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VR-10 Thrombospondin-1 Synthetic Polypeptide's Impact on Rhesus Choroid-Retinal Endothelial Cells

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机构: [1]First Peoples Hosp Yunnan Prov, Dept Ophthalmol, 157 Jinbi St, Kunming, Yunnan, Peoples R China
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关键词: Platelet reactive protein VR-10 Endothelial cell Migration Proliferation Apoptosis

摘要:
Background/Aims: This study aimed to investigate the effects of the VR-10 TSP-1 synthetic polypeptide on cytokines and the proliferation and migration of endothelial cells, as well as exploring a new method for anti-ocular neoangiogenesis. Methods: We measured the proliferation of RF/6A cells by an MTT assay and investigated the migration of RF/6A cells by a Transwell chamber assay. We examined the mRNA transcript levels of TGF-beta 2, VEGF, PEDF, Bcl-2 and FasL in RF/6A cells by RT-PCR and evaluated the expression of Fas and caspase-3 proteins in RF/6A cells by western blot analysis. Results: 1. TSP-1 (1 mu g/ml) and synthetic peptide VR-10 (0.1 mu g/ml, 1 mu g/ml and 10 mu g/ml) inhibited the proliferation of RF/6A cells in a time and dose-dependent way. 2. TSP-1 and synthetic peptide VR-10 could inhibit the migration of RF/6A cells in a Transwell chamber (P < 0.001). It was demonstrated that 10 mu g/ml synthetic peptide VR-10 had the strongest effect. 3. The expression of TGF-beta 2 mRNA in RF/6A cells increased after treatment with 1 mu g/ml TSP-1 (P < 0.0001). However, there was no significant difference between the synthetic peptide VR-10 and the control group (P > 0.05). Expression of PEDF mRNA in RF/6A cells was increased after treatment with 1 mu g/ml TSP-1 and synthetic peptide VR-10. We demonstrated that 10 mu g/ml synthetic peptide VR-10 had the strongest effect (P < 0.001). There were significant differences between groups (P < 0.001). Expression of TGF-beta 2 mRNA in RF/6A cells increased after treatment with 1 mu g/ml TSP-1 (P = 0.000). There was no significant difference between the synthetic peptide VR-10 and the control group (P > 0.05). PEDF mRNA expression in RF/6A cells decreased after 1 mu g/ml TSP-1 and synthetic peptide VR-10 therapy, among which 10 mu g/ml synthetic peptide VR-10 demonstrated the strongest effect (P < 0.001). There were significant differences between groups (P < 0.001), except for the 1 mu g/ml synthetic peptide VR-10 and 1 mu g/ml synthetic peptide VR-10 groups (P = 0.615). 4. Compared with the control group, FasL mRNA expression was significantly increased in the 10 mu g/ml synthetic peptide VR-10 treatment group; however, Bcl-2 mRNA expression was decreased. 5. Western blotting showed that RF/6A cells in the control group mainly expressed the 32 kD procaspase-3 forms. For the 10 mu g/ml synthetic peptide, VR-10 treatment group, it showed decreased expression of procaspase-3 (32 kD) and concomitant increased expression of its shorter pro apoptotic forms (20 kD). Compared with the control group, Fas protein expression significantly increased in the 10 mu g/ml synthetic peptide VR-10 treatment group. Conclusions: Synthetic peptide VR-10 had an inhibitory action on the proliferation and migration of RF/6A cells. VR-10 inhibited angiogenesis by its combined actions, which included up-regulating the expression of an anti-angiogenesis gene, namely, pigment epithelium-derived factor (PEDF), down-regulating the expression of the pro-angiogenic vascular endothelial growth factor (VEGF), and mediated endothelial cell apoptosis. (C) 2018 The Author(s). Published by S. Karger AG, Basel.

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出版当年[2018]版:
大类 | 2 区 生物
小类 | 1 区 生理学 3 区 细胞生物学
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出版当年[2017]版:
Q1 CELL BIOLOGY Q1 PHYSIOLOGY
最新[2023]版:
Q2 PHYSIOLOGY Q3 CELL BIOLOGY

影响因子: 最新[2023版] 最新五年平均 出版当年[2017版] 出版当年五年平均 出版前一年[2016版]

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第一作者机构: [1]First Peoples Hosp Yunnan Prov, Dept Ophthalmol, 157 Jinbi St, Kunming, Yunnan, Peoples R China
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通讯机构: [1]First Peoples Hosp Yunnan Prov, Dept Ophthalmol, 157 Jinbi St, Kunming, Yunnan, Peoples R China [*1]Department of ophthalmology, First people’s hospital of Yunnan Province,No. 157 of Jinbi Street, Xishan District, Kunming (China)
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