白藜芦醇对人角膜上皮细胞炎症和氧化应激损伤的保护作用
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国家自然科学基金项目(No.81670837); 天津市科技计划项目(No.20JCYBJC01450); 天津市眼科医院科技基金项目(No.YKQN1921)


Protective effects of Resveratrol on inflammation and oxidative stress damage in human corneal epithelial cells
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National Natural Science Foundation of China(No.81670837); Tianjin Science and Technology Plan Program(No.20JCYBJC01450); Science and Technology Fund Project of Tianjin Eye Hospital(No.YKQN1921)

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    摘要:

    目的:探讨白藜芦醇(RSV)对人角膜上皮细胞(HCECs)炎症和氧化应激损伤的保护作用。

    方法:用肿瘤坏死因子-α(TNF-α)诱导HCECs发生炎症反应,设对照组、TNF-α组、RSV+TNF-α组; 用H2O2诱导HCECs发生氧化应激反应,设正常组、H2O2组、RSV+H2O2组。采用MTT法检测细胞活力; RT-qPCR及酶联免疫吸附测定(ELISA)法检测相关炎症因子IL-1、IL-6、IL-8的表达; 免疫荧光染色法及Western blot法检测核因子-κB p65(NF-κB p65)的核转位; 2',7'-二氯荧光黄双乙酸盐(DCFH-DA)荧光探针法检测细胞内活性氧(ROS)水平。

    结果:在HCECs炎症反应中,RT-qPCR及ELISA结果均显示,与对照组相比,TNF-α组的HCECs IL-1、IL-6、IL-8的表达水平均显著升高。RSV预处理细胞后,以上指标较TNF-α组显著下降; 免疫荧光染色及Western blot结果均显示,TNF-α组NF-κB p65核转位增加,而RSV预处理细胞后NF-κB p65核转位受到抑制。在HCECs氧化应激反应中,MTT和DCFH-DA荧光探针染色结果分别显示,H2O2刺激使HCECs活力显著下降,HCECs内ROS的产生增多。而RSV 预处理后,细胞活力显著上升,且RSV抑制了H2O2诱导的HCECs内ROS的产生。

    结论:RSV对HCECs的炎症和氧化应激反应具有抑制作用,RSV通过抑制NF-κB通路激活以抑制炎症反应。

    Abstract:

    AIM:To investigate the protective effects of resveratrol(RSV)on inflammation and oxidative stress damage in human corneal epithelial cells(HCECs).

    METHODS: The inflammation of HCECs was induced by Tumor necrosis factor-α(TNF-α), and the experiment was divided into: control group, TNF-α group and RSV+TNF-α group. The oxidative stress response of HCECs was induced by H2O2, and they were divided into normal group, H2O2 group and RSV+ H2O2 group. MTT assay was used to detect the viability of HCECs; RT-qPCR and enzyme-linked immunosorbent assay(ELISA)methods were used to detect the expression of IL-1, IL-6 and IL-8; Immunofluorescence staining and Western blot were used to observe the nuclear translocation of NF-κB p65. 2',7'-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe was applied to detect the level of reactive oxygen species(ROS).

    RESULTS:In the inflammatory response of HCECs, RT-qPCR and ELISA showed that the expression levels of IL-1, IL-6 and IL-8 were increased significantly in the TNF-α group compared with the control group, the above indicators were lower after pretreatment of RSV than those in TNF-α group; Immunofluorescence staining and Western blot showed that the nuclear translocation of NF-κB p65 was increased in TNF-α group, while it was inhibited after pretreatment of RSV. In the oxidative stress response of HCECs, the results of MTT and DCFH-DA fluorescent probe staining showed that H2O2 significantly decreased the viability of HCECs and increased the production of ROS in HCECs. After pretreatment of RSV, cell viability increased significantly, and RSV inhibited the generation of ROS in HCECs induced by H2O2.

    CONCLUSION: RSV has an inhibitory effect on inflammation and oxidative stress damage in human corneal epithelial cells, and it has been confirmed that RSV inhibits inflammation by inhibiting the activation of the NF-κB pathway.

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朱琳,韩瑞芳,王培红,等.白藜芦醇对人角膜上皮细胞炎症和氧化应激损伤的保护作用.国际眼科杂志, 2021,21(12):2038-2043.

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  • 收稿日期:2021-03-23
  • 最后修改日期:2021-11-08
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  • 在线发布日期: 2021-11-22
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