左金丸通过抑制ERK、p38和AKT信号通路抑制肿瘤血管生成和结肠癌生长  

ZuoJin Wan inhibits tumor angiogenesis and colorectal cancer growth by suppressing ERK,p38 and AKT signaling pathways

作　　者：袁杰[1] 孟娇 闫化茹 常彬 孙明瑜[1] 梁婷玉[1] 王子元[1] YUAN Jie;MENG Jiao;YAN Huaru;CHANG Bin;SUN Mingyu;LIANG Tingyu;WANG Ziyuan(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海201203

出　　处：《海南医科大学学报》2025年第5期343-350,共8页Journal of Hainan Medical University

基　　金：国家自然科学基金资助项目(82374188、82303207);曙光医院四明青年基金(SGKJ-202115)。

摘　　要：目的:研究左金丸通过抑制ERK1/2、p38MAPK和AKT信号通路对肿瘤血管生成的影响。方法:使用不同浓度的左金丸处理含有或不含VEGF的HUVEC和Lovo细胞,利用CCK-8法评估左金丸对HUVEC和Lovo细胞增殖的影响;利用流式细胞术分析左金丸对HUVEC细胞凋亡的影响;采用划痕实验、Transwell实验、管腔形成实验检测左金丸对HUVEC细胞迁移、侵袭和管腔形成的影响;建立人结肠癌Lovo细胞裸鼠皮下移植瘤模型,随机分为5组:对照组(0.9%氯化钠溶液组),左金丸低、中、高剂量组(1 027.5 mg/kg,2 055 mg/kg,4 110 mg/kg)和化疗药5-Fu组(50 mg/kg),每隔1日给药1次。给药第21天处死裸鼠,比较各组肿瘤体积和重量。利用免疫组化法检测各组瘤体中CD34表达(微血管密度)。采用Western blot法检测左金丸对VEGFR2及其下游信号通路的调控作用。结果:左金丸能显著抑制HUVEC细胞和Lovo细胞的增殖,尤其对VEGF诱导的HUVEC细胞增殖的抑制效果更为明显(P<0.05)。左金丸能促进VEGF诱导的HUVEC细胞凋亡、抑制其迁移、侵袭和管腔形成能力(P<0.05)。在裸鼠移植瘤模型中,左金丸剂量依赖性地抑制了移植瘤的体积和重量(P<0.05),并使瘤体微血管密度显著减低(P<0.05)。此外,左金丸还通过抑制VEGF诱导的HUVEC细胞中ERK1/2、p38MAPK和AKT的磷酸化水平发挥作用,表现出剂量依赖性和时间依赖性,但未对VEGFR2磷酸化水平表达产生影响。结论:左金丸能有效抑制肿瘤血管生成,其机制可能与其抑制ERK1/2、p38MAPK和AKT信号通路有关。Objective:To investigate the inhibitory effect of Zuojin Wan(ZJW)on tumor angiogenesis by suppressing the ERK1/2,p38MAPK,and AKT signaling pathways.Methods:HUVEC and Lovo cells were treated with ZJW at various concentrations with or without VEGF.The CCK-8 assay was employed to assess the effect of ZJW on the proliferation of HUVEC and Lovo cells.Flow cytometry was used to observe the effect of different concentrations of ZJW on apoptosis in HUVEC cells.Migaration assay,Transwell assay,and tube formation assay were conducted to evaluate the migration,invasion,and tube-forming ability of HUVECs treated with ZJW.A nude mouse subcutaneous xenograft model of human colon cancer Lovo cells was established,divided into 5 groups:the control group(0.9%saline),low,medium,and high-dose ZJW groups(1027.5 mg/kg,2055mg/kg,4110 mg/kg),and chemotherapy drug 5-Fu group(50 mg/kg),all administered every other day.On the 21st day of treatment,mice were euthanized,and tumor volumes and weights were compared among groups.Immunohistochemistry was used to detect CD34 expression(microvessel density)in tumor tissues of each group.Western blot analysis was performed to investigate the regulatory effects of ZJW on VEGFR2 and its downstream signaling pathways.Results:ZJW inhibited the proliferation of HUVEC and Lovo cells,with a more pronounced inhibitory effect on VEGF-induced HUVEC cell proliferation(P<0.05).ZJW inhibited cell migration,invasion,and tube formation(P<0.05).ZJW suppressed the volume and weight of xenograft tumors in a dose-dependent manner(P<0.05).Compared to the control group,microvessel density in the low,medium,and high-dose ZJW groups showed a dose-dependent decrease(P<0.05).ZJW inhibited the phosphorylation levels of ERK1/2,p38MAPK,and AKT in VEGF-induced HUVEC cells in a dose-and time-dependent manner but had no effect on VEGFR2 phosphorylation expression.Conclusion:ZJW could inhibit tumor angiogenesis,and its mechanism may be associated with the inhibition of the ERK1/2,p38MAPK,and AKT signaling pathways.

关 键 词：左金丸 肿瘤血管生成 结肠癌 ERK1/2 AKT p38MAPK 

分 类 号：R285[医药卫生—中药学]