Abstract:

Objective　To explore the effects of hydroxysafflor yellow A (HSYA) on the proliferation, migration, and epithelial-mesenchymal transition (EMT) of human lens epithelial cell line SRA01/04 (HLEC-SRA01/04) induced by recombinant human epidermal growth factor (rhEGF). Methods　This study was from June 2022 to September 2023. The HLEC-SRA01/04 induced by rhEGF were treated with different concentrations of HSYA solution for different durations. The proliferation status of the cells of each group was detected by the MTT assay, and the IC50 value was calculated. The cell scratch test and Transwell chamber were used to measure the migration ability of the cells in different concentration groups. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western Blot were adopted to determine the mRNA and protein expression levels of proliferating cell nuclear antigen (PCNA) and vimentin involved in EMT in the cells of different concentration groups. The measurement data were compared by the t test. Results　The MTT assay results showed that the proliferation inhibition rates were (12.1±0.6)%, (19.0±3.8)%, (31.5±2.2)%, (53.7±0.4)%, and (70.8±0.3)%, respectively, and the IC50 value was (76.520±0.954) μmol/L after treating HLECs induced by 5 μg/L rhEGF with the HSYA solution of 20, 40, 60, 80, and 100 μmol/L for 24 h; the proliferation inhibition rates were (14.3±3.7)%, (27.4±3.1)%, (42.6±2.7)%, (59.2±2.2)%, and (81.0±1.0)% , respectively, and the IC50 value was (66.094±2.508) μmol/L after treating HLECs induced by 5 μg/L rhEGF with the HSYA solution of 20, 40, 60, 80, and 100 μmol/L for 48 h, indicating significant dose-dependent effects. The cell scratch test results showed that the 24 h cell migration rates of the blank group and the 0, 20, 40, 70, and 100 μmol/L groups were (46.9±1.8)%, (90.0±1.5)%, (88.4±2.1)%, (43.3±6.6)%, (31.5±16.2)%, and (5.82±5.2)%, and the 48 h cell migration rates were (81.1±2.3)%, 100%, (95.5±0.1)%, (72.6±3.5)%, (58.5±6.1)%, and (37.4±7.1)%, respectively. The Transwell chamber results showed that the numbers of cells in the blank group and the 0, 20, 40, 70, and 100 μmol/L groups were (171.667±20.407), (290.222±24.135), (198.667±16.826), (161.222±5.981), (134.111±6.850), and (67.444±7.351), respectively; HSYA inhibited the migration of HLECs induced by rhEGF in a concentration-dependent manner. The RT-qPCR and Western Blot results proved that HSYA significantly down-regulated the mRNA and protein expressions of PCNA and Vimentin in HLECs induced by rhEGF in a dose-dependent manner. Conclusion　HSYA remarkably inhibits the proliferation, migration, and EMT of HLEC-SRA01/04 induced by rhEGF, serving as a promising drug for preventing posterior capsular opacification.

Key words:

Posterior capsular opacification, Hydroxysafflor yellow A, Recombinant human epidermal growth factor, Lens epithelial cells, Epithelial-mesenchymal transition

摘要：

目的　探讨羟基红花黄色素A（HSYA）对重组人表皮生长因子（rhEGF）诱导的人晶状体上皮细胞系SRA01/04（HLEC-SRA01/04）增殖、迁移及上皮-间质转化（EMT）的影响。方法　研究时间为2022年6月至2023年9月。不同浓度HSYA溶液处理rhEGF诱导的HLEC-SRA01/04不同时长，四甲基偶氮唑蓝（MTT）比色法检测各组细胞的增殖状态并计算半数抑制浓度值（IC50）。细胞划痕实验与Transwell小室测定不同浓度组细胞的迁移能力。逆转录荧光定量聚合链式反应法（RT-qPCR）与蛋白印迹法（Western Blot）测定不同浓度组细胞内增殖细胞核抗原（PCNA）及EMT标记物波形蛋白（Vimentin）的mRNA表达水平与蛋白含量。计量资料采用t检验。结果　MTT实验结果示：选取20、40、60、80、100 μmol/L的HSYA溶液处理5 μg/L rhEGF诱导人晶状体上皮细胞24 h后，增殖抑制率分别为（12.1±0.6）%、（19.0±3.8）%、（31.5±2.2）%、（53.7±0.4）%、（70.8±0.3）%，IC50值为（76.520±0.954）μmol/L，48 h增殖抑制率分别为（14.3±3.7）%、（27.4±3.1）%、（42.6±2.7）%、（59.2±2.2）%、（81.0±1.0）%，IC50值为（66.094±2.508）μmol/L，呈明显剂量依赖性。细胞划痕实验结果显示：空白组及0、20、40、70、100 μmol/L组的24 h细胞迁移率分别为（46.9±1.8）%、（90.0±1.5）%、（88.4±2.1）%、（43.3±6.6）%、（31.5±16.2）%、（5.82±5.2）%，48 h细胞迁移率分别为（81.1±2.3）%、100%、（95.5±0.1）%、（72.6±3.5）%、（58.5±6.1）%、（37.4±7.1）%。Transwell小室实验结果显示：空白组及0、20、40、70、100 μmol/L组的细胞数分别为（171.667±20.407）个、（290.222±24.135）个、（198.667±16.826）个、（161.222±5.981）个、（134.111±6.850）个、（67.444±7.351）个，HSYA对rhEGF诱导人晶状体上皮细胞迁移的抑制呈明显浓度依赖性。RT-qPCR法及Western Blot法结果示：HSYA可明显下调rhEGF诱导的人晶状体上皮细胞内PCNA、Vimentin及mRNA表达水平，呈剂量依赖性。结论　HSYA可明显抑制rhEGF诱导的HLEC-SRA01/04增殖、迁移及EMT，可能成为预防后发性白内障的潜在药物。

关键词:

后发性白内障, 羟基红花黄色素A, 重组人表皮生长因子, 晶状体上皮细胞, 上皮-间质转化