Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization
Abstract Background The relationship between the role of VEGF and autophagy in the process of retinal angiogenesis is still unclear. In this study, we explored this issue by using the mouse retinal vascular endothelial cell (RVEC) as a model. Methods RVECs were divided into the following groups: con...
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| Format: | Article |
| Language: | English |
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BMC
2018-04-01
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| Series: | BMC Ophthalmology |
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| Online Access: | http://link.springer.com/article/10.1186/s12886-018-0774-6 |
| _version_ | 1831737993780527104 |
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| author | Rong Li Jin Tian Junhui Du Lei Zhao Yang Yao Zhaoxiang Yu Weiping Chang Rui Shi Jing Li |
| author_facet | Rong Li Jin Tian Junhui Du Lei Zhao Yang Yao Zhaoxiang Yu Weiping Chang Rui Shi Jing Li |
| author_sort | Rong Li |
| collection | DOAJ |
| description | Abstract Background The relationship between the role of VEGF and autophagy in the process of retinal angiogenesis is still unclear. In this study, we explored this issue by using the mouse retinal vascular endothelial cell (RVEC) as a model. Methods RVECs were divided into the following groups: control, hypoxia (H), 3-methyladenine (3-MA) + H, VEGF + H, 3-MA + VEGF+H, anti-VEGF antibody + H, 3-MA+ anti-VEGF antibody + H. We then examined activation of autophagy by detecting formation of autophagosomes with transmission electron microscopy (TEM) and by counting the number of green fluorescent protein-positive (GFP+) puncta in RVECs. The turnover of microtubule associated protein 1 light chain 3 B (LC3B) and VEGF were examined by western blot. Cell migratory capacity was measured with wound healing assay and transwell assay. The capillary formation assay was performed to investigate the angiogenic capacity. Results Hypoxia led to an increased number of autophagosome and of the GFP+ puncta, an increased ratio of LC3B-II/I and enhanced migratory and capillary-formation capacities of RVECs. Pre-treatment with 3-MA attenuated activation of autophagy and abrogated the enhanced cell migration and capillary formation under hypoxia. Exposure to VEGF significantly increased migratory and capillary formation capacities of RVECs under hypoxia and 3-MA decreased VEGF-induced angiogenesis without its expression. Formation of autophagosome, the number of GFP+ puncta of RVECs and expression of LC3B-II/I were both elevated in cells treated with anti-VEGF antibody and these effects were partially inhibited by 3-MA pretreatment. Conclusion Our present data may identify autophagic response as a novel target for enhancing the therapeutic efficacy of angiogenesis inhibitors. |
| first_indexed | 2024-12-21T13:13:44Z |
| format | Article |
| id | doaj.art-f1dfdd91b7b24472931f66de473edbef |
| institution | Directory Open Access Journal |
| issn | 1471-2415 |
| language | English |
| last_indexed | 2024-12-21T13:13:44Z |
| publishDate | 2018-04-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Ophthalmology |
| spelling | doaj.art-f1dfdd91b7b24472931f66de473edbef2022-12-21T19:02:48ZengBMCBMC Ophthalmology1471-24152018-04-0118111210.1186/s12886-018-0774-6Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularizationRong Li0Jin Tian1Junhui Du2Lei Zhao3Yang Yao4Zhaoxiang Yu5Weiping Chang6Rui Shi7Jing Li8Department of Ophthalmology, the First Affiliated Hospital of Xi’an Medical UniversityDepartment of Ophthalmology, the Weinan Central HospitalDepartment of Ophthalmology, Xi’an Ninth Hospital Affiliated to Medical College of Xi’an Jiaotong UniversityDepartment of Molecular Physiology and Biophysics, Holden Comprehensive Cancer Center, University of Iowa Carver College of MedicineDepartment of Central laboratory, The First Affiliated Hospital of Xi’an Medical UniversityDepartment of General surgery, The First Affiliated Hospital of Xi’an Medical UniversityDepartment of General surgery, The First Affiliated Hospital of Xi’an Medical UniversityDepartment of Ophthalmology, Shaanxi Provincial People’s HospitalDepartment of Ophthalmology, Shaanxi Provincial People’s HospitalAbstract Background The relationship between the role of VEGF and autophagy in the process of retinal angiogenesis is still unclear. In this study, we explored this issue by using the mouse retinal vascular endothelial cell (RVEC) as a model. Methods RVECs were divided into the following groups: control, hypoxia (H), 3-methyladenine (3-MA) + H, VEGF + H, 3-MA + VEGF+H, anti-VEGF antibody + H, 3-MA+ anti-VEGF antibody + H. We then examined activation of autophagy by detecting formation of autophagosomes with transmission electron microscopy (TEM) and by counting the number of green fluorescent protein-positive (GFP+) puncta in RVECs. The turnover of microtubule associated protein 1 light chain 3 B (LC3B) and VEGF were examined by western blot. Cell migratory capacity was measured with wound healing assay and transwell assay. The capillary formation assay was performed to investigate the angiogenic capacity. Results Hypoxia led to an increased number of autophagosome and of the GFP+ puncta, an increased ratio of LC3B-II/I and enhanced migratory and capillary-formation capacities of RVECs. Pre-treatment with 3-MA attenuated activation of autophagy and abrogated the enhanced cell migration and capillary formation under hypoxia. Exposure to VEGF significantly increased migratory and capillary formation capacities of RVECs under hypoxia and 3-MA decreased VEGF-induced angiogenesis without its expression. Formation of autophagosome, the number of GFP+ puncta of RVECs and expression of LC3B-II/I were both elevated in cells treated with anti-VEGF antibody and these effects were partially inhibited by 3-MA pretreatment. Conclusion Our present data may identify autophagic response as a novel target for enhancing the therapeutic efficacy of angiogenesis inhibitors.http://link.springer.com/article/10.1186/s12886-018-0774-6AutophagyAngiogenesisRetinal neovascularizationHypoxiaVEGF |
| spellingShingle | Rong Li Jin Tian Junhui Du Lei Zhao Yang Yao Zhaoxiang Yu Weiping Chang Rui Shi Jing Li Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization BMC Ophthalmology Autophagy Angiogenesis Retinal neovascularization Hypoxia VEGF |
| title | Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization |
| title_full | Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization |
| title_fullStr | Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization |
| title_full_unstemmed | Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization |
| title_short | Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization |
| title_sort | manipulation of autophagy a novelly potential therapeutic strategy for retinal neovascularization |
| topic | Autophagy Angiogenesis Retinal neovascularization Hypoxia VEGF |
| url | http://link.springer.com/article/10.1186/s12886-018-0774-6 |
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