc opening by Gap19 has anti-inflammatory and neuroprotective effects after ICH injury. |
| first_indexed | 2024-12-11T04:32:22Z |
| format | Article |
| id | doaj.art-be0775bc031840d09dc8b6d4b2105ec8 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-12-11T04:32:22Z |
| publishDate | 2020-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-be0775bc031840d09dc8b6d4b2105ec82022-12-22T01:20:50ZengBMCJournal of Neuroinflammation1742-20942020-10-0117111910.1186/s12974-020-01978-zTargeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signalingHailong Yu0Xiang Cao1Wei Li2Pinyi Liu3Yuanyuan Zhao4Lilong Song5Jian Chen6Beilei Chen7Wenkui Yu8Yun Xu9Affiliated of Drum Tower Hospital, Medical school of Nanjing UniversityAffiliated of Drum Tower Hospital, Medical school of Nanjing UniversityDepartment of Neurology, Northern Jiangsu People’s HospitalAffiliated of Drum Tower Hospital, Medical school of Nanjing UniversityDepartment of Neurology, Northern Jiangsu People’s HospitalDepartment of Neurology, Northern Jiangsu People’s HospitalAffiliated of Drum Tower Hospital, Medical school of Nanjing UniversityClinical Medical College of Yangzhou UniversityAffiliated of Drum Tower Hospital, Medical school of Nanjing UniversityAffiliated of Drum Tower Hospital, Medical school of Nanjing UniversityAbstract Background In the central nervous system (CNS), connexin 43 (Cx43) is mainly expressed in astrocytes and regulates astrocytic network homeostasis. Similar to Cx43 overexpression, abnormal excessive opening of Cx43 hemichannels (Cx43Hcs) on reactive astrocytes aggravates the inflammatory response and cell death in CNS pathologies. However, the role of excessive Cx43Hc opening in intracerebral hemorrhage (ICH) injury is not clear. Methods Hemin stimulation in primary cells and collagenase IV injection in C57BL/6J (B6) mice were used as ICH models in vitro and in vivo. After ICH injury, the Cx43 mimetic peptide Gap19 was used for treatment. Ethidium bromide (EtBr) uptake assays were used to measure the opening of Cx43Hcs. Western blotting and immunofluorescence were used to measure protein expression. qRT-PCR and ELISA were used to determine the levels of cytokines. Coimmunoprecipitation (Co-IP) and the Duolink in situ proximity ligation assay (PLA) were applied to measure the association between proteins. Results In this study, Cx43 expression upregulation and excessive Cx43Hc opening was observed in mice after ICH injury. Delayed treatment with Gap19 significantly alleviated hematoma volume and neurological deficits after ICH injury. In addition, Gap19 decreased inflammatory cytokine levels in the tissue surrounding the hematoma and decreased reactive astrogliosis after ICH injury in vitro and in vivo. Intriguingly, Cx43 transcriptional activity and expression in astrocytes were significantly increased after hemin stimulation in culture. However, Gap19 treatment downregulated astrocytic Cx43 expression through the ubiquitin-proteasome pathway without affecting Cx43 transcription. Additionally, our data showed that Gap19 increased Yes-associated protein (YAP) nuclear translocation. This subsequently upregulated SOCS1 and SOCS3 expression and then inhibited the TLR4-NFκB and JAK2-STAT3 pathways in hemin-stimulated astrocytes. Finally, the YAP inhibitor, verteporfin (VP), reversed the anti-inflammatory effect of Gap19 in vitro and almost completely blocked its protective effects in vivo after ICH injury. Conclusions This study provides new insight into potential treatment strategies for ICH injury involving astroglial Cx43 and Cx43Hcs. Suppression of abnormal astroglial Cx43 expression and Cx43Hc opening by Gap19 has anti-inflammatory and neuroprotective effects after ICH injury.http://link.springer.com/article/10.1186/s12974-020-01978-zIntracerebral hemorrhage injuryConnexin 43HemichannelsReactive astrocyteYAP |
| spellingShingle | Hailong Yu Xiang Cao Wei Li Pinyi Liu Yuanyuan Zhao Lilong Song Jian Chen Beilei Chen Wenkui Yu Yun Xu Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling Journal of Neuroinflammation Intracerebral hemorrhage injury Connexin 43 Hemichannels Reactive astrocyte YAP |
| title | Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling |
| title_full | Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling |
| title_fullStr | Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling |
| title_full_unstemmed | Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling |
| title_short | Targeting connexin 43 provides anti-inflammatory effects after intracerebral hemorrhage injury by regulating YAP signaling |
| title_sort | targeting connexin 43 provides anti inflammatory effects after intracerebral hemorrhage injury by regulating yap signaling |
| topic | Intracerebral hemorrhage injury Connexin 43 Hemichannels Reactive astrocyte YAP |
| url | http://link.springer.com/article/10.1186/s12974-020-01978-z |
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