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凌绵聪,樊伟,熊婷,王天磊,潘嘉欣,莫燕丽,刘建浩.清金保肺汤联合针灸辅助治疗非小细胞肺癌的临床及网络药理学研究[J].湖南中医药大学学报英文版,2022,42(4):590-598.[Click to copy
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| 清金保肺汤联合针灸辅助治疗非小细胞肺癌的临床及网络药理学研究 |
| 凌绵聪,樊伟,熊婷,王天磊,潘嘉欣,莫燕丽,刘建浩 |
| (三亚市中医院, 海南 三亚 572000) |
| 摘要: |
| 目的 探讨清金保肺汤联合针灸辅助治疗非小细胞肺癌（non-small cell lung cancer, NSCLC）的临床效果，并借助网络药理学方法建立清金保肺汤多成分-多靶点-多通路的中药复方调控网络与关系网络，结合小鼠实验验证其对于NSCLC的作用机制。方法 选取三亚市中医院2019年1月至2020年12月收治的NSCLC患者80例作为研究对象，分为对照组40例（常规治疗）与观察组40例（在对照组基础上应用清金保肺汤联合针灸治疗），治疗1个月后观察两组治疗效果及评估两组不良反应，监测淋巴细胞亚群百分比及绝对计数的变化情况。通过GEO数据库筛选肺癌肿瘤样本与正常样本的差异基因；运用中药系统药理数据库搜索清金保肺汤药物有效活性成分以及药物作用靶点，运用Cytoscape 3.7.2软件构建成分-靶点基因网络和蛋白质-蛋白质相互作用（PPI）网络；利用R语言进行京都基因与基因组百科全书（KEGG）代谢通路富集分析、基因本体论（GO）富集分析。采用NSCLC细胞株A549右腋下接种建立NSCLC荷瘤模型，蛋白质免疫印迹法对网络药理学筛选的关键作用靶点进行验证。结果 观察组有效率显著高于对照组（P<0.05），不良反应率显著低于对照组（P＜0.05），CD3+T、CD4+T细胞的绝对计数显著上升（P<0.05），B细胞的绝对计数显著下降（P<0.05）。网络药理分析共筛选得到35个靶点基因、59个有效活性成分；PPI网络及拓扑网络分析显示，丝裂原激活蛋白激酶（mitogen-activated protein kinase, MAPK）、EFGR-酪氨酸激酶抑制剂、核因子κB(nuclear factor-κB, NF-κB）等可能是清金保肺汤治疗NSCLC的重要靶点。KEGG通路富集分析显示，清金保肺汤治疗NSCLC主要与酪氨酸激酶受体信号通路、白细胞介素17、MAPK等多条信号通路有关。动物实验结果证实，给药后清金保肺汤低、高剂量组小鼠肿瘤组织中NF-κB和B淋巴细胞瘤-2（B-cell lymphoma-2, Bcl-2）蛋白表达水平均降低（P<0.01），Bcl-2相关X蛋白表达水平升高（P<0.01），验证了网络药理学结果。结论 本研究基于网络药理学并结合小鼠体内实验，初步探究了清金保肺汤治疗肺癌的基因靶向作用以及通路蛋白等信息，为进一步深入研究清金保肺汤对于肺癌的治疗机制奠定了理论基础。 |
| 关键词: 肺癌 清金保肺汤 作用机制 临床效果 网络药理学 信号通路 小鼠 |
| DOI：10.3969/j.issn.1674－070X.2022.04.012 |
| Received:November 09, 2021 |
| 基金项目:海南省卫生健康行业科研项目（19A200155）。 |
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| Clinical and network pharmacology study of Qingjin Baofei Decoction combined with acupuncture in the adjuvant treatment of non-small cell lung cancer |
| LING Miancong,FAN Wei,XIONG Ting,WANG Tianlei,PAN Jiaxin,MO Yanli,LIU Jianhao |
| (Sanya Hospital of Traditional Chinese Medicine, Sanya, Hainan 572000, China) |
| Abstract: |
| Objective To investigate the clinical effect of Qingjin Baofei Decoction combined with acupuncture in the adjuvant treatment of non-small cell lung cancer (NSCLC). By means of network pharmacology, the regulation network and relationship network of Qingjin Baofei Decoction with multi-component, multi-target and multi-pathway were established, and its mechanism of action on NSCLC was studied in vivo in mice. Methods 80 NSCLC patients admitted to Sanya Hospital of Traditional Chinese Medicine from January 2019 to December 2020 were selected as the research objects, and they were divided into control group 40 cases (routine treatment) and observation group 40 cases (on the basis of control group, Qingjin Baofei Decoction combined with acupuncture treatment was applied). After one month of treatment, the therapeutic effect and adverse reactions of the two groups were observed, and the percentage and absolute count of lymphocyte subsets were monitored. GEO database was used to screen the differential genes between lung cancer samples and normal samples. The traditional Chinese medicine systems pharmacology (TCMSP) were used to search the active ingredients and drug targets of Qingjin Baofei Decoction. Cytoscape 3.7.2 software was used to construct the component-target gene network and protein-protein interaction (PPI) network. Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway enrichment analysis and Gene Ontology (GO) enrichment analysis were performed using R language. NSCLC line A549 was inoculated under the right axilla to establish a tumor bearing model. The key targets screened by network pharmacology were verified by Western blot. Results The effective rate of observation group was significantly higher than that of control group (P<0.05), and the adverse reaction rate was significantly lower than that of control group (P<0.05). The absolute count of CD3+T, CD4+T cells were significantly increased (P<0.05), but the absolute count of B cells was significantly decreased (P<0.05). A total of 35 target genes and 59 active ingredients were screened by network pharmacology analysis. PPI network and topological network analysis showed that mitogen-activated protein kinase (MAPK), EGFR tyrosine kinase inhibitor resistance and nuclear factor-κB (NF-κB) may be important targets of Qingjin Baofei Decoction in the treatment of NSCLC. KEGG pathway enrichment analysis showed that the treatment of NSCLC by Qingjin Baofei Decoction was mainly related to tyrosine kinase receptor signaling pathway, interleukin 17, MAPK and other signaling pathways. Animal experimental results confirmed that, after administration, the expression levels of NF-κB and B-cell lymphoma-2 (Bcl-2) in tumor tissues of mice in Qingjin Baofei Decoction low-dose and high-dose groups decreased (P<0.01), the expression level of Bcl-2 related X protein increased (P<0.01), which verified the results of network pharmacology. Conclusion Based on network pharmacology and in vivo experiment in mice, this study preliminarily explores the gene targeting effect and pathway protein of Qingjin Baofei Decoction in the treatment of lung cancer, which lays a theoretical foundation for further verifying the therapeutic mechanism of Qingjin Baofei Decoction for lung cancer. |
| Key words: lung cancer Qingjin Baofei Decoction mechanism of action clinical effect network pharmacology signaling pathway mice |
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