方法：利用200μmol/L氯化钴(cobalt chloride， CoCl₂)建立体外RPE细胞的化学缺氧模型。不同浓度的Hsp90抑制剂17-AAG预处理RPE细胞1h后给予12h缺氧处理。实验分为缺氧对照组、二甲基亚砜(DMSO)对照组和17-AAG预处理组。其中17-AAG预处理组根据浓度不同又分为6组：0.01，0.10，0.50，1.00，5.00，10.00μmol/L。应用RT-PCR和Western blot 方法检测ILK表达变化情况。

结果：缺氧对照组、DMSO对照组和17-AAG预处理组ILK mRNA与内参β-actin mRNA光密度比值分别为1.32±0.04，1.29±0.03，0.93±0.06，0.70±0.05，0.53±0.03，0.44±0.04，0.32±0.04，0.30±0.03； ILK 蛋白与内参β-actin蛋白光密度比值分别为2.16±0.04，2.13±0.04，1.65±0.04，1.13±0.05，0.74±0.03，0.41±0.06，0.35±0.04，0.35±0.03。与缺氧对照组比较，17-AAG预处理组ILK mRNA和蛋白的表达明显降低(P<0.05)，呈浓度依赖性。

结论：Hsp90抑制剂17-AAG能够降低缺氧条件下RPE细胞ILK的表达。

METHODS: RPE cells were cultured with 200μmol/L cobalt chloride(CoCl₂)for 12 hours to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 hour prior to hypoxic insult. Experiment was divided into hypoxic control group, Dimethyl sulfoxide(DMSO)control group and 17-AAG pretreatment group(0.01, 0.10, 0.50, 1.00, 5.00 and 10.00μmol/L). RT-PCR and Western blot analysis were used to examine the expression of ILK in cultured RPE cells.

RESULTS: The density ratios of ILK mRNA and β-actin mRNA of hypoxic control group, DMSO control group and 17-AAG pretreatment groups were 1.32±0.04,1.29±0.03,0.93±0.06,0.70±0.05,0.53±0.03,0.44±0.04,0.32±0.04,0.30±0.03; and the density ratios of ILK protein and β-actin protein were 2.16±0.04, 2.13±0.04, 1.65±0.04, 1.13±0.05, 0.74±0.03, 0.41±0.06, 0.35±0.04, 0.35±0.03. The ILK expression in 17-AAG pretreated groups were inhibited compared to hypoxic control group(P < 0.05), and the decrease was in concentration-dependent manner.

CONCLUSION: Hsp90 inhibitor 17-AAG was effective to inhibit the expression of ILK induced by hypoxia in RPE cells.