南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (5): 722-728.doi: 10.12122/j.issn.1673-4254.2021.05.13

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姜黄素通过 Wnt/β-catenin 信号通路诱导人晶状体上皮细胞的凋亡和细胞周期阻滞

朱梦云，崔双慧，郝泽宇，王文锐，杨清玲，陈昌杰，王剑锋，周 琦   

  • 出版日期:2021-05-20 发布日期:2021-06-11

Curcumin induces human lens epithelial cell apoptosis and cell cycle arrest by inhibiting Wnt/β-catenin signaling pathway

  • Online:2021-05-20 Published:2021-06-11

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摘要： 目的 探讨姜黄素对人晶状体上皮细胞株SRA01/04（HLEC-SRA01/04）细胞周期和凋亡的影响及其相关分子机制，为姜黄素治疗和预防后发性白内障提供理论依据。方法 体外培养HLEC-SRA01/04细胞，加入终浓度分别为0 μmol/L（对照组）以 及20、40和60 μmol/L的姜黄素处理孵育24 h或48 h后，MTT法检测各组细胞的增殖抑制率；流式细胞仪检测HLEC-SRA01/04细胞周期、线粒体膜电位和细胞凋亡率；Western blot法检测姜黄素对HLEC-SRA01/04细胞中caspase-9、caspase-3、Bcl-2、Bax、Cyclin B1、CDK1、β-catenin、c-myc、Cyclin D1蛋白水平表达的影响。结果 MTT显示：随着姜黄素的浓度逐渐增加，HLEC-SRA01/04细胞较对照组抑制率逐渐增高（P<0.001）；随着作用时间的延长，各实验组HLEC-SRA01/04细胞增殖抑制率均逐渐增高（P<0.001）；流式细胞仪结果显示：各组中HLEC-SRA01/04细胞凋亡率和细胞G2/M期比例随姜黄素浓度增加逐渐增高，而线粒体膜电位随着姜黄素浓度增加逐渐下降（P<0.001）。Western blot 实验显示：随着姜黄素浓度的增加，HLEC- SRA01/04细胞中caspase-3、caspase-9和Bax的表达量逐渐增高，Bcl-2、Cyclin B1、CDK1和β-catenin的表达量逐渐降低，同时Wnt/β-catenin信号通路下游靶蛋白Cyclin D1和c-myc表达量也逐渐降低。结论 姜黄素可能通过抑制Wnt/β-catenin信号通路进而抑制HLEC-SRA01/04细胞增殖，并将细胞周期阻滞在G2/M期，从而诱导其凋亡。

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Abstract: Objective To investigate the effect of curcumin on cell cycle and apoptosis of human lens epithelial cells and the possible molecular mechanism. Methods Cultured human lens epithelial cell line HLEC-SRA01/04 was treated with 20, 40 and 60 μmol/L curcumin for 24 or 48 h. The cell proliferation inhibition rate was determined using MTT assay, and the changes in cell cycle, mitochondrial membrane potential and apoptosis rate were analyzed with flow cytometry. Western blotting was used to detect the expression levels of caspase-9, caspase-3, Bcl-2, Bax, cyclin B1, CDK1, β-catenin, c-myc, and cyclin D1 in the cells. Results Curcumin concentration- and time-dependently inhibited the proliferation of in HLEC-SRA01/04 cells as compared with the control cells (P<0.05). Flow cytometric analysis showed that curcumin significantly increased apoptosis rate and cell percentage in G2/M phase and lowered mitochondrial membrane potential of HLEC-SRA01/04 cells in a concentration dependent manner (P<0.05). The results of Western blotting showed that curcumin also concentration-dependently increased the cellular expressions of caspase-3, caspase-9 and Bax and lowered the expressions of Bcl-2, cyclin B1, CDK1 and β-catenin along with the downstream proteins cyclin D1 and c-myc in the Wnt/β-catenin signaling pathway (P<0.05). Conclusion Curcumin inhibits the proliferation of HLEC-SRA01/04 cells possibly by inhibiting the Wnt/β-catenin signaling pathway and causing cell cycle arrest to induce cell apoptosis.

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