两种血管内皮生长因子抑制剂对血管内皮细胞功能的抑制作用比较  

Comparison of inhibitory effects of two vascular endothelial growth factor inhibitors on proliferation of vascular endothelial cells

作　　者：王蔚[1] 李学东[1] 刘光旭 何燕 WANG Wei;LI Xue-Dong;LIU Guang-Xu;HE Yan

机构地区：[1]首都医科大学附属北京朝阳医院眼科,北京市100020 [2]首都医科大学公共卫生学院流行病与卫生统计学系,北京市100069

出　　处：《眼科新进展》2019年第10期920-923,共4页Recent Advances in Ophthalmology

基　　金：首都医科大学基础-临床科研合作课题(编号:13JL13)~~

摘　　要：目的对比tenomodulin(TNMD)和avastin对血管内皮生长因子(vascular endothelial growth factor,VEGF)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖和体外血管样结构形成的抑制作用。方法将传至3~6代的HUVEC接种于微孔板中贴壁生长,再置于孵箱中培养6 h,分别加入不同剂量VEGF处理24 h,MTT法测量细胞增殖能力,筛选促进细胞最大增殖作用的VEGF剂量A。将HUVEC分别加入A剂量的VEGF+不同剂量(0.25 mg·L^-1、0.50 mg·L^-1、1.00 mg·L^-1、2.00 mg·L^-1)的TNMD或avastin置于孵箱中培养24 h,MTT法检测TNMD和avastin对VEGF诱导的HUVEC增殖抑制的差异。基质胶体实验检测细胞体外血管形成的形态和长度:实验分为4组,A组为阳性对照;B组添加VEGF;C组添加VEGF和avastin;D组添加VEGF和TNMD,共焦显微镜下观察细胞形态,图像处理分析软件量化分析毛细血管样结构的差异。结果MTT在490 nm波长处测定吸光值(A值)检测细胞增殖,结果显示,相同药物浓度时TNMD的A值均明显低于avastin的A值(均为P<0.01)。基质胶体实验结果显示,D组的毛细血管样结构破坏明显;A组、B组、C组和D组细胞总长度分别为(7422±311)μm、(9369±121)μm、(6499±258)μm、(5292±137)μm,两两相比差异均有统计学意义(均为P<0.05)。结论TNMD比avastin对体外培养的HUVEC增殖具有更强的抑制作用。Objective To explore the inhibitory effect of tenomodulin(TNMD)and avastin on human umbilical vein endothelial cells(UVECs)proliferation and capillary-like morphogenesis in vitro induced by vascular endothelial growth factor(VEGF).Methods HUVECs between passages 3 and 6 were inoculated into microplates for growth,and then placed in an incubator for 6 hours.Different doses of VEGF were added for 24 hours.MTT assay was used to measure cell proliferation and screen out the appropriate VEGF dose(A)for the maximum proliferation effect.A dose of VEGF+different doses(0.25 mg·L^-1,0.50 mg·L^-1,1.00 mg·L^-1,200.00 mg·L^-1)of TNMD or avastin was added into HUVEC in the incubator for 24 hours of culture.MTT assay was used to detect the difference of VEGF-induced inhibition of HUVEC proliferation by TNMD and avastin.The length and morphology of capillary-like structures formed by HUVEC was evaluated by Matrigel tube formation assay.Cells were divided into the positive control(A group),A+VEGF group(B group),B+avastin group(C group),B+TNMD group(D group).Cell morphology was observed under confocal microscopy,and image processing analysis software was used to quantify the differences in capillary-like structures.Results MTT measured the absorbance(A value)at 490 nm to detect cell proliferation.The results showed that the A value of TNMD was significantly lower than that of avastin at the same VEGF concentration(both P<0.01).The results of matrix colloidal experiment showed that the capillary-like structure of D group was destroyed obviously;the total length of cells in A group,B group,C group and D group were(7422±311)μm,(9369±121)μm,(6499±258)μm,(5292±137)μm,and the difference between the two groups was statistically significant(all P<0.05).Conclusion TNMD has a stronger inhibitory effect on angiogenesis in vitro than avastin.

关 键 词：tenomodulin AVASTIN 人脐静脉内皮细胞 细胞增殖 

分 类 号：R77[医药卫生—眼科]