转化生长因子β1对人脐静脉血管内皮细胞活性的影响  

Effect of transforming grouth factor-β1 on the activation of human umbilical vein endothelial cells

作　　者：张鹏[1] 沈雷[1] 纪亮[1] 姚立杰[1] 谢立平[1] 王岩[1] 张际绯[1] 李静平[1] 

机构地区：[1]齐齐哈尔医学院解剖教研室,161006

出　　处：《中国医药》2013年第1期105-108,共4页China Medicine

基　　金：黑龙江省自然科学基金项目（D200957）;齐齐哈尔市科学技术计划项目（SHFZ-D9011）

摘　　要：目的观察转化生长因子β1（TGF—β1）刺激增生性瘢痕成纤维细胞上清液对人脐静脉血管内皮细胞（HUVEC）增殖和趋化能力的影响。方法以增生性瘢痕成纤维细胞为实验组，正常皮肤为对照组，添加TGF—β1（10μg/L）为TGF—β1组，向TGF—β1组内添加丝氨酸-苏氨酸蛋白激酶（Akt）阻断剂Tricirlbine（5Ixmol／L）为Akt阻断剂组，二者皆未添加的瘢痕成纤维细胞为非干扰组，利用各组细胞上清液（条件培养基）培养HUVEC，使用四甲基偶氮唑盐（MTT）法观察各条件培养基对HUVEC增殖的影响，并利用细胞划痕实验和Transwell细胞迁移实验对HUVEC的趋化情况进行评估。结果对照组吸光度值为0．91±0．34，非干扰组1．75±0．15，TGF—B，组3．26±0．58，Akt阻断剂组1．14±0．81。非干扰组吸光度值与对照组和TGF-β1组比较，差异均有统计学意义（均P〈0．01）；而Akt阻断剂组成纤维细胞增殖吸光度值明显低于TGF—β1组（P〈0．01）。细胞划痕24h后，TGF—β1组促使HUVEC划痕的闭合率较非干扰组高（P〈0．01）；而Akt阻断剂组则明显减弱HUVEC划痕闭合的速度（P〈0．01）。Transwell细胞迁移实验中，TGF—β1组迁移的细胞较非干扰组多[TGF—β1组细胞迁移数目：（1718±15）个，非干扰组：（829±2）个，P〈0．01]；而Akt阻断剂组阳性细胞迁移率又明显降低[Akt阻断剂组：（935±11）个，P〈0．01]。对照组血管内皮生长因子（VEGF）（65．2±0．3）ng／L，非干扰组（87．0±0．5）ng／L，TGF—β1组（132．7±0．4）ng／L，Akt阻断剂组（70．5±0．6）ng／L。非干扰组与对照组、TGF-β1组与非干扰组、Akt阻断剂组与TGF—β1组比较，差异均有统计学意义（均P〈0．01）。结论TGF—β1可以通过刺激增生性瘢痕成纤维细胞旁分泌VEGF等细胞因子对血管内皮细胞增殖、趋化等能力产生影响。Objective To investigate the effect of conditional media collected from transforming growth factor TGF-β1-stimulated fibroblasts on human umbilical vein endothelial cell（HUVEC） proliferation and migration in vitro using human fibroblasts from normal and hyperplastic scaring skin in cell culture. Methods Normal skin or hyperplastic scar fibroblasts were cultured in supplemented serum-free media （SSFM）, and treated with or without TGF-β1 （ 10 μg/L ） in the presence of vehicle or Serine-Threonine protein kinase （ Akt ） inhibitor tricidbine （5 μmol/L）. Conditional media from cultured fibroblasts were collected and used for HUVEC culture. HUVEC proliferation was evaluated by MTT assay, and HUVEC migration was assessed by scratch-wounding and Transwell migration system. Results The optical density value of control group was 0.91 ± 0.34, non interference group was 1.75 ±0. 15, 3.26±0.58 in TGF-β1 group, 1.14 ±0.81 in Akt blockers group. The differences were statistically significant （ P 〈 O. 01 ） ; and the fibroblasts proliferation optical density in Akt blocking agent group was significantly lower than optical density of scar proliferation in TGF-131 group （ P 〈 0.01 ）. Cell scratch after 24 h, prompted HUVEC scratch closure rate in TGF-β1 group were significantly increased than non interference group [ （0.24 ± O. 03 ） cm2 vs （0.09 ± 0.02 ） cm2 , P 〈 0.01 ; and Akt antagonist group decreased significantly HUVEC scratch closing speed [ （0.13 ± 0.01 ） cm2 vs （0.24 ± 0.03 ） cm2, P 〈 0.011. Transwell cell migration assay, migration of cells in TGF-β1 group and non interference group had significant difference （1718± 15 ） vs （829 ± 2）, P 〈 0.01 ） ; and positive cell migration rates were significantly lower in Akt blockers group （935± 11 ） vs （ 1718 ± 15）, P 〈 0. 01 ）. The VEGF in control group, non interference group, TGF-β1 group, Akt antagonist group had significant differences （ all P 〈 0.01 ）. Conclusions The TGF-β1 c

关 键 词：转化生长因子Β1 血管内皮细胞 人脐静脉 

分 类 号：R751[医药卫生—皮肤病学与性病学]