姜黄素通过AKT/HIF-1α/VEGF信号通路在体外抑制脉络膜新生血管的机制
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国家自然科学基金资助项目(No.81574032)


Mechanism study of Curcumin on inhibiting experimental choroidal neovascularization in vitro by AKT/HIF-1α/VEGF pathway
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National Natural Science Foundation of China(No.81574032)

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    摘要:

    目的:探讨姜黄素在体外抑制脉络膜新生血管(CNV)生成的作用及机制。

    方法:氯化钴(CoCl2)诱导人视网膜色素上皮(ARPE-19)细胞建立化学性缺氧模型,采用CCK-8法检测姜黄素对CoCl2诱导的ARPE-19细胞活性的影响,采用RT-qPCR和Western blot检测姜黄素对CoCl2诱导的ARPE-19缺氧模型细胞内AKT、HIF-1α和VEGF mRNA及蛋白表达的影响。采用细胞划痕实验、Transwell小室迁移实验、Transwell小室侵袭实验及Matrigel基质胶管腔形成实验,观察在非接触情况下ARPE-19细胞姜黄素的条件培养液对人脐静脉内皮细胞(HUVEC)的增殖、迁移、侵袭和管腔形成的影响。

    结果:100μmol/L CoCl2可成功建立ARPE-19细胞化学缺氧模型。CoCl2在100μmol/L浓度下促进ARPE-19细胞中AKT、HIF-1α和VEGF mRNA及p-AKT、HIF-1α和VEGF蛋白的表达。姜黄素在浓度为100μmol/L时可减少ARPE-19细胞中AKT、HIF-1α和VEGF mRNA及p-AKT、HIF-1α和VEGF蛋白的表达。ARPE-19细胞姜黄素低(6.25μmol/L)、中(25μmol/L)、高剂量组(100μmol/L)条件培养液可显著抑制HUVEC细胞水平迁移; 其中高剂量组条件培养液可显著抑制HUVEC细胞垂直迁移和细胞侵袭。ARPE-19细胞姜黄素中、高剂量组条件培养液可抑制HUVEC细胞管腔形成。

    结论:姜黄素在100μmol/L对CoCl2诱导ARPE-19细胞缺氧有保护作用。姜黄素可在细胞水平抑制血管的生成。

    Abstract:

    AIM: To investigate the effect and mechanism of curcumin on inhibiting choroidal neovascularization(CNV)in vitro.

    METHODS: Human retinal pigment epithelial(ARPE-19)cells chemical hypoxia model was established by cobalt chloride(CoCl2). CCK-8 method was used to detect the effect of curcumin on the activity of ARPE-19 cells induced by CoCl2. RT-qPCR and Western blot were used to detect the expression of AKT, HIF-1α, VEGF mRNA and protein in ARPE-19 cells hypoxia model induced by CoCl2. Cell scratch test, transwell chamber migration test, transwell chamber invasion test and matrigel matrix hose lumen formation test were used to observe the effects of conditioned medium of curcumin in ARPE-19 cells on the proliferation, migration, invasion and lumen formation of human umbilical vein endothelial cells(HUVEC)in non-contact condition.

    RESULTS:Chemical hypoxia model of ARPE-19 cells can successfully establish by CoCl2 at 100μmol/L. CoCl2 at the final concentration of 100μmol/L can promote the expression of AKT, HIF-1α and VEGF mRNA and p-AKT, HIF-1α and VEGF protein in ARPE-19 cells. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF-1α and VEGF mRNA in ARPE-19 hypoxia model. Curcumin at the final concentration of 100μmol/L can reduce the expression of AKT, HIF -1α and VEGF proteins in ARPE-19 hypoxia model. The conditioned medium of low(6.25μmol/L), medium(25μmol/L)and high dose(100μmol/L)curcumin in ARPE-19 cells can significantly inhibit the level migration of HUVEC. The conditioned medium in high dose group can significantly inhibit the vertical migration and cell invasion of HUVEC. The conditioned medium of middle and high dose curcumin in ARPE-19 cells can inhibit the lumen formation of HUVEC.

    CONCLUSION:Curcumin at 100μmol/L can protect ARPE-19 cells from hypoxia induced by CoCl2. Curcumin can inhibit the formation of blood vessels at the cellular level.

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陈水龄,亢泽峰,褚文丽,等.姜黄素通过AKT/HIF-1α/VEGF信号通路在体外抑制脉络膜新生血管的机制.国际眼科杂志, 2022,22(4):541-548.

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  • 收稿日期:2021-07-04
  • 最后修改日期:2022-03-14
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  • 在线发布日期: 2022-03-24
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