基于网络药理学和体内外实验研究臭壳虫对非小细胞肺癌增殖、迁移和侵袭的影响  

Blaps rynchopetera affects proliferation,migration,and invasion of non-small cell lung cancer:a study based on network pharmacology and in vivo and in vitro experiments

作　　者：李秀育 马科[1,2] 闫靖楠 尤芳程 马璐 LI Xiu-yu;MA Ke;YAN Jing-nan;YOU Fang-cheng;MA Lu(Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Ningxia Ethnic Medicine Modernization,Ministry of Education,Yinchuan 750004,China;Ningxia Hui Autonomous Region Hospital of Traditional Chinese Medicine and Ningxia Chinese Medicine Research Center,Yinchuan 750021,China)

机构地区：[1]宁夏医科大学,宁夏银川750004 [2]宁夏少数民族医药现代化重点实验室,宁夏银川750004 [3]宁夏回族自治区中医医院暨中医研究院,宁夏银川750021

出　　处：《中国中药杂志》2023年第13期3576-3588,共13页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81960860)。

摘　　要：基于网络药理学、分子对接技术和体内外实验研究臭壳虫抗非小细胞肺癌的分子作用机制。文献检索收集臭壳虫成分,通过TCMSP数据库筛选臭壳虫的活性成分,利用PharmMapper数据库获取臭壳虫活性成分的靶点;检索DrugBank、Gene-Cards、OMIM、TTD、PharmGKB数据库中非小细胞肺癌相关的疾病靶点蛋白,再通过绘制韦恩图取臭壳虫活性成分靶点和非小细胞肺癌疾病靶点的交集;通过Cytoscape软件构建“药物成分-靶点”网络和蛋白-蛋白互作(PPI)图,并根据Centiscape插件进行关键靶点的筛选;利用DAVID数据库对上述关键靶点进行GO和KEGG富集分析;采用AutoDock软件对臭壳虫活性成分与其对应的关键靶点进行分子对接验证。结果共获取臭壳虫抗非小细胞肺癌的31种活性成分及72个潜在靶点,11个关键靶点。分子对接结果表明,臭壳虫活性成分与关键靶点之间具有较好的结合活性。制备臭壳虫含药血清,体外培养人肺腺癌A549细胞株,通过CCK-8法检测不同干预时间的空白对照组、不同浓度臭壳虫含药血清组、顺铂组和联合用药组(臭壳虫含药血清+顺铂)对人肺腺癌A549细胞生长的抑制率;通过细胞划痕实验和细胞迁移侵袭实验检测A549细胞迁移和侵袭能力;通过Western blot法检测A549细胞中Bax、Bcl-2、caspase-3、CDC42、SRC、VEGF蛋白表达水平。C57BL/6小鼠皮下接种Lewis肺腺癌细胞,按平均瘤体随机分为模型对照组、臭壳虫组、顺铂组、联合用药组(臭壳虫+顺铂)4组,每组12只;治疗期间隔天监测记录小鼠体质量和肿瘤的长径(a)与短径(b),计算肿瘤体积(0.52ab^(2)),连续用药14 d后处死小鼠,收集肿瘤组织、脾脏及胸腺称重并计算抑瘤率和免疫器官指数;采用HE染色观察各组肿瘤的组织形态,IHC法检测各组肿瘤组织中Bax、Bcl-2、caspase-3、CDC42、SRC、VEGF蛋白阳性表达情况。体内外实验结果表明,臭壳虫组及联合用药组�Network pharmacology,molecular docking,and in vivo and in vitro experiments were employed to study the molecular mechanism of Blaps rynchopetera Fairmaire in the treatment of non-small cell lung cancer(NSCLC).The components of B.rynchopetera were collected by literature review,and the active components were screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).PharmMapper was used to obtain the targets of the active components.The targets of NSCLC were obtained from DrugBank,GeneCards,OMIM,TTD,and PharmGKB.The Venn diagram was drawn to identify the common targets shared by the active components of B.rynchopetera and NSCLC.The "drug component-target" network and protein-protein interaction(PPI) network were constructed by Cytoscape,and the key targets were screened by Centiscape.Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment of the above key targets were performed by DAVID.AutoDock and PyMOL were used for the molecular docking between the key targets and corresponding active components.A total of 31 active components,72 potential targets,and 11 key targets of B.rynchopetera against NSCLC were obtained.The active components of B.rynchopetera had good binding activity with key targets.Further,the serum containing B.rynchopetera was prepared and used to culture human lung adenocarcinoma A549 cells.The CCK-8 assay was employed to determine the inhibition rates on the growth of A549 cells in blank control group and those exposed to different concentrations of B.rynchopetera-containing serum,cisplatin,and drug combination(B.rynchopetera-containing serum+cisplatin) for different time periods.The cell migration and invasion of A549 cells were detected by cell scratch assay and Transwell assay,respectively.Western blot was employed to determine the expression levels of B-cell lymphoma-2(Bcl-2),Bcl-2-associated X(Bax),caspase-3,cell division cycle 42(CDC42),proto-oncogene tyrosine-protein kinase SRC,and vascular endothelial

关 键 词：臭壳虫 非小细胞肺癌 网络药理学 分子对接 体内外实验 

分 类 号：R285[医药卫生—中药学]