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. 2023 May 12;13(11):1326–1345. doi: 10.1016/j.jpha.2023.05.002

Fig. 3.

Fig. 3

Dose-response effects of Fe-dihydromyricetin (DMY) nano-coordinated polymer particles (NCPs) on high glucose-induced human umbilical vein endothelial cells (HUVEC) and in vitro reactive oxygen species (ROS) scavenging ability of Fe-DMY NCPs. (A) Cell Counting Kit-8 (CCK-8) assay to detect the cell viability of high glucose-induced HUVEC after treatment with different doses of Fe-DMY NCPs for 0.5 h. (B) Dose-response curves and calculation of median effective dose (ED50) values of high glucose-induced HUVEC after 0.5 h of treatment with different doses of Fe-DMY NCPs. (C) Time-dose-activity relationship curves for different doses of Fe-DMY NCPs after 0.5, 1, 6, 12 and 24 h of treatment in high glucose-induced HUVEC. (D) Morphological changes of HUVEC after 0.5 h in normal control (NC), diabetic retinopathy (DR) control (DC), and DC + Fe-DMY NCPs (200 μg/mL) observed by inverted microscopy. (E) Fluorescence microscopy images of HUVEC stained with 2′,7′dichlorofluorescin diacetate (DCFH-DA) and 4′,6-diamidino-2-phenylindole (DAPI) after different treatments. Fe-DMY NCPs concentrations in the different groups were: DC + Fe-DMY1, 200 μg/mL; DC + Fe-DMY2, 100 μg/mL; DC + Fe-DMY3, 50 μg/mL; and DC + Fe-DMY4, 10 μg/mL. (F) Mean fluorescence integral optical density (IOD) calculated from the DCFH-DA fluorescence images quantified in Fig. 3E. (G) Fluorescence enzyme spectra measurements of ROS levels in HUVEC stained with DCFH-DA after different treatments. Fe-DMY NCPs concentrations in the different groups were: DC + Fe-DMY1, 200 μg/mL; DC + Fe-DMY2, 100 μg/mL; DC + Fe-DMY3, 50 μg/mL; and DC + Fe-DMY4, 10 μg/mL. (H) Flow cytometric quantification of HUVEC stained with DCFH-DA after different treatments. Fe-DMY NCPs concentrations in the different groups were: DC + Fe-DMY1, 200 μg/mL; DC + Fe-DMY2, 100 μg/mL; DC + Fe-DMY3, 50 μg/mL; and DC + Fe-DMY4, 10 μg/mL. (I) Mean fluorescence intensity (MFI) calculated according to Fig. 3H. Data are presented as mean ± standard error of mean, ∗∗∗∗P < 0.0001 represented the significant difference between the administered (intervention) group and the DC group, and ####P < 0.0001 represented the significant difference between the NC group and the DC group using one-way analysis of variance (ANOVA). OD: optical density.