姜黄素对人晶状体上皮细胞增生的抑制作用

Inhibitory effect of curcumin on proliferation of human lens epithelial cell

目的探讨姜黄素(curcumin,CUR)对人晶状体上皮细胞株SRA01/04(human lens epithelial cell line SRA01/04,HLECSRA01/04)增生的抑制作用及其剂量-效应与时间-效应的关系。方法MTT比色法分别测定不同浓度的CUR作用于HLEC不同时间后的细胞生长抑制率;不同浓度的CUR作用于HLEC24h后,流式细胞术检测HLEC的细胞凋亡率;DAPI免疫荧光染色观察HLEC的形态学改变;免疫组织化学检测HLEC的caspase-3蛋白表达情况。结果MTT比色法实验结果显示,随药物浓度增加和作用时间延长,HLEC生长抑制率增加(P<0.05)。CUR作用于HLEC24h后,流式细胞术检测表明,细胞凋亡率随药物浓度增加而明显增加(P<0.05);DAPI免疫荧光染色可见细胞核固缩、溶解碎裂、产生凋亡小体,而对照组细胞形态规则,细胞核呈现均匀的荧光染色;免疫组织化学可见,用药组caspase-3蛋白的表达随CUR浓度增加而增强(P<0.05)。结论CUR对HLEC有抑制增生和诱导凋亡作用,且呈明显的剂量-效应与时间-效应关系,可望成为后发性白内障的防治药物。

Objective To investigate the inhibitory effects of curcumin（CUR） on proliferation of human lens epithelial cell line SRA01/04（HLEC SRA01/04）,and its dose-dependent,time-dependent manner.Methods HLEC were incubated with CUR at different concentrations for different hours,and then calculated the inhibitive ratio of growth through MTT assay.After HLEC were incubated with CUR at different concentrations for 24 hours,the apoptosis of cell was assayed by flow cytometric analysis（FCM）, Morphological changes of HLEC were observed with 4,6-diamidino-2-phenylindole dihydro-chloride（DAPI） immunofluorescence staining, The expression level of caspase-3 was performed by immunohistochemical analysis.Results MTT assay showed that the inhibitory effect of CUR on HLEC was markedly correlated with the dose and time（P〈0.05）.After treatment with CUR at different concentrations for 24 hours,by FCM assay the apoptosis of cell was obviously observed in the treated groups,the ratio ascended depending on the dose（P〈0.05）.Characteristic apoptotic features such as chromatin condensation,nuclear fragmentation,cellular shrinkage and apoptotic bodies were examined under fluorescence microscope.Compared with control group,caspase-3 expression increased depending on the dose by immunohistochemistry（P〈0.05）.Conclusion CUR can inhibit proliferation and induce apoptosis in HLEC at a dose-dependent and time-dependent manner.CUR may become a potential drug to prevent and cure after cataract.