柚皮铜配合物对激光诱发大鼠脉络膜新生血管形成的抑制作用

Inhibitory effects of naringenin and its complex of copper(Ⅱ) with free ligand on formation of choroidal neovascularization in rats induced by laser

目的:观察柚皮铜配合物对激光诱发大鼠脉络膜新生血管形成的抑制作用。方法:选取雄性Brown-Norway大鼠24只,其中18只建立大鼠脉络膜新生血管模型,6只12眼作正常对照。动物模型随机分为模型对照组和2个不同给药实验组,每组6只。模型建立后,实验组大鼠腹腔注射同等剂量的柚皮素、柚皮铜配合物[20mg/(kg·d)],对照组不给药。于造模后30d取各组视网膜-脉络膜-巩膜复合体标本,在荧光显微镜下观察脉络膜新生血管的面积,采集各组大鼠的脉络膜标本,采用RT-PCR及Western blot方法检测各组大鼠脉络膜组织中VEGF、COX-2、PI3K、MAPK、MMP-2、MMP-9mRNA与蛋白的表达。结果:与模型对照组相比,各实验组大鼠新生血管面积均减少,但柚皮铜配合物组较柚皮素组更明显,差异均具有显著统计学意义(P<0.01);模型组基因和蛋白表达水平均高于正常对照组,差异均有统计学意义(P<0.01);各实验组基因及蛋白表达水平与模型组相比均有所下降,除PI3K,MMP-9外,柚皮铜配合物组下降幅度大于柚皮素组,差异均有统计学意义(P<0.05)。结论:相较于柚皮素,柚皮铜配合物有更好的生物活性,对激光诱发大鼠脉络膜新生血管的形成有更显著的抑制作用。

AIM： To observe the inhibitory effects of formation of choroidal neovascularization （ CNV ） in rats induced by laser by naringenin and complex of copper （ II） with free ligand. METHODS： A total of 24 male Brown-Norway rats were selected including 18 rats as CNV model and 6 as normal comparison. A total of 18 model rats were divided randomly into control, naringenin and its complexes of copper group with 6 rats in each group. Naringenin and its complexes [20mg/（kg· d）] were given once-daily through intraperitoneal （ i.p.） injection after laser treatment for 30d in two experimental groups, respectively. Complex samples of retina, choroid membrane and sclera in each group were taken after 30d of the establishment of animal model.The sizes of CNV lesions were observed under fluorescence microscope. Samples of choroid in each group were taken. The expression of protein and mRNA of COX-2, VEGF, PI3K, MAPK, MMP-2 and MMP-9 were examined by Western blotting and RT-PCR method at 30d. RESULTS：The animals treated with naringenin and its complexes of copper showed significant decrease in their average CNV sizes, compared with that of the vehicle-treated animals （P〈0.01）, especially with naringenin＇s and its complexes of copper group.Compared with the vehicle -treated animals, the mRNA and protein expressions of VEGF, COX -2 and so on were significantly higher than that in the model control group （P〈0.01）.The expression of gene and protein in each experimental group both decreased when compared with model group. Except PI3K, MMP -9, the reduction degree in the naringenin and its complexes of copper group were larger than that in the single naringenin group （ P〈0.05 ）.The differences of reduction degree were statistically significant. CONCLUSION： These results indicate that the biological activity of naringenin can be enhanced by the formation of its complexes, and that complexes of naringenin have stronger inhibition effect on the formation of CNV in rats induced by laser.