曲安奈德对色素上皮细胞衍生因子表达的影响

摘要

Objective To observe the influence of triamcinolone acetonide (TA) on the expression of pigment epithelium-derived factor (PEDF) of human retinal pigment epithelial (RPE) cells. Methods Cultured human RPE cells (4th - 6th generations) were treated with four different concentrations of TA (40, 400, 4× 103 and 4× 104 μg/L) for three different periods (12 or 24 or 48 hours), the levels of PEDF protein in the cell culture supernatant and cell lysates were determined by Western blot. After the initial experiment, RPE cells were treated with or without tumor necrosis factor-α (TNF-α, 20 ng/ml) for 24 hours, followed by TA (400 μg/L) treatment. The levels of PEDF and phospho-p38 mitogen activated protein kinase (p-p38MAPK) protein expression in cell culture supernatant and cell lysates were measured by Western blot. Results TA-treated RPE cells had higher PEDF expression, and 400 μg/L TA group had the highest effect (F= 16.98, P＜0. 05). 400μg/L TA treatment for one, six or 24 hours, with or without TNF-α pretreatment, could all promote the PEDF expression and inhibit the p-p38MAPK protein expression (F= 16.87, 10.28; P＜0. 01). TNF-α pretreatment alone could inhibit PEDF protein expression and promote p-p38MAPK protein expression (F= 16.87, 10. 28; P＜0. 01). Conclusions TA can up-regulate the expression of PEDF, and down-regulate the expression of p-p38MAPK in the cultured human RPE cells.%目的 观察曲安奈德(TA)对人视网膜色素上皮细胞衍生因子(PEDF)表达的影响.方法 体外培养人视网膜色素上皮(RPE)细胞,取第4～6代细胞用于实验.分别以浓度为40、400、4×103、4×104μg/L的TA干预12、24、48 h后,采用蛋白质免疫印迹法(Western blot)检测培养液及细胞浆中PEDF蛋白表达水平.采用20 ng/ml的肿瘤坏死因子-α(TNF-α)预干预RPE细胞24 h后,再加入400μg/L的TA进行干预(TNF-α预处理组);分别选用20 ng/ml的TNF-α和400μg/L的TA单独干预RPE细胞作为单独干预组.分别作用1、6、24 h后,测定3组细胞培养液及细胞浆中PEDF及细胞浆中磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)的蛋白表达水平.结果 各TA浓度组RPE细胞培养液及细胞浆中PEDF蛋白表达均升高,其中400μg/L组PEDF蛋白表达最高(P＜0.05).干预1、6、24 h时,TNF-α预处理组和TA单独干预组PEDF蛋白表达均增高、p-p38MAPK蛋白表达均降低(P＜0.01);TNF-α单独干预组PEDF蛋白表达降低、p-p38MAPK蛋白表达增高(P＜0.01).结论 TA能够上调体外培养的人RPE细胞PEDF蛋白表达,下调p-p38MAPK蛋白表达.