Inhibitory Effect Of Ginsenoside RG3 On Choroidal Neovascularization

PurposeThis study aimed to investigate the inhibitory effect of ginsenoside RG3 on choroidal neovascularization in vitro and in vivo and explore its possible mechanism.MethodsIn vitro: The cells were added with DMSO control solution(2?g·ml-1)and ginsenoside RG3 solution(80.0?g · ml-1),respectively,and cell proliferation,migration,and tubule formation were detected by MTT assay,cell scratch assay,and tubule formation assay.Apoptosis was detected by TUNEL staining and flow cytometry,and the expression of VEGF,p-Akt,and p-Erk was detected by Western blot and immunofluorescence staining.In vivo: Twenty male and 20 male C57BL/6J mice were randomly divided into a control group and a ginsenoside RG3 group.One week of subcutaneous injection pretreatment was used to establish a choroidal neovascularization model induced by a semiconductor laser.The fluorescein sodium fundus angiography was performed at 7 days,14 days,21 days,and 28 days after photocoagulation to observe the formation and leakage of CNV.Western blot was used to detect VEGF,p-Akt,p-in the mouse eyeball after 3 days of modeling.Erk and other protein expression.ResultsIn vitro: 1.MTT results showed that HUVECs were treated with ginsenosideRG3.After 12 hours,the absorbance of ginsenoside RG3 group was lower than that of DMSO control group.The difference was statistically significant(P<0.01).Ginsenoside RG3 inhibited the proliferation of HUVECs.2.The cell scratch test results show that cells in the DMSO control group migrated to the scratch-free cell area without growth,whereas the cells of the ginsenoside RG3 group had no significant migration and growth,and the difference was statistically significant(P<0.05).),It was proved that ginsenoside RG3 could inhibit the migration of endothelium;3.Tubule formation test results showed that the number of tubule formation in DMSO control group was higher than that of ginsenoside RG3 group,and the difference was statistically significant(P<0.01).It was confirmed that ginsenoside RG3 The formation of endothelial cell tubules was inhibited.4.TUNEL staining showed that the positive expression of HUVECs in ginsenoside RG3 group was higher than that in DMSO control group.Flow cytometry results also showed that the apoptosis rate of HUVECs in ginsenoside RG3 group was higher than that in DMSO control group;Immunofluorescence staining and Western blot analysis of apoptotic protein Cleaved caspas-3 showed that the relative amount of Cleaved caspas-3 protein in ginsenoside RG3 group The amount is higher than the DMSO control group,indicating that ginsenoside RG3 can induce apoptosis of endothelial cells by regulating the expression of Cleaved caspas-3 protein;5.Immunofluorescence staining and Western blot results show that ginsenoside RG3 group compared with DMSO control group,the relative expression levels of VEGF,p-Akt and p-Erk were all decreased,and the differences were statistically significant(P<0.01).It was proved that ginsenoside RG3 inhibited the expression of VEGF,p-Akt and p-Erk.In vivo: 1.Fundus fluorescein sodium angiography showed that both groups had leakage of fluorescein sodium and formation of choroidal neovascularization.Over time,the leakage of fluorescein sodium was reduced in both groups.However,at the same time point,the fluorescence leakage areaand intensity of ginsenoside RG3 group were lower than that of DMSO control group,which proved that ginsenoside RG3 could inhibit the formation and leakage of choroidal neovascularization;2.Western blotting results showed that laser modeling after 3 days,the relative expression of VEGF,p-Akt,and p-Erk in ginsenoside RG3 group was lower than that in blank group and DMSO control group,and the difference was statistically significant(P<0.01).It was confirmed that ginsenoside RG3 inhibited the expression of VEGF,p-Akt,and p-Erk proteins in mice.Conclusions1.Ginsenoside RG3 inhibits the proliferation and migration of HUVECs by reducing the expression of VEGF,p-Akt,p-Erk and other proteins,and induces the apoptosis of HUVECs.2.Ginsenoside RG3 can inhibit the formation of choroidal neovascularization and reduce the permeability of choroidal capillaries.3.Ginsenoside RG3 can be used as a potential effective drug to provide new ideas for the treatment of wet AMD.