Figures & data
Figure 1. TMZ inhibited the proliferation of HRECs induced by HG.
(a) Chemical formula for TMZ. (b) Cell Counting Kit-8 assay detected the effects of TMZ with different concentrations (0.1 µM, 1 µM and 10 µM) on HRECs viability. (c) Effects of TMZ on HRECs viability induced by HG. (d) EDU examined the effects of TMZ on HRECs proliferation induced by HG. **P < 0.01 vs. HG. TMZ, Trimetazidine; HRECs, Human retinal endothelial cells; EDU, 5-Ethynyl-2’-deoxyuridine; HG, high glucose.
Figure 2. TMZ inhibited migration, invasion, and angiogenesis of HG-induced HRECs.
(a)Wound healing and (b) Transwell assay were performed to test the effects of TMZ on the migration and invasion of HERCs induced by HG, respectively. (c) Tubule formation assay and VEGF content were employed to detect the tube-forming ability of HERCs. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. HG. HRECs, Human retinal endothelial cells; VEGF, vascular endothelial growth factor; HG, high glucose.
Figure 3. TMZ decreased the permeability of HRECs cells induced by HG, and increased the expression of Claudin-5 and VE-cadherin proteins.
(a) FITC-albumin was used to detect the permeability of monolayer HRECs. The expression levels of VE-Cadherin (b) and Claudin-5 (c) proteins in HRECs were detected by immunofluorescence. ***P < 0.001 vs. NG, MA, or HG. HRECs, Human retinal endothelial cells; NG, normal glucose; MA, mannitol; HG, high glucose.
Figure 4. TMZ inhibited autophagy mediated by PI3K/Akt/mTOR pathway.
(a) Expression levels of PI3K/Akt/mTOR pathway-related proteins were measured via western blotting. (b) The expression level of Beclin 1 was detected by immunofluorescence. (c) Western blot was conducted to assay the expression of autophagy-related proteins. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. NG, MA, or HG. NG, normal glucose; MA, mannitol; HG, high glucose.
Figure 5. RAPA reversed the inhibitory effect of TMZ on HRECs proliferation, migration, invasion and angiogenesis induced by HG.
Representative images of (a) wound healing analysis, (b) Transwell analysis and (c) tubule formation experiments, and their corresponding quantization levels. *P < 0.05 and ***P < 0.001 vs. NG, MA, or HG. NG, normal glucose; MA, mannitol; HG, high glucose. RAPA, rapamycin; HRECs, Human retinal endothelial cells.
Figure 6. RAPA reversed the inhibitory effect of TMZ on HRECs proliferation, migration, invasion and angiogenesis induced by HG.
FITC-albumin was used to detect the permeability of monolayer HRECs. The expression levels of VE-Cadherin (b) and Claudin-5 (c) proteins in HRECs were detected by immunofluorescence. **P < 0.01 and ***P < 0.001 vs. NG, or HG + TMZ. TMZ, Trimetazidine; HRECs, Human retinal endothelial cells; RAPA, rapamycin; NG, normal glucose; HG, high glucose.
