风湿祛痛胶囊对VEGF诱导的人脐静脉内皮细胞功能的影响

Effect of Fengshi Qutong Capsule on Human Umbilical Vein Endothelial Cells Induced by VEGF

目的:观察风湿祛痛胶囊对血管内皮细胞生长因子(VEGF)诱导的人脐静脉内皮细胞(HUVEC)的增殖、迁移、黏附、侵袭和管腔形成能力的影响,以及对VEGF受体2(VEGFR2)的干预作用。方法:VEGF体外诱导HUVEC,加入风湿祛痛胶囊低、中、高质量浓度(0. 02,0. 1,0. 5μg·L-1)作用后,分别采用噻唑蓝(MTT)比色法、转移小室(transwell)法、黏附实验、细胞侵袭及管腔形成实验检测HUVEC的增殖活性、迁移、黏附、侵袭及管腔形成能力,蛋白免疫印迹法(Western blot)检测细胞中VEGFR2磷酸化水平和蛋白含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测VEGFR2 mRNA表达水平。结果:与正常组比较,VEGF诱导24,48 h后均能显著升高HUVEC的增殖活性(P <0. 01),诱导24 h能明显升高HUVEC的迁移、黏附、侵袭和管腔形成能力(P <0. 01),显著升高HUVEC中VEGFR2磷酸化及蛋白和mRNA表达水平(P <0. 01);与VEGF组比较,风湿祛痛胶囊低、中、高质量浓度组作用48 h均能明显抑制HUVEC增殖活性(P <0. 05,P <0. 01),作用24 h对VEGF诱导的HUVEC迁移、黏附、侵袭和管腔形成能力有明显抑制作用(P <0. 05),也能下调VEGFR2磷酸化及蛋白和mRNA表达水平(P <0. 05)。结论:风湿祛痛胶囊能降低VEGF诱导的HUVEC细胞增殖、迁移、黏附、侵袭及管腔形成能力,这一作用可能与其抑制VEGFR2的磷酸化、蛋白和mRNA表达水平有关。

Objective: To study the effect of Fengshi Qutong capsule on the migration,adhesion,invasion and tube formation of human synovial cells and the phosphorylation and protein expression of vascular endothelial growth factor receptor 2(VEGFR2). Method: With human umbilical vein endothelial cells(HUVEC)as the research object,low,middle and high-dose Fengshi Qutong capsule(0. 02,0. 1,0. 5 μg · L-1) on HUVEC was determined by methye thiazolye telrazlium(MTT) colorimetric assay for the follow-up experiment. The transwell migration,adhesion and transwell invasion test were used to detect the migration and adhesion of the different concentrations of Fengshi Qutong capsule in HUVEC. The expression of VEGFR2 in HUVEC was detected by Western blot,and Real-time PCR was used to detect the content of VEGFR2 mRNA in cells. Result: Compared with normal group,the proliferation of HUVEC was significantly increased after 24 h and 48 h of VEGF induction(P < 0. 01),the migration,adhesion,invasion and lumen formation of HUVEC were significantly increased within24 h(P < 0. 01),and VEGFR2 phosphorylation protein and mRNA expression levels in HUVEC were significantly elevated in HUVEC(P < 0. 01). Compared with VEGF group,0. 02,0. 1 and 0. 5 μg·L-1 Fengshi Qutong capsule were administered in vitro for 48 h to inhibit HUVEC proliferation activity in a dose-dependent manner(P < 0. 05,P < 0. 01). Within 24 h,VEGF-induced HUVEC migration,adhesion,invasion and lumen formation were significantly inhibited(P < 0. 05),and negatively regulated VEGFR2 phosphorylation protein and gene expression levels(P < 0. 05). Conclusion: Fengshi Qutong capsule can inhibit the migration,adhesion,invasion and tube formation of HUVEC. This effect may be related to the inhibition of phosphorylation,and protein and mRNA expression level of VEGFR2.