大黄素抑制VEGF165诱导下人脐静脉内皮细胞的增殖、迁移及侵袭的研究  

Study on the Inhibition of Emodin on the Proliferation,Migration and Invasion of Human Umbilical Vein Endothelial Cells Induced by VEGF165

作　　者：徐卫星 孔凡逍 Xu Weixing;Kong Fanxiao(The Third Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000 China)

机构地区：[1]锦州医科大学附属第三医院,辽宁锦州121000

出　　处：《锦州医科大学学报》2023年第4期33-38,共6页Journal of Jinzhou Medical University

摘　　要：目的观察大黄素(emodin,EMO)对重组人血管内皮生长因子165(recombinant human vascular endothelial growth factor165,VEGF165)诱导下人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖、迁移及侵袭的影响并探讨其机制。方法离体培养HUVECs,MTT检测细胞活力计算EMO最佳作用浓度。实验分组和干预措施如下:对照组(完全培养基培养)、VEGF模型组(VEGF16520 ng/mL预处理72 h)、EMO治疗组(VEGF16520 ng/mL+EMO预处理72 h)。采用划痕实验、Transwell小室垂直迁移、侵袭实验检测各组细胞水平迁移、垂直迁移和侵袭能力,采用细胞免疫荧光实验检测VEGF表达,Western Blot检测VEGF、VEGFR2、ERK和SRC蛋白表达的变化。结果HUVECs活力值(A值)随着EMO浓度增加而降低,半抑制浓度(IC50)为20.27μmol/L,将20μmol/L作为EMO的最佳有效浓度进行后续实验。EMO治疗组相对水平迁移率、垂直迁移率和侵袭率均较VEGF模型组降低,差异均有统计学意义(P<0.05)。EMO治疗组细胞免疫荧光VEGF蛋白表达荧光强度低于VEGF模型组。EMO治疗组VEGF、VEGFR2、ERK和SRC蛋白表达较VEGF模型组降低,差异均有统计学意义(P<0.05)。结论EMO可以有效抑制VEGF165诱导下人脐静脉内皮细胞的增殖、迁移及侵袭能力,可能是通过抑制VEGF/VEGFR2/MAPK/ERK和VEGF/VEGFR2/SRC信号通路实现的。Objective To observe the effect of emodin(EMO)on the proliferation,migration and invasion of human umbilical vein endothelial cells(HUVECs)induced by Recombinant Human Vascular Endothelial Growth Factor165(VEGF165)and explore its mechanism.Methods HUVECs were cultured in vitro and the optimal concentration of EMO was calculated by MTT assay.The experimental grouping and interventions were as follows:control group(complete medium culture),VEGF model group(VEGF16520 ng/mL pre-treated for 72 h),and EMO treatment group(VEGF16520 ng/mL+EMO pre-treated for 72 h).Scratch test,Transwell chamber vertical migration and invasion assay were used to detect the horizontal migration,vertical migration and invasion ability of each group of cells.Immunofluorescence assay was used to detect the expression of VEGF and Western Blot was used to detect the changes of protein expression of VEGF,VEGFR2,ERK and SRC.Results The viability of HUVECs(A value)decreased with the increase of EMO concentration.The semi-inhibitory concentration(IC50)was calculated as 20.27μmol/L,and 20μmol/L was used as the best effective concentration of EMO for subsequent experiments.The relative horizontal migration rate,vertical migration rate and invasion rate of the EMO treatment group were lower than those of the VEGF model group,and the differences were statistically significant(P<0.05).The immunofluorescence intensity of VEGF protein expression in EMO treatment group was lower than that in VEGF model group.The protein expressions of VEGF,VEGFR2,ERK and SRC in EMO treatment group were lower than those in VEGF model group,and the differences were statistically significant(P<0.05).Conclusion EMO can effectively inhibit the proliferation,migration and invasion of human umbilical vein endothelial cells induced by VEGF165,which may be achieved by inhibiting VEGF/VEGFR2/MAPK/ERK and VEGF/VEGFR2/SRC signaling pathways.

关 键 词：大黄素 age-related macular degeneration VEGF HUVEC 新生血管 

分 类 号：R285[医药卫生—中药学]