[关键词]
[摘要]
目的:观察曲尼司特对体外培养人视网膜色素上皮(retinal pigment epithelium,RPE)细胞增殖的影响。
方法:用不同浓度的曲尼司特0(对照组),12.5,25,50,100mg/L作用于体外培养的3~5代人RPE细胞(RPE-19细胞株),在倒置显微镜下、结合HE染色观察细胞形态,并采用细胞角蛋白CK18对细胞进行鉴定。分别在作用12,24,48h后采用四甲基偶氮唑蓝(MTT)比色法检测各组细胞抑制率,蛋白质印迹法(Western-Blot)和免疫组织化学法观察不同浓度的曲尼司特作用24h后细胞表达转化生成因子-β1(TGF-β1)、血小板衍化生长因子受体A(PDGFR-A)蛋白的情况。
结果:相同时间点不同浓度曲尼司特对细胞抑制率随浓度增加而增大,差异有统计学意义(P<0.05)。TGF-β1蛋白表达定位于细胞浆,PDGFR-A蛋白定位于胞膜和胞浆,它们的表达量都随曲尼司特浓度的增加而下调,差异有统计学意义(P<0.05)。
结论:曲尼司特可以抑制人RPE细胞的增殖,并有剂量依赖性,其机制可能与下调TGF-β1和PDGFR-A蛋白表达有关。
[Key word]
[Abstract]
AIM: TO investigate the effects of tranilast on proliferation of cultured human retinal pigment epithelial(RPE)cells
in vitro.
METHODS: To treate them with different concentrations of tranilast 0(control group), 12.5, 25, 50, 100mg/L after culturing human retinal pigment epithelial cells(hRPE-19 cell line)3-5 generations. And then to observe cell morphology under inverted microscope combined with HE staining, identify the cell by keratin CK18. Detecting inhibition rate by MTT colorimetric method after the cells were treated with different concentrations of tranilast for 12h, 24h, 48h. The RPE cells were cultured with different concentrations of tranilast for 24h, then to measure the transforming growth factor β1(TGF-β1)and platelet-derived growth factor receptor A protein(PDGFR-A)expressing by Western-Blot and immunohistochemical method.
RESULTS: The cell inhibition rate increased with the increase of concentration at the same time point(P<0.05). It shows a statistically significant difference. TGF-β1 protein expression is in cell plasma and PDGFR-A protein in the cell membrane and cytoplasm, the expression of their amount was lowered with the increase of concentration of tranilast(P<0.05). It shows a statistically significant difference.
CONCLUSION: Tranilast could inhibit RPE proliferation, and it may be connected with the decreased TGF-β1 and PDGFR-A protein expression.
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