| Background:Retinal edema,hemorrhage and detachment caused by retinal neovascularization is an important cause of retinal damage.A variety of cytokines are associated with the formation of new blood vessels,and the mechanism is not fully clear.To explore the possible pathogenesis and to find effective intervention method are the hotspot in the current research field of ophthalmology.rhPDGF-BB(recombinant human platelet derived growth factor-BB)played an important role in development of retinal neovascularization,but its effects on HRECs(Human retinal microvascular endothelial cells)function was known rarely.Objective:To address the effect of rhPDGF-BB protein on the process ofretinal neovascularization,human retinal vascular endothelial cells(HRECs)was cultured in vitro with different concentration ofrhPDGF-BB,the influence of rhPDGF-BB on the proliferation,migration and of HRECs was recorded.It may provide some beneficial theoretical basis for the pathogenesis of retinal neovascularization and explore new therapeutic targets for clinical.Methods: HRECs were cultured in DMEM with 10% fetal bovine serum.TherhPDGF-BBat the concentrations of 10,50 and 200 ng/ml were added into the medium of exponential phase-growth cells for 24 and 48 hours,respectively,and no rhPDGF-BBwas added in the normal control group.The proliferation of the cells(absorbancy)was assayed by CCK8 method.Cellscratch test was employed to evaluate the relative migration area of cells(migrated acellular area/initial acellular area).The relative expression of rhPDGF-BB recepter mRNA(rhPDGF-BBR)in the cells was detected by reverse transcription PCR.The relative expression of VEGF mRNA and Integrin mRNA in the cells was detected using real-time fluorescence quantitative PCR.Results: HRECs grew well and an expressing band according with rhPDGF-BBR prime was displayed.The absorbancy values of the cells were 1.01±0.05,1.09±0.04,1.10±0.02 and 1.13±0.05 in the normal control group and 10,50,200 ng/ml rhPDGF-BB groups at 24 hours after culture,and those in the 10,50 and 200 ng/ml rhPDGF-BB groups were significantly increased in comparison with the normal control group(t=2.504,3.430,3.483,all at P<0.05).The relative migrated areas of the cells in the normal control group and 10,50,200 ng/ml rhPDGF-BB groups were(0.42±0.10)mm2,(0.38±0.09)mm2,(0.55±0.06)mm2 and(0.61±0.05)mm2 at 24 hours after culture,and those at 48 hours were(0.75±0.06)mm2,(0.81±0.02)mm2,(0.87±0.02)mm2 and(0.98±0.02)mm2,showing significant differences(Fconcentration=16.283,P=0.000;Ftime=209.129,P=0.000),and the relative migrated areas depended upon the rhPDGF-BB doses and time(all at P<0.05).The relative expressions of integrin mRNA were 1.06±0.02,1.30±0.10,1.20±0.16 and 1.27±0.08,and those of VEGF mRNA were 0.97±0.05,1.06±0.16,1.58±0.18 and 1.66±0.21 in the normal control group and 10,50,200 ng/ml rhPDGF-BB groups,respectively,and increased expressions of integrin mRNA and VEGF mRNA were found in the 50 and 200 ng/ml rhPDGF-BB groups compared with the normal control group(Integrin mRNA:t=3.900,014,both at P<0.05;VEGF mRNA:t=6.940,7.210,both at P<0.05).Conclusions: rhPDGF-BB/PDGF-BBRsignal promotes the proliferation and migration of HRECs probably by up-regulating expressions of Integrin and VEGF. |
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