结缔组织生长因子对视网膜色素上皮细胞增生和黏附的调节作用

目的观察重组人结缔组织生长因子(rhCTGF)对视网膜色素上皮(RPE)细胞增生和黏附的影响。方法用CTGF蛋白、多聚赖氨酸(PLL)和胶原分别包被培养板,观察CTGF促进RPE细胞黏附的作用;用四唑盐(MTT)比色试验和流式细胞仪(FCM)测定RPE细胞增生反应。结果10-30 mg/L的CTGF蛋白提高RPE细胞的贴壁黏附能力,与未包被的对照组相比,差异有统计学意义(P<0.05);30 mg/L CTGF促进RPE细胞黏附的能力与0.1 g/L胶原的作用一致,但略低于0.1 g/L PLL的作用。MTT实验显示CTGF刺激RPE细胞生长,CTGF作用24 h刺激RPE细胞增生的能力最强。FCM测定细胞周期结果显示S期百分比随着CTGF浓度增加而逐渐增加,无血清培养的对照组S期百分比为(7.5±0.4)%,60μg/LCTGF刺激24 h后,S期百分比上升至(16.1±2.0)%。结论CTGF可以促进RPE细胞的黏附和增生,在增生性玻璃体视网膜病变等眼内增生性疾病中可能有重要意义。

Modulation of adhesion and proliferation of retinal pigment epithelium cells by recombinant human connective tissue growth factor

Objective To investigate the effect of recombinant human connective tissue growth factor(rhCTGF) on adhesion and proliferation of retinal pigment epithelium(RPE) cells.Methods Microtiter plates of 96-wells were coated with CTGF,collagen protein and Poly-L-lysine(PLL),respectively,to observe the effect of CTGF on RPE cell adhesion.Proliferation in RPE cells was evaluated using MTT assay and flow cytometry.Results 10-30 mg/L CTGF protein promoted the adhesion of RPE cells,which was significantly different when compared with the control(P<0.05).CTGF at 30 mg/L enhanced adhesion of RPE cells,and the effect was the same as that of collagen in 0.1 g/L,but less than that of PLL in 0.1 g/L.MTT assay showed that CTGF promoted the proliferation of RPE cells,and the maximal effect was observed at 24 hours by 60 μg/L.FCM revealed that CTGF increased in the S phase cell percentage,and the S phase value in the control was(7.5±0.4)% and(16.1±2.0)% in 60 μg/L CTGF.Conclusion rhCTGF stimulates adhesion and proliferation of RPE cells,and CTGF may play an important role in the pathogenesis of intraocular proliferative diseases such as proliferative vitreoretinopathy.