血管抑素体外对兔视网膜微血管内皮细胞增生的抑制  

The Inhibitory Effects of Angiostatin on Microvascular Endothelial Cells from Rabbit Retina

作　　者：孙旭芳[1] 曾水清[2] 王宇哲[3] 李晓青[2] 胡燕华[2] 

机构地区：[1]华中科技大学同济医学院附属同济医院眼科,武汉430030 [2]华中科技大学同济医学院附属协和医院眼科,武汉4300223 [3]华中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2004年第5期573-575,578,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的　观察血管抑素对体外培养的兔视网膜微血管内皮细胞生长的抑制作用及对细胞外信号调节蛋白激酶1(ERK 1)活化的影响。方法　利用L Lysine耦联的Sepharose 4B亲和层析柱从血浆中分离和纯化血管抑素。原代培养兔视网膜微血管内皮细胞 ,MTT法检测不同浓度的血管抑素对其生长的抑制作用 ,Westernblot检测血管抑素对血管内皮细胞生长因子刺激的视网膜微血管内皮细胞ERK 1水平的影响。结果　血管抑素在体外能显著抑制视网膜微血管内皮细胞增生 ,其ED50 约为 16 0 μg/ml。VEGF刺激兔视网膜微血管内皮细胞后 ,ERK 1被迅速激活 ,而血管抑素能使其表达量降低 35 6 %。结论　血管抑素有望成为极有临床价值的防治视网膜新生血管形成的新药。Objective To observe the inhibitory effects of angiostatin on microvascular endothelial cells from rabbit retina and the activities of ERK.Methods Angiostatin was purified with L-lysine Sepharose 4B from human plasma. The primary microvascular endothelial cells from rabbit retina were cultured. The inhibitory effects of angiostatin with different concentrations on the growth of microvascular endothelial cells were measured by using MTT method. The effect of angiostatin on the expression of ERK-1 in retinal microvascular endothelial cells pretreated with VEGF was studied by Western blot. Results Angiostatin could inhibit the growth of microvascular endothelial cells from rabbit retina in vitro with the ED_ 50 being 160 _g/ml. In VEGF treated cells, ERK-1 was rapidly activated and angiostatin could redue the expression of ERK-1 by 35.6%. Conclusion Angiostatin might serve as a valuable drug for preventing retinal neovascularization.

关 键 词：血管抑素 微血管内皮细胞 视网膜 体外 ERK 增生 抑制作用 分离和纯化 表达量 原代培养 

分 类 号：R774[医药卫生—眼科] R730[医药卫生—临床医学]