›› 2012, Vol. 32 ›› Issue (2): 133-.doi: 10.3969/j.issn.1674-8115.2012.02.002

• Original article (Basic research) • Previous Articles     Next Articles

Mechanism of inhibitory effect of insulin-like growth factor binding protein-related protein 1 on retinal angiogenesis

SUN Tao, ZHU Bi-jun, XU Xun, SU Li, GU Qing, XU Lin   

  1. Department of Ophthalmology, the First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China
  • Online:2012-02-28 Published:2012-02-28
  • Supported by:

    State Key Program of National Natural Science Foundation of China, 30930097；National Natural Science Foundation for Young Scholars of China, 30801269；Doctoral Innovation Fund of Shanghai Jiaotong University School of Medicine, BXJ0935；Shanghai Key Laboratory for Ocular Fundus Diseases Foundation, 07Z22911

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Abstract:

Objective To investigate the mechanism of inhibitory effect of insulin-like growth factor binding protein-related protein (IGFBP-rP1) on vascular endothelial growth factor (VEGF)-induced retinal angiogenesis in vitro. Methods Retina-choroid endothelial cell line (RF/6A) of macaque was cultured and starved for 24 h. Then RF/6A cells were divided into control group and 50, 100 and 200 ng/mL IGFBP-rP1 groups, and flow cytometry was used to detect the apoptotic ratios of RF/6A cells. The starved RF/6A cells were also divided into control group, 10 ng/mL VEGF group and 50, 100 and 200 ng/mL IGFBP-rP1+10 ng/mL VEGF groups, the expression of B-Raf protein was detected with immunocytochemical staining, and the activation of Caspase 3 was determined by spectrophotometric method. Results Flow cytometry revealed that there was no significant difference in apoptotic ratios of RF/6A cells between control group and 50, 100 and 200 ng/mL IGFBP-rP1 groups (P>0.05). Immunocytochemical detection and Caspase 3 activation detection demonstrated that the average optical density of RF/6A cells in 10 ng/mL VEGF group was significantly higher than that in control group (P<0.05), D405 in 10 ng/mL VEGF group was significantly lower than that in control group (P<0.05), the average optical density of RF/6A cells in 50, 100 and 200 ng/mL IGFBP-rP1+10 ng/mL VEGF groups was significantly lower than that in 10 ng/mL VEGF group (P<0.05), D405 in 50, 100 and 200 ng/mL IGFBP-rP1+10 ng/mL VEGF groups was significantly higher than that in 10 ng/mL VEGF group (P<0.05), and there were significant differences in average optical density of RF/6A cells and D405 among 50, 100 and 200 ng/mL IGFBP-rP1+10 ng/mL VEGF groups (P<0.05). Conclusion IGFBPrP1 inhibits the biologic proceedings induced by VEGF during retinal angiogenesis by intervention of expression of BRaf protein and upregulation of Caspase 3 activity.

Key words: insulin-like growth factor binding protein-related protein 1, vascular endothelial growth factor, RF/6A cell, B-Raf, Caspase 3