基于网络药理学对[药物名称]治疗结肠炎相关结直肠癌的作用机制分析及小鼠验证
[Analysis of therapeutic mechanism of against colitis-associated colorectal cancer based on network pharmacology and validation in mice].
作者信息
Zhang X, Chen Y, Li Z, Shang J, Yuan Z, Deng W, Luo Y, Han N, Yin P, Yin J
机构信息
School of Traditional Chinese Medicine, Shengyang Pharmaceutical University, Benxi 117004, China.
Interventional Cancer Institute of Chinese Integrative Medicine, Shanghai Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2023 Jul 20;43(7):1051-1062. doi: 10.12122/j.issn.1673-4254.2023.07.01.
OBJECTIVE
To explore the therapeutic mechanism of (LSW) against colitis-associated colorectal cancer (CAC) by network pharmacology.
METHODS
TCMSP, BATMAN-TCM, CNKI, PubMed, Genecards, OMIM, and TTD databases were used to obtain the related targets of LSW and CAC. The common targets of LSW and CAC were obtained using Venny online website. The PPI network was constructed using Cytoscape 3.8.2 to screen the core targets of LSW in the treatment of CAC. GO and KEGG enrichment analysis were conducted using DAVID database. The therapeutic effect of LSW on CAC was evaluated in a C57BL/6J mouse model of AOM/DSS-induced CAC by observing the changes in body weight, disease activity index, colon length, and size and number of the tumor. HE staining and RT-qPCR were used to analyze the effect of LSW on inflammatory mediators. Immunohistochemistry and TUNEL staining were used to evaluate the effect of LSW on the proliferation and apoptosis of AOM/DSS-treated colon tumor cells. Immunohistochemistry and Western blotting were used to detect the effects of LSW on the expression of TLR4 proteins in CAC mice.
RESULTS
Network pharmacology analysis identified 69 common targets of LSW and CAC, and 33 hub targets were screened in the PPI network. KEGG pathway enrichment analysis suggested that the effect of LSW on CAC was mediated by the Toll-like receptor signaling pathway. In the mouse model of AOM/DSS-induced CAC, LSW significantly inhibited colitis-associated tumorigenesis, reduced tumor number and tumor load ( < 0.05), obviously improved histopathological changes in the colon, downregulated the mRNA levels of proinflammatory cytokines, and inhibited the proliferation ( < 0.01) and promoted apoptosis of colon tumor cells ( < 0.001). LSW also significantly decreased TLR4 protein expression in the colon tissue ( < 0.05).
CONCLUSION
LSW can inhibit CAC in mice possibly by regulating the expression of TLR4 to reduce intestinal inflammation, inhibit colon tumor cell proliferation and promote their apoptosis.
目的
通过网络药理学探讨连翘败毒丸（LSW）抗结肠炎相关结直肠癌（CAC）的治疗机制。
方法
利用中药系统药理学数据库与分析平台（TCMSP）、中药系统生物学与药物靶点数据库（BATMAN-TCM）、中国知网（CNKI）、PubMed、基因卡片（Genecards）、在线孟德尔人类遗传数据库（OMIM）和治疗靶点数据库（TTD）获取LSW和CAC的相关靶点。使用Venny在线网站获取LSW和CAC的共同靶点。利用Cytoscape 3.8.2构建蛋白质-蛋白质相互作用（PPI）网络，以筛选LSW治疗CAC的核心靶点。使用DAVID数据库进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。通过观察体重、疾病活动指数、结肠长度以及肿瘤大小和数量的变化，在氧化偶氮甲烷/葡聚糖硫酸钠（AOM/DSS）诱导的C57BL/6J小鼠CAC模型中评估LSW对CAC的治疗效果。采用苏木精-伊红（HE）染色和逆转录-定量聚合酶链反应（RT-qPCR）分析LSW对炎症介质的影响。采用免疫组织化学和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色评估LSW对AOM/DSS处理的结肠肿瘤细胞增殖和凋亡的影响。采用免疫组织化学和蛋白质免疫印迹法检测LSW对CAC小鼠Toll样受体4（TLR4）蛋白表达的影响。
结果
网络药理学分析确定了LSW和CAC的69个共同靶点，并在PPI网络中筛选出33个枢纽靶点。KEGG通路富集分析表明，LSW对CAC的作用是通过Toll样受体信号通路介导的。在AOM/DSS诱导的CAC小鼠模型中，LSW显著抑制结肠炎相关的肿瘤发生，减少肿瘤数量和肿瘤负荷（P<0.05），明显改善结肠组织病理学变化，下调促炎细胞因子的mRNA水平，并抑制结肠肿瘤细胞的增殖（P<0.01），促进其凋亡（P<0.001）。LSW还显著降低结肠组织中TLR4蛋白的表达（P<0.05）。
结论
LSW可能通过调节TLR4的表达来减轻肠道炎症、抑制结肠肿瘤细胞增殖并促进其凋亡，从而抑制小鼠的CAC。
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