葛根素抑制糖基化终产物诱导的人视网膜色素上皮细胞增殖及低氧诱导因子-1α的表达

Puerarin inhibits the proliferation of human retinal pigment epithelial cells and the expression of hypoxia-inducible factor-1α in human RPE cells induced by advanced glycation end products

目的:研究葛根素是否抑制糖基化终产物(AGEs)诱导的人视网膜色素上皮(RPE)细胞增殖及低氧诱导因子-1α(HIF-1α)的表达。方法:应用MTT比色法检测不同浓度(0,0.01,0.1,1g/L)的葛根素对糖基化终产物(AGEs)诱导的人RPE细胞增殖的影响;应用免疫组织化学法和Western-blot分析法检测不同浓度(0,0.01,0.1,1g/L)葛根素对100mg/L的AGEs作用于人RPE细胞24h后HIF-1α的表达,并以计算机图像分析仪进行分析。结果:MTT比色法显示,不同浓度(0,0.01,0.1,1g/L)的葛根素对AGEs诱导的人RPE细胞增殖具有不同程度的抑制作用,抑制率分别为5.7%,24.1%,30.7%;不同浓度(0.01,0.1,1g/L)的葛根素与100mg/LAGEs共同作用于RPE细胞24h,免疫组织化学法检测显示,与AGEs组比较,0.01g/L以上浓度葛根素均能显著抑制AGEs诱导的RPE细胞HIF-1α的表达,0.01g/L葛根素使HIF-1α表达降低了10.2%,0.1g/L葛根素则降低了38.9%,1g/L葛根素则达到53.4%。Western-blot分析显示,与AGEs组比较,0.01g/L以上浓度葛根素均能显著抑制AGEs诱导的RPE细胞HIF-1α的表达,随着葛根素浓度的增加,HIF-1α免疫印迹带逐渐减弱,0.01g/L葛根素使HIF-1α表达降低了11.0%,0.1g/L葛根素则降低了40.3%,1g/L葛根素则达到54.4%。结论:葛根素可显著抑制AGEs诱导的人RPE细胞的增殖及HIF-1α的表达。

AIM： To investigate whether puerarin inhibits the proliferation of human retinal pigment epithelial （RPE） cells and the expression of hypoxia-inducible factor-1α （HIF-1α） in human RPE cells induced by advanced glycation end products （AGEs）. METHODS： 1.The tetrazolium dyereduction assay （MTT） was used to determine the effect of puerarin at four concentrations （0, 0.01, 0.1, 1g/L） on human RPE cell proliferation induced by AGEs. 2. Immunohistochemical staining and Western Blot analysis were performed to detect the effect of puerarin on the expression of hypoxia-inducible factor-1α at four concentrations （0, 0.01, 0.1, 1g/L） of puerarin and the same concentration of AGEs （100mg/L） acted on RPE cells for 24h, and the picture was analyzed by computer image analysis meter. RESULTS： 1.The different concentration of puerarin significantly inhibited the proliferation of RPE cells induced by AGEs, and the inhibitory rate separately was 5.7%, 24.1% and 30.7%, respectively. 2. The different concentration of puerarin and the same concentration of AGEs （100mg/L） actedon RPE cells for 24h. Immunohistochemical staining showed the inhibitory effect of puerarin on the expression of HIF-1α induced by AGEs in RPE cells was stastistically significant at concentrations above 0.01g/L compared with the con- trol, and the expression of HIF-lo decreased by 10.2%, 38.9% and 53.4% by puerarin at 0.01, 0.1 and 1g/L, respec- tively. Western Blot analysis showed puerarin inhibited the expression of HIF-1α induced by AGEs in RPE cells at concentrations above 0.01mg/L. HIF-lo banding activity gradu- ally decreased. Compared with the control, the expression of HIF-lo decreased by 11.0%, 40.3% and 54.4% by puerarin at 0.01, 0.1 and 1g/L, respectively. CONCLUSION： Puerarin can significantly inhibit the proliferation of RPE cells and the expression of HIF-1α in human RPE cells induced by AGEs. Inhibition of HIF-1α maybe a useful strategy to treat diabetic retinopathy and puerarin maybe a potential candidate in diabetic retinopathy pharmacotherapy.