Inhibiting Corneal Neovascularization through DMU-212-induced Apoptosis and Autophagy
Objective To investigate the effects of resveratrol analogue trans-3,4,5,4'-tetram-ethylstilbene(DMU-212)on apoptosis and autophagy during the inhibition of corneal neovascula-tion(CNV).Methods Human umbilical vein endothelial cells(HUVECs)were selected for cul-ture and the cells were randomly divided into six groups,all of which were induced and treated with 10 ng·mL-1VEGF and exposed to 0,10,20,40,80,and 120 μmol·L-1DMU-212,respec-tively,and the proliferation of the cells in each group was detected by the CCK-8 assay.The cell migration rate and angiogenesis ability of the control group(0 μmol·L-1 DMU-212 treatment)and DMU-212 group(40 μmol·L-1DMU-212 treatment)were detected by scratch assay and lu-men formation assay,respectively;the apoptosis and autophagy-related protein expression levels of the 2 groups were detected by protein immunoblotting;the levels of apoptosis and autophagy were detected by TUNEL and MDC,respectively.Results The cell proliferation rate of DMU-212 group was significantly inhibited in a concentration dependent manner compared with the control group(F=11.35,P＜0.001).Compared with the control group,the cell migration rate and lumen-forming ability were significantly inhibited in the DMU-212 group(both P＜0.01);the relative ex-pression levels of cleaved caspase3,cleaved caspase9,Bax,Cyto-c,ATG7,LC3-Ⅱ,and Beclin 1 proteins in the DMU-212 group was significantly higher(P＜0.05 or P＜0.01),and the relative expression levels of P62 and Bc12 proteins was significantly decreased(P＜0.01);the fluorescence intensity of both apoptosis probe and autophagy probe was significantly increased in DMU-212 group(P＜0.05 and P＜0.01).Conclusion DMU-212 can induce apoptosis and increase autophagy levels in HUVECs,thereby inhibiting corneal neovascularization.
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