The influence of ursolic acid on VEGF, COX-2 and MMP-2 expressed in the mouse retinal ischemic model / 中国医师杂志

ABSTRACT

Objective To investigate the influence of ursolic acid on vascular endothelial growth factor ( VEGF) , cycloxygen-ase-2 (COX-2), and matrix metalloproteinases-2 (MMP-2) expressed in the mouse retinal ischemic model , and to explore the mecha-nisms of anti-angiogenesis.Methods Sixty 7-day clean-class C57BL/6J mice were divided randomly into 6 groups [ n =10 mice (20 eyes) per group]:blank control, model control (PBS), positive control (triamcinolone), and ursolic acid (UA) intervention (low-dose, medium-dose, and high-dose).Mice in the blank control group were raised in air , and mice in other groups in(75%±2%)O2 high-oxygen environment for 5 consecutive days .Mice in the model control group and breastfeeding mice were put back in air environ-ment (21%O2 ) on the 12th day after the new-born mice to induce the generation of retinal neovascularization .When models were suc-cessful, the drug treatments were applied immediately to the corresponding groups , with injection of 3μl of sterile PBS in model control group, 3 μl of 1.5, 3.00 and 6.0 μg UA in UA intervention group, and 3 μl of triamcinolone (1 ml∶40 mg) in positive control group, respectively.All mice were killed after overdose anesthesia on the 17th day.Their eyeballs were made into samples and retinal tissue pathological sections with H-E dying method.The positive expressions of VEGF , COX-2, and MMP-2 were detected with immu-nohistochemical method .The fresh retinal tissue homogenate was prepared to detect the protein expressions of VEGF , COX-2, and MMP-2 in retinal tissue with western blot method ,and mRNA expressions of VEGF , COX-2, and MMP-2 were detected with real-time fluorescent quantitative polymerase chain reaction ( RT-PCR) .Results According to protein and mRNA expressions of VEGF , COX-2,and MMP-2 in retinal tissue among six groups , protein expressions of VEGF , COX-2, and MMP-2 in model group were significantly higher than those in blank group ( P 0.05 ) .Each protein expression in the high UA intervention group was significantly lower than that in the low UA intervention group( P <0.05).Conclusions UA inhibited expressions of VEGF, COX-2, and MMP-2 in retinal ischemia model .UA also played an inhibitory role in the formation of neovascularization , and this role was positively correlated with UA dose .