Mechanism study of Curcumin on inhibiting experimental choroidal neovascularization in vitro by AKT/HIF-1α/VEGF pathway

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.

Key words:choroidal neovascularization; Curcumin; vascular endothelial growth factor(VEGF); hypoxia-inducible factor-1α(HIF-1α); protein kinase B(AKT); human retinal pigment epithelial cells; human umbilical vein endothelial cells