Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy
Abstract Background Idiopathic pulmonary fibrosis is a chronic progressive, lethal disease in which ectopic lung fibroblast (LF) activation plays a vital part. We have previously shown that alamandine (ALA) exerts anti-fibrosis effects via the MAS-related G-protein coupled receptor D (MrgD). Here, w...
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| Format: | Article |
| Language: | English |
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BMC
2023-01-01
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| Series: | Journal of Translational Medicine |
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| Online Access: | https://doi.org/10.1186/s12967-022-03837-2 |
| _version_ | 1797952339229802496 |
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| author | Wei Wang Yue Zhang Wenhui Huang Yafei Yuan Qiaohui Hong Zhanzhan Xie Lijuan Li Yixin Chen Xu Li Ying Meng |
| author_facet | Wei Wang Yue Zhang Wenhui Huang Yafei Yuan Qiaohui Hong Zhanzhan Xie Lijuan Li Yixin Chen Xu Li Ying Meng |
| author_sort | Wei Wang |
| collection | DOAJ |
| description | Abstract Background Idiopathic pulmonary fibrosis is a chronic progressive, lethal disease in which ectopic lung fibroblast (LF) activation plays a vital part. We have previously shown that alamandine (ALA) exerts anti-fibrosis effects via the MAS-related G-protein coupled receptor D (MrgD). Here, we further investigate how it moderates transforming growth factor β1 (TGF-β1)-induced LF activation by regulating glucose metabolism and mitochondria autophagy (mitophagy). Methods In vitro, we examined glycolysis-related protein hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), and lactic acid in cells treated with TGF-β1. The oxygen consumption rate and the extracellular acidification rate were detected using Seahorse assays. Then, mitophagy was evaluated using transmission electron microscopy, mt-Keima, and the co-localization of Parkin and COX IV with LC3 and LAMP1, respectively. The autophagic degradation of HK2 and PFKFB3 was detected by 3MA and bafilomycin A1 and assessed by their co-localization with LC3 and LAMP1, respectively. The effects of ALA on LF activation markers collagen I and α-SMA were detected. The effects of ALA on glucose metabolism, mitophagy, and the activation of LF were also investigated in vivo. Results We found that the ALA/MrgD axis improved TGF-β1-mediated LF activation by repressing glycolysis by downregulating HK2 and PFKFB3 expression. Lactic acid sustained positive feedback between glycolysis and LF activation by maintaining the expression of HK2 and PFKFB3. We also showed that glycolysis enhancement resulted from blocking the autophagic degradation of HK2 and PFKFB3 while upregulated mRNA levels by TGF-β1, while all of those improved by ALA adding. Importantly, we determined that moderation of Parkin/LC3-mediated mitophagy by TGF-β1 also promotes glycolysis but is reversed by ALA. Furthermore, we proved that ALA counteracts the effects of bleomycin on HK2, PFKFB3, LC3, Parkin, and LF activation in vivo. Conclusion In this study, we show that the ALA/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy. |
| first_indexed | 2024-04-10T22:45:53Z |
| format | Article |
| id | doaj.art-5225f33069514c528508cc4ad5fadb84 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-04-10T22:45:53Z |
| publishDate | 2023-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-5225f33069514c528508cc4ad5fadb842023-01-15T12:20:16ZengBMCJournal of Translational Medicine1479-58762023-01-0121111510.1186/s12967-022-03837-2Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagyWei Wang0Yue Zhang1Wenhui Huang2Yafei Yuan3Qiaohui Hong4Zhanzhan Xie5Lijuan Li6Yixin Chen7Xu Li8Ying Meng9Department of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Emergency Medicine, Nanfang Hospital, Southern Medical UniversityDepartment of Respiratory and Critical Care Medicine, Nanfang Hospital, Southern Medical UniversityAbstract Background Idiopathic pulmonary fibrosis is a chronic progressive, lethal disease in which ectopic lung fibroblast (LF) activation plays a vital part. We have previously shown that alamandine (ALA) exerts anti-fibrosis effects via the MAS-related G-protein coupled receptor D (MrgD). Here, we further investigate how it moderates transforming growth factor β1 (TGF-β1)-induced LF activation by regulating glucose metabolism and mitochondria autophagy (mitophagy). Methods In vitro, we examined glycolysis-related protein hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), and lactic acid in cells treated with TGF-β1. The oxygen consumption rate and the extracellular acidification rate were detected using Seahorse assays. Then, mitophagy was evaluated using transmission electron microscopy, mt-Keima, and the co-localization of Parkin and COX IV with LC3 and LAMP1, respectively. The autophagic degradation of HK2 and PFKFB3 was detected by 3MA and bafilomycin A1 and assessed by their co-localization with LC3 and LAMP1, respectively. The effects of ALA on LF activation markers collagen I and α-SMA were detected. The effects of ALA on glucose metabolism, mitophagy, and the activation of LF were also investigated in vivo. Results We found that the ALA/MrgD axis improved TGF-β1-mediated LF activation by repressing glycolysis by downregulating HK2 and PFKFB3 expression. Lactic acid sustained positive feedback between glycolysis and LF activation by maintaining the expression of HK2 and PFKFB3. We also showed that glycolysis enhancement resulted from blocking the autophagic degradation of HK2 and PFKFB3 while upregulated mRNA levels by TGF-β1, while all of those improved by ALA adding. Importantly, we determined that moderation of Parkin/LC3-mediated mitophagy by TGF-β1 also promotes glycolysis but is reversed by ALA. Furthermore, we proved that ALA counteracts the effects of bleomycin on HK2, PFKFB3, LC3, Parkin, and LF activation in vivo. Conclusion In this study, we show that the ALA/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy.https://doi.org/10.1186/s12967-022-03837-2Pulmonary fibrosisFibroblast activationAlamandine/MrgD axisMitophagyAerobic glycolysis |
| spellingShingle | Wei Wang Yue Zhang Wenhui Huang Yafei Yuan Qiaohui Hong Zhanzhan Xie Lijuan Li Yixin Chen Xu Li Ying Meng Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy Journal of Translational Medicine Pulmonary fibrosis Fibroblast activation Alamandine/MrgD axis Mitophagy Aerobic glycolysis |
| title | Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| title_full | Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| title_fullStr | Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| title_full_unstemmed | Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| title_short | Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| title_sort | alamandine mrgd axis prevents tgf β1 mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy |
| topic | Pulmonary fibrosis Fibroblast activation Alamandine/MrgD axis Mitophagy Aerobic glycolysis |
| url | https://doi.org/10.1186/s12967-022-03837-2 |
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