摘要
目的研究碱性成纤维细胞生长因子(bFGF)对体外培养的人眼脉络膜黑色素细胞表达基质金属蛋白酶-2、9(MMP-2、MMP-9)的影响。方法用Western印迹法从蛋白表达水平定量检测bFGF对脉络膜黑色素细胞表达MMP-2、MMP-9的影响,并用免疫荧光组织化学技术进行定位。结果应用等量总蛋白进行Western印迹法分析,用MMP-2抗体检测发现:正常脉络膜黑色素细胞样本有微弱的信号条带,bFGF处理的样本有清晰的信号条带;用MMP-9抗体检测发现:正常脉络膜黑色素细胞样本未见信号表达,bFGF处理的样本有微弱的信号条带。免疫荧光组织化学显示MMP-2、MMP-9反应产物位于细胞浆,且细胞核周围较密集。结论bFGF可上调体外培养的人眼脉络膜黑色素细胞表达MMP-2、MMP-9。
Objective Previous study has proved that matrix metalloproteinases-2 and -9 （MMP-2, MMP-9） and basic fibroblast growth factor （bFGF） are widely expressed on retinal pigment epithelium cell, extraeellular matrix and vascular endothelium cell of ehoriodal neovaseularization membrane. A investigation of effect of bFGF on the expression of MMP-2 and MMP-9 in cultured human ehoroidal melanoeytes was performed in present work. Methods Isolated human ehoroid from 4 eyes of healthy donors was cultivated in free-serum medium with 20 ng/ml bFGF （bFGF-treated group） and without bFGF （control group）. MMP-2 and MMP-9 were quantitatively analyzed by Western blot and their expressions were localized by immunofluroeseenee staining. Results Western blot study showed a weak signal of MMP-2 at relative molecular weight of 71 000 in ehoroidal melanoeytes in control group and a more intense band in bFGF-treated group. No signal of MMP-9 was detected in ehoroidal melanoeytes of control group,while negligible signal was detected at relative molecular weight of 95 000 in bFGF-treated ehoroidal melanoeytes. A feeble expression of MMP-2 was detected, showing a red fluorescence in cytoplasm, and no MMP-9 were expressed in control group. However,a stronger red fluorescence of MMP-2 and weeker red fluorescence of MMP-9 were found in bFGF-treated melanoeytes. Conclusion bFGF can induce increase of production of MMP-2 and MMP-9 in cultured ehoroidal melanoeytes.
出处
《眼科研究》
CSCD
北大核心
2006年第4期385-388,共4页
Chinese Ophthalmic Research
基金
国家自然科学基金资助(30400229)
