化瘀通络中药单体对VEGF诱导的人脐静脉内皮细胞血管新生和炎症反应的影响

Effects of components in stasis-resolving and collateral-dredging Chinese herbal medicines on angiogenesis and inflammatory response of human umbilical vein endothelial cells induced by VEGF

探讨化瘀通络中药单体野黄芩苷(scutellarin,Scu)、丹皮酚(paeonol,Pae)和羟基红花黄色素A(hydroxy safflower yellow A,HSYA)通过调节血管新生和炎症反应治疗银屑病的机制。取体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)进行实验,设置正常组,模型组,低、中、高3种浓度的Scu、Pae、HSYA组,并以VEGFR酪氨酸激酶抑制剂Ⅱ(VEGFR tyrosine kinase inhibitorⅡ,VRI)作为阳性组。采用CCK-8法检测细胞活力;细胞划痕实验检测细胞迁移能力;小管形成实验检测管腔形成能力;Western blot法检测血管内皮细胞生长因子受体2/蛋白激酶B/细胞外信号调节激酶(VEGFR2/Akt/ERK1/2)信号通路蛋白表达;ELISA法检测炎症细胞因子IFN-γ、IL-1β、IL-6、TNF-α分泌水平。结果显示,与模型组相比,Scu、Pae、HSYA 3种剂量均能显著降低细胞活力、抑制细胞迁移和管腔形成(P<0.05,P<0.01);下调VEGFR2、p-VEGFR2、Akt、p-Akt、ERK1/2、p-ERK1/2蛋白表达,其中Scu和Pae下调VEGFR2表达差异显著(P<0.05,P<0.01),其余组有下降趋势。Scu和Pae显著降低IFN-γ和IL-6的水平(P<0.01),HSYA显著降低IFN-γ、IL-1β和IL-6的水平(P<0.01),Scu、Pae、HSYA对TNF-α水平均无显著影响。研究结果表明Scu、Pae、HSYA可能通过抑制VEGFR2/Akt/ERK1/2信号通路和抑制IFN-γ、IL-1β、IL-6的分泌,从而对银屑病相关的血管异常新生和炎症反应起治疗作用。

The present study investigated the mechanism of components in stasis-resolving and collateral-dredging Chinese herbal medicines,including scutellarin(Scu),paeonol(Pae),and hydroxy safflower yellow A(HSYA),in the treatment of psoriasis by regulating angiogenesis and inflammation.The human umbilical vein endothelial cells(HUVECs) cultured in vitro were divided into a normal group,a model group,a VEGFR tyrosine kinase inhibitor Ⅱ(VRI) group,and Scu,Pae,and HSYA groups with low,me-dium,and high doses.Cell viability was detected by the CCK-8 assay.Cell migration was detected by wound healing assay.Tube formation assay was used to measure the tube formation ability.Western blot was used to detect the protein expression of the VEGFR2/Akt/ERK1/2 signaling pathway.The secretion levels of inflammatory cytokines IFN-γ,IL-1β,IL-6,and TNF-α were detected by ELISA.The results showed that compared with the model group,all the Scu,Pae,and HSYA groups could reduce cell viability,inhibit cell migration and tube formation(P<0.05,P<0.01),and down-regulated the protein expression of VEGFR2,p-VEGFR2,Akt,p-Akt,ERK1/2,and p-ERK1/2.Scu and Pae could down-regulate VEGFR2 expression(P<0.05,P<0.01),while other groups only showed a downward trend.Scu and Pae significantly reduced IFN-γ and IL-6 levels(P<0.01),and HSYA significantly reduced the levels of IFN-γ,IL-1β,and IL-6(P<0.01).Scu,Pae,and HSYA had no significant effect on TNF-α.The results suggested that Scu,Pae,and HSYA may exert a therapeutic role in psoriasis-related angiogenesis and inflammation by inhibiting VEGFR2/Akt/ERK1/2 signaling pathway and inhibiting the secretion of IFN-γ,IL-1β,and IL-6.