The effects of recombinant human platelet derived growth factor-BB on biological behaviours of human retinal vascular endothelial cells / 中华实验眼科杂志

ABSTRACT

Objective This study was to investigate the role of recombinant human platelet derived growth factor-BB(rhPDGF-BB) in the proliferation and migration of human retinal vascular endothelial cells (hRVECs).Methods hRVECs were cultured in DMEM with 10％ fetal bovine serum.The rhPDGF-BB at 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-BB was added in the normal control group.The proliferation of the cells (absorbancy) was assayed by cell counting kit 8 (CCK8) method.Cell scratch test was employed to evaluate the relative migration area of cells (migrated acellular area/initial acellular area).The relative expression of rhPDGF-BB recepter (rhPDGF-BBR) mRNA 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 hRVECs grew well and a expressing band according with rhPDGF-BBR prime was displayed.The absorbancy values of thecells 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±0.10,0.38±0.09,0.55±0.06 and 0.61±0.05 at 24 hours after culture,and those at 48 hours were 0.75±0.06,0.81 ±0.02,0.87±0.02 and 0.98±0.02,showing significant differences among the groups (Fgroup =16.283,P =0.000;Ftime =209.129,P=0.000),and the relative migrated areas was depended upon the rhPDGF-BB dose and time.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 ng/ml,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 mRNAt =3.900,4.014,both at P ＜ 0.05;VEGF mRNAt =6.940,7.210,both at P ＜ 0.05).Conclusions rhPDGF-BB/rhPDGF-BBR signal promotes the proliferation and migration of hRVECs probably by up-regulating the expressions of integrin and VEGF.