摘要
目的:探讨理肺消积丸通过调控血管生成相关基因表达对肺癌小鼠肿瘤的抑制作用。方法:将30只C57BL/6小鼠采用皮下接种Lewis细胞方式建立肺癌小鼠模型,按照随机数字表法分为模型组、理肺消积组和顺铂组,每组10只。造模次日起依据分组给药,理肺消积组给予理肺消积丸水溶液205 mg/(kg·d)灌胃,等体积生理盐水腹腔注射;顺铂组每隔1 d腹腔注射顺铂0.5 mg/kg,等体积双蒸水灌胃;模型组给予等体积双蒸水灌胃、生理盐水腹腔注射。2周后处死小鼠,留取肿瘤组织,记录小鼠肿瘤质量和体积,苏木精-伊红(HE)染色法观察肿瘤组织病理学变化,免疫组织化学法检测肿瘤组织血管内皮生长因子(VEGF)及微血管密度(MVD),聚合酶链反应阵列(PCR array)筛选血管生成相关差异基因,实时荧光定量聚合酶链式反应(RT-qPCR)检测肿瘤组织中血管生成素样蛋白3(ANGPTL3)、趋化因子配体1(CXCL1)、Ⅳ型胶原α3链(COL4A3)基因表达,蛋白质免疫印迹(WB)法检测肿瘤组织ANGPTL3、CXCL1蛋白表达,免疫荧光法检测肿瘤组织COL4A3蛋白表达。结果:与模型组比较,理肺消积丸组、顺铂组肿瘤组织质量、体积显著减少(P<0.01),肿瘤组织病理形态改善,VEGF蛋白表达显著下调,MVD显著减少(P<0.01);与模型组比较,理肺消积组显著上调或下调的差异表达基因共13个,主要包括ANGPTL3、CXCL1、COL4A3等,参与表皮细胞增殖和血管生成相关调控;与模型组比较,理肺消积组小鼠肿瘤组织ANGPTL3、CXCL1和COL4A3基因与蛋白表达均显著下调(P<0.05)。结论:理肺消积丸可通过降低与血管生成相关基因(VEGF、ANGPTL3、CXCL1)的表达抑制肺癌小鼠肿瘤生长。
Objective:To investigate the inhibitory effect of Lifei Xiaoji Pills on tumor growth in a mouse model of lung cancer by regulating the expression of angiogenesis-related genes.Methods:A lung cancer model was established by subcutaneous inoculation of Lewis cells in 30 C57BL/6 mice,which were randomly divided into three groups according a random number table(n=10 per group):model group,Lifei Xiaoji Pills group,and cisplatin group.From the day following modeling,mice were treated according to group.The Lifei Xiaoji Pills group received drug decoction by gavage at 205 mg/(kg·d)and intraperitoneal injection of normal saline.The cisplatin group received intraperitoneal injections of 0.5 mg/kg cisplatin every other day and gavage with an equal volume of double-distilled water.The model group received gavage with double-distilled water and intraperitoneal saline injection.After 2 weeks,the mice were euthanized and tumors were harvested.Tumor mass and volume were recorded.Histopathological changes in tumor tissues were observed using hematoxylin-eosin(HE)staining.Expression of vascular endothelial growth factor(VEGF)and microvessel density(MVD)were measured by immunohistochemistry.Differentially expressed angiogenesis-related genes were screened using a PCR array.Micro-RNA expression of angiopoietin-like protein 3(ANGPTL3),chemokine ligand 1(CXCL1),and collagen typeⅣalpha 3 chain(COL4A3)was assessed using real-time quantitative PCR(RT-qPCR).Protein expression of ANGPTL3 and CXCL1 was analyzed by Western blot,and COL4A3 expression by immunofluorescence.Results:Compared with the model group,both the Lifei Xiaoji Pills and cisplatin groups exhibited significantly reduced tumor mass and volume(P<0.01),improved tumor histopathology,significantly downregulated VEGF expression,and reduced MVD(P<0.01).Thirteen differentially expressed angiogenesis-related genes were identified in the Lifei Xiaoji Pills group compared with the model group,including ANGPTL3,CXCL1,and COL4A3,which are involved in epithelial cell proliferation and angiogenesis regulation.Expression of ANGPTL3,CXCL1,and COL4A3 at both mRNA and protein levels was significantly downregulated in the Lifei Xiaoji Pills group compared with the model group(P<0.05).Conclusion:Lifei Xiaoji Pills may inhibit tumor growth in lung cancer mice by downregulating angiogenesis-related genes such as VEGF,ANGPTL3,and CXCL1.
作者
谷雨
刘素晓
何志远
丁锦泵
孔德琪
黄慧敏
冯素香
刘学芳
李亚
GU Yu;LIU Suxiao;HE Zhiyuan;DING Jinbeng;KONG Deqi;HUANG Huimin;FENG Suxiang;LIU Xuefang;LI Ya(Department of Respiratory Medicine,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450046,China;Respiratory Pharmacological Laboratory of Chinese Medicine,The First Affiliated Hospital of Henan University Chinese Medicine,Zhengzhou 450000,China;Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Co-construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R.,Zhengzhou 450046,China)
出处
《世界中医药》
北大核心
2025年第10期1672-1678,1685,共8页
World Chinese Medicine
基金
国家自然科学基金项目(82074403)
国家中医临床研究基地科研专项(2022JDZX099)
河南省医学科技攻关计划联合共建项目(LHGJ20230691)
河南省自然科学基金项目(232300420272)
河南省中医药科学研究专项(2022ZY1022)。
