Comparison of Angiogenic Potential of Human Microvascular Endothelial Cells and Human Umbilical Vein Endothelial Cells

Summary BACKGROUND: Endothelial cells cultured in vitro are commonly used as a model for testing the effects of therapeutic or detrimental agents on endothelium. Cells originating from different vascular beds display, however, a heterogeneity of function and phenotype. Here we compared the production of angiogenic growth factors and the sensitivity to exogenous growth factor stimulation of two popular endothelial cell types. METHODS: Experiments were performed on human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1) incubated in optimal conditions for 24–48 h. RESULTS: The profile of the spontaneously produced growth factors differed significantly between the two different cell lines tested. HUVEC did not produce detectable amounts of VEGF, whereas HMEC-1 released 24.9 pg/ml and this amount was significantly increased in response to IL-1. Instead, HUVEC produced high concentrations of soluble form of VEGF receptor-1 (VEGF-R1), whereas the release of VEGF-R1 from HMEC-1 was 10-fold lower. Small amounts of bFGF were found in media from both cell types, but higher levels were detected in HMEC-1 cultures. In contrast, the secretion of interleukin-8 (IL-8) and matrix metalloproteinase-1 (MMP-1) were 30- to 40-fold higher in HUVEC than in HMEC-1. The cell types differed also in their sensitivity to exogenous growth factors. The basal proliferation of HUVEC was very low but could be effectively stimulated by supplementation with VEGF or bFGF. HMEC-1 proliferated spontaneously and their proliferation rate was not further augmented by growth factors. Similarly, the spontaneous outgrowth of capillaries was negligible in HUVEC but well pronounced in HMEC-1. CONCLUSIONS: Production of angiogenic agents and sensitivity to exogenous growth factors is cell-type dependent. HUVEC, which do not release VEGF, can be easily stimulated with exogenous factors, whereas HMEC-1, which are able to produce VEGF, do not respond well to the additional stimulation. Our study demonstrates that conclusions resulting from in vitro experiments performed on only one type of endothelial cells can be misleading.     

转化生长因子-β、白细胞介素-1和碱性成纤维细胞生长因子对人晶状体上皮细胞的影响

目的:研究体外细胞培养中转化生长因子-β(transforma-tion growth factor beta,TGF-β)、白细胞介素-1(interleukin-1,IL-1)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)对人晶状体上皮细胞(human lens epi-thelial cell,HLEC)的增生作用,对白内障术后晶状体后囊浑浊的预防提供依据。方法:首先进行人晶状体上皮细胞的原代培养,传代培养的晶状体上皮细胞分成各5个实验组,分别加入不同剂量的TGF-β、IL-1,设空白对照组,进行3H-TDR掺入实验,观察TGF-β、IL-1、bFGF对晶状体上皮细胞的作用。结果:TGF-β、IL-1、bFGF对在体外的HLEC增生均有促进作用,并随剂量增加而增强(P<0.05)。结论:TGF-β、IL-1、bFGF在白内障术后的后发性白内障的形成发展过程中起促进作用。     

电损伤后局部应用bFGF促进周围神经再生

目的：通过ｂＦＧＦ局部应用,观察其对神经电损伤后再生的促进作用。方法：随机分为实验组和对照组,每组根据观察时间的长短再分为２ｗｋ组、１ｍｏ组和３ｍｏ组,用６０Ｈｚ１２５Ｖ交流电在梨状肌下缘以远损伤３ｃｍ神经,持续时间３ｓ。伤后第２日起实验组每日注射ｂＦＧＦ５００ＡＵ。对照组注射生理盐水。结果：实验组神经的传导速度、肌张力均较对照组有明显的增加。超微结构改变在各时间点上亦较对照组轻微,再生纤维结构较     

bFGF对神经干细胞分化为星形胶质细胞的影响

 目的 观察碱性成纤维细胞生长因子(bFGF)对大鼠脊髓来源神经干细胞分化形成星形胶质细胞亚型的影响。方法：悬浮培养法培养大鼠脊髓来源的神经干细胞,形成神经球后,以免疫荧光法鉴定神经干细胞。把神经干细胞分为A、B两组,A组加入血清,B组加入血清和bFGF,分化培养7 d后,GFAP抗体标记星形胶质细胞,观察其各亚型的比率和形态特点。结果：神经干细胞分化第7d,A组和B组均形成4种亚型星形胶质细胞,分别为胞体延长型,扁平多角形,星形和双极型。B组中双极型星形胶质细胞的突起长度明显超过A组。结论：bFGF可以显著促进脊髓来源神经干细胞分化成的双极型星形胶质细胞的突起生长。     

