黄芪总皂苷与莪术醇抑制肿瘤血管生成及其对EGFR/PI3K/AKT和HIF-1α/VEGF信号通路的影响  

Astragalosides and Curcumol Inhibit Tumor Angiogenesisand Affect EGFR/PI3K/AKTand HIF-1α/VEGF Signaling Pathways

作　　者：杨倩宇 闫梓乔 李潇 许成勇[3] 窦永起 YANG Qian-yu;YAN Zi-qiao;LI Xiao;XU Cheng-yong;DOU Yong-qi(Hebei University of Chinese Medicine,Shijiazhuang Hebei 050000;ChinesePLA Medical School,Beijing 100853;Hainan Hospital of Chinese PLA General Hospital,Sanya Hainan 572013;Department of Traditional Chinese Medicine,Chinese PLA General Hospital,Beijing 100048)

机构地区：[1]河北中医学院,河北石家庄050000 [2]解放军医学院,北京100853 [3]解放军总医院海南医院,海南三亚572013 [4]解放军总医院中医医学部,北京100048

出　　处：《世界中西医结合杂志》2022年第6期1115-1120,1125,共7页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金项目(81673810)。

摘　　要：目的通过观察黄芪、莪术主要有效抗癌成分黄芪总皂苷、莪术醇单独及联合应用体外抑制肿瘤血管生成的作用及其对EGFR/PI3K/AKT和HIF-1α/VEGF信号通路的影响,进一步阐明临床常用配伍黄芪-莪术的抗癌作用机制和内在物质基础。方法采用CCK8法筛选黄芪总皂苷及莪术醇体外有效浓度,设对照组、黄芪总皂苷组、莪术醇组、两药联合组进行HUVECs细胞体外培养,CCK-8法检测各组细胞增殖抑制率,光镜下观察HUVECs细胞体外成血管能力,Westernblot及RT-qPCR法检测EGFR/PI3K/AKT及HIF-1α/VEGF信号通路相关蛋白及其mRNA的表达。结果干预24 h后,50μmol/L黄芪总皂苷及50μmol/L莪术醇对HUVECs细胞具有较强抑制作用;细胞增殖抑制率分别为黄芪总皂苷组3.06%、莪术醇组3.14%及两药联合组11.75%,两药联合组显著优于单药组(P<0.01);体外成血管能力,黄芪总皂苷组、莪术醇组与对照组比较未显示明显差异(P>0.05),两药联合组的血管管腔数显著少于对照组、黄芪总皂苷组及莪术醇组(P<0.01);各蛋白的表达,黄芪总皂苷组EGFR、PI3K、HIF-1α、VEGF蛋白表达下降,其中EGFR下降尤为显著(P<0.01),莪术醇组HIF-1α、VEGF蛋白表达下降明显(P<0.01),两药联合组EGFR、PI3K、AKT、HIF-1α、VEGF蛋白表达下降,其中EGFR、PI3K、HIF-1α、VEGF降低尤为显著(P<0.01);各mRNA转录,黄芪总皂苷组EGFR、PI3K、AKT、VEGF mRNA表达降低,莪术醇组AKT、HIF-1α、VEGF mRNA表达降低,两药联合组EGFR、PI3K、AKT、HIF-1α、VEGF mRNA表达降低,其中EGFR、PI3K、AKT、VEGF mRNA降低尤为显著(P<0.01)。结论黄芪总皂苷和莪术醇可通过抑制EGFR/PI3K/AKT信号通路、HIF-1α/VEGF信号通路EGFR、PI3K、AKT、HIF-1α、VEGF等相关mRNA转录及蛋白表达,进而抑制血管内皮细胞增殖及毛细血管样结构形成,两者联合作用更强,揭示了黄芪-莪术配伍抑制肿瘤血管生成的主要作用靶点及物质基础。Objective To investigate the effects of single and combined application of astragalosides and curcumol,the primary active anti-cancer components of Astragali Radix and Curcumae Rhizoma on the in vitro inhibition of tumor angiogenesis and the epidermal growth factor receptor(EGFR)/phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(AKT)and hypoxia-induciblefactor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathways and further clarify the anti-cancer mechanism and material basis of the commonly used drug pair,Astragali Radix and Curcumae Rhizoma.Methods The CCK8 method was used to screen the effective concentrations of astragalosides and curcumol.Human umbilical vein endothelial cells(HUVECs)were cultured in vitro and divided into a control group,an astra-galosides group,a curcumol group,and a combination group.The proliferation inhibitory rate of cells in each group was detected by the CCK-8 method,and the angiogenesis ability of HUVECs in vitro was observed under a light microscope.Western blot and RT-qPCR were used to detect the expression of proteins and genes related to the EGFR/PI3 K/AKT and HIF-1α/VEGF signaling pathways.Results After 24 hours of intervention,50μmol/L astragalosides and 50μmol/Lcurcumol showed strong inhibitory effects on HUVECs.The proliferation inhibitory rates of the astragalosides group,the curcumol group,and the combination groups were 3.06%,3.14%,and 11.75%,respectively,suggesting that the combination group was superior to the groups with single application(P<0.01).In terms of the angiogenesis ability,the astragalosides group andthe curcumol group showed no significant difference as compared with the control group(P>0.05),and the number of vascular lumens in the combination group was less than those in the control group,the astragalosides group,andthe curcumol group(P<0.01).In terms of protein expression,the expression of EGFR,PI3 K,HIF-1α,and VEGF in the astragalosides group decreased,especially EGFR(P<0.01),and the expression of HIF-1αand VEGF in the curcum

关 键 词：黄芪总皂苷 莪术醇 血管生成 表皮生长因子受体/磷脂酰肌醇3激酶/蛋白激酶B 缺氧诱导因子-1α/血管内皮生长因子 细胞实验 中药物质基础 

分 类 号：R285.5[医药卫生—中药学]