摘要
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.
出处
《国际眼科杂志》
CAS
2006年第3期580-583,共4页
International Eye Science