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目的 观察血小板源性生长因子 (platelet- derived growth factor,PDGF)对培养的人视网膜色素上皮 (retinal pigment epithelium ,RPE)细胞表达α-平滑肌肌动蛋白 (α- smooth muscle actin,α- SMA)的影响。 方法 将体外培养的第 4～ 6代人 RPE细胞分为无血清组 [改良 Eagle培养液 (Delbecco′s modi-fied Eagle′s medium ,DMEM)组 ]和含有 2 0 g/ L小牛血清的 DMEM组 (2 % DMEM组 )。每组均分别加入不同浓度 (0 ,1,5 0 ng/ ml)的 PDGF,采用免疫荧光法定性、定量观察 PDGF对人 RPE细胞表达α- SMA的影响。 结果 DMEM组中 ,无 PDGF刺激时 ,α- SMA表达阳性细胞数比率约为 4 0 %～ 5 0 % ,荧光强度的平均值为 8.0 8;加入 PDGF(1ng/ ml,5 0 ng/ ml)刺激后 ,α- SMA表达阳性细胞数的比率及荧光强度的平均值分别为 80 %、12 .35和 90 %、17.2 3。 2 % DMEM组无 PDGF刺激时 ,α- SMA表达阳性细胞数比率约为85 % ,荧光强度的平均值为 14 .79;经 1ng/ ml和 5 0 ng/ ml的 PDGF刺激后 ,α- SMA表达阳性细胞数的比率及荧光强度的平均值分别为 95 %、16 .2 8和 10 0 %、2 1.36。 2 % DMEM组经 5 0 mg/ ml PDGF刺激后的荧光强度约为 DMEM组无 PDGF刺激的时 2 .7倍 ,为 2 % DMEM组无 PDGF刺激时的 1.5倍。 结
Objective To observe the effect of platelet-derived growth factor (PDGF) on the expression of α-smooth muscle actin (α-SMA) in cultured human retinal pigment epithelium (RPE) influences. Methods The 4th to 6th generation human RPE cells cultured in vitro were divided into serum-free group [DMEM group] and DMEM group containing 20 g / L bovine serum DMEM group (2% DMEM group). The PDGF of different concentrations (0, 150 ng / ml) were added to each group, and the effect of PDGF on α-SMA expression of human RPE cells was quantitatively determined by immunofluorescence. Results The percentage of α-SMA positive cells in DMEM group was 40% ~ 50% and the average fluorescence intensity was 8.08 in the absence of PDGF stimulation. PDGF (1ng / ml, 50 ng / ml) After stimulation, the ratio of α-SMA positive cells and the average fluorescence intensity were 80%, 12.35 and 90%, 17.23, respectively. The percentage of α-SMA positive cells in 2% DMEM group was about 85% without PDGF stimulation, and the average fluorescence intensity was 14.79. After stimulated with 1ng / ml and 50ng / ml PDGF, α-SMA The average number of positive cells and the average fluorescence intensity were 95%, 16.28 and 100%, respectively, 2 1.36. The fluorescence intensity of 2% DMEM group stimulated by 50 mg / ml PDGF was about 1.7 times higher than that of DMEM group without PDGF stimulation, which was 1.5 times higher than that of 2% DMEM group without PDGF stimulation. Knot