微渗透压泵耳蜗灌注bFGF对庆大霉素所致豚鼠耳毒的拮抗作用

目的：应用微渗透压泵将碱性成纤维细胞因子(bFGF)导入耳蜗，观察其对庆大霉素所致活体耳蜗毒性的拮抗作用。方法：豚鼠皮下注射庆大霉素造成耳毒性聋的模型。实验将bFGF以微渗透压泵导入实验组右耳蜗，对照组右耳以微渗透压泵导入等量PBS。用扫描电镜观察两组动物右耳毛细胞的损伤，同时比较其用药前后ABR听阈的改变。结果：对照组右耳的损伤明显重于实验组。结论：采用微渗透压泵耳蜗灌注bFGF对庆大霉素所致豚鼠的耳蜗损伤有拮抗作用。     

薏苡仁注射液对小鼠移植性S180肉瘤血管形成抑制的作用

 目的　探讨薏苡仁注射液对肿瘤血管形成抑制作用。方法　通过建立动物肿瘤模型 ,运用薏苡仁实验治疗 ,检测其抑瘤率和肿瘤内血管密度 ,以及瘤体VEGF和bFGF表达的改变。结果　薏苡仁具有明显的抑制S180肉瘤生长作用。实验组的S180瘤体内微血管密度均明显低于对照组 ;免疫组化显示薏苡仁大、中剂量组均可下调S180瘤体内VEGF、bFGF的表达。结论　薏苡仁注射液对肿瘤血管形成均有明显抑制作用 ,降低VEGF、bFGF的表达可能是其抑制肿瘤血管形成的主要机制之一。     

益肾活血胶囊对同型半胱氨酸血症大鼠生长因子的影响

目的：观察益肾活血胶囊对同型半胱氨酸（Hcy）血症大鼠碱性成纤维细胞生长因子(bFGF),转化生长因子 β1(TGF-β1)的影响。方法：将80只Wistar大鼠随机分为模型组、正常组、大剂量益肾活血胶囊组、小剂量益肾活血胶囊组、西药（叶酸、VitB12、VitB6）组、大癴虫丸组、预防组、空白组共8组,每组10只,除正常组外,其他组大鼠造成高半胱氨酸血症大鼠模型,采用原位杂交法检测各组大鼠主动脉壁bFGF、TGF-β1 mRNA表达,观察大鼠造模前后的变化,以验证各组药物的有效性。结果：与模型组相比益肾活血胶囊能够显著降低高Hcy血症大鼠血管内皮细胞bFGF mRNA（P=0.000）、TGF-β1　mRNA（P=0.032）的表达。结论：益肾活血胶囊能够抑制血管平滑肌细胞和内皮细胞的增生,减缓动脉粥样硬化的进程,为临床治疗血管性疾病提供了依据。     

五叶草莓醇提物对S180荷瘤小鼠VEGF、bFGF影响及形态学观察的实验研究

目的:观察单味五叶草莓醇提物对S180荷瘤小鼠的影响,探讨五叶草莓抗肿瘤效果及可能机制。方法:对S180荷瘤小鼠实体瘤模型予以单味五叶草莓醇提物及CTX治疗,观察抑瘤率,胸腺、脾脏指数及肿瘤细胞中VEGF、bFGF的蛋白表达,并进行疗效分析。结果:CTX组及五叶草莓高、低剂量组的抑瘤率分别为51.69%、45.05%、37.77%,提示环磷酰胺的抑瘤率最高,五叶草莓有一定的抑瘤率;五叶草莓高剂量组与模型组比较脾脏指数明显升高,且具有显著差异(P0.01),中药五叶草莓对胸腺及脾脏有一定的保护作用,对免疫器官影响小于CTX。五叶草莓VEGF、bFGF蛋白在小鼠S180肉瘤组织中呈高表达,环磷酰胺及五叶草莓各剂量组可降低VEGF蛋白表达,其中VEGF与模型组相比有显著差异(P0.01)。结论:五叶草莓醇提物对S180荷瘤小鼠的肿瘤有一定的抑制作用;可促进S180荷瘤小鼠免疫器官胸腺、脾脏的生长;提高S180荷瘤小鼠VEGF、bFGF的水平。