Abstract
Background Diabetic retinopathy (DR) is the most common cause of diabetes-induced microvascular complications and it is the leading cause of blindness in working age worldwide. At the present stage,The main treatment for neovascularization and leakage in DR is anti-VEGF therapy, however, anti-VEGF therapy has its limitation,such as single target and short half-life of anti-VEGF drugs. Therefore, clarifying more therapeutic targets according to the molecular mechanism of neovascularization and leakage is needed, treating the disease by a drug which is multi-target and long-acting. Previous studies have shown that artesunate (ART) can inhibit retinal neovascularization and leakage through multiple targets. This study aimed to clarify the new mechanism of ART inhibiting retinal neovascularization and leakage.
Objective To investigate the new mechanism of retinal neovascularization and leakage inhibited by artesunate (ART) .
Methods Human Umbilical Vein Endothelial Cells (HUVEC) were divided into glucose (G) group, 40mmol/L G+ART(G40+ART)group, mannitol (M) control group, dimethyl sulfoxide (DMSO)control group. The concentration gradient of G group is 5.5mmol/L G (G5.5), 25mmol/L G (G25), 40mmol/L G (G40); The concentration gradient of M control group is 5.5 mmol / L G + 19.5 mmol / L M (M25), 5.5 mmol / L G + 34.5 mmol / L M (M40), The concentration gradient of ART of G40 + ART group is G40 + 10ug /ml ART(10A), G40+20ug/ml ART(20A), G40+40ug/ml ART(40A); the volume of DMSO in the DMSO control group is the same as it is in the 40A group. Western blot,and cell Immunofluorescence technique were used to detect the protein expression of ICAM-1 and MMP-9 in each group.
Results Western blot,and cell Immunofluorescence showed that the protein expression of Intercellular adhesion molecule-1（ICAM-1）and Matrix metalloproteinase-9（MMP-9）in G25 group was higher than that in G5.5 group (P<0.01), and it increased in G40 group compared with G25 group (P<0.01);The protein expression of ICAM-1 and MMP-9 in G25 group was higher than that of M25 group (P<0.01),and it increased in G40 group compared with M40 (P<0.01);the protein expression of ICAM-1 and MMP-9 of G40+ART group was lower than that of G40 group,in which,it was lower in 20A group than that of 10A group (P<0.01), and it was lower in 40A group compared with 20A group (P<0.01). The DMSO control group showed that the protein expression of ICAM-1 and MMP-9 in G40+ART was lower than that of G40+DMSO group (P<0.01).
Conclusion The two targets of ICAM-1 and MMP-9 may act as new therapeutic targets of ART to suppress the retinal neovascularization and leakage in DR,offering assistance for ART used in DR to treat the neovascularization and leakage.