目的 采用网络药理学方法，探讨桃莲绞复方（Tao-lian-jiao Formula，TLF）增强全成分肿瘤细胞疫苗（Vaccine）抗结直肠癌作用的分子机制。方法 观察TLF和全成分肿瘤细胞疫苗单用及联合使用对结肠癌CT26荷瘤小鼠体质量和肿瘤质量的影响，HE染色观察肿瘤组织病理变化。通过中药成分数据库（TCMSP、TCMID），PharmMapper数据库和OMIM数据库分别收集TLF化学成分，成分相关靶点和免疫相关靶点，对TLF调控免疫相关的靶点和通路进行预测分析。通过实时荧光定量PCR、Western blotting对预测结果进行实验验证。结果 与模型组比较，TLF组对肿瘤生长无显著抑制作用，反而表现出促进肿瘤生长的趋势，抑制率-20.1%；Vaccine组对肿瘤生长亦无显著抑制作用（P＞0.05），但有抑制趋势，抑制率19.1%；当TLF联合Vaccine（TLFP<0.05），抑制率达51.4%。通过中药成分数据库共收集到TLF所含的57个化学成分，对应570个靶点，同时收集到免疫相关的168个靶点，比对找到TLF免疫调节的潜在作用靶点4个，分子互作后得到24个关键靶点，信号通路（pathway）和基因本体论（Gene Ontology，GO）生物学过程富集分析结果提示干扰素γ受体1（interferon gamma receptor 1，IFNGR1）及下游的JAK1/JAK2-STAT1可能是TLF发挥免疫调控的关键通路。验证实验结果显示，与Vaccine组比较，TLFP<0.05），与预测结果一致。结论 TLF可增强Vaccine的抗结直肠癌作用，具有作为一种有效的肿瘤免疫治疗佐剂的潜力，其作用机制可能与下调IFNGR1表达，抑制JAK1/JAK2-STAT1信号通路活性有关。

Objective To explore the molecular mechanism of Tao-lian-jiao Formula (TLF, 桃莲绞复方) in enhancing anti-colorectal tumor activity of whole tumor cell vaccine by using network pharmacology. Methods To observe the effects of TLF and whole tumor cell vaccine alone or in combination on the body weight and tumor weight of CT26-bearing mice, and the pathological changes of tumor tissues were detected by H&E staining. The chemical ingredients of TLF, ingredients related targets and immune related targets were collected from traditional Chinese medicine (TCM) databases (TCMSP, TCMID), PharmMapper database and OMIM database, respectively. The targets and pathways related with TLF for regulating immunity were predicted and analyzed. The predicted results were verified by real-time fluorescent quantitative PCR and Western blotting. Results Compared with model group, TLF group had no significant inhibitory effect on tumor growth (P>0.05), while showed a tendency to promote tumor growth, with an inhibition rate of -20.1%; Whole tumor cell vaccine group (Vaccine group) also had no significant inhibitory effect on tumor growth (P>0.05), but there was a inhibition rate of 19.1%; And the combination of TLF and whole tumor cell vaccine (TLF+Vaccine group) could significantly inhibit tumor growth (P<0.05), and the inhibition rate was 51.4%. Through the public databases, 57 ingredients of TLF which corresponding to 570 targets and 168 immune related targets were collected. After comparison, four potential targets of TLF regulating immunity were found, and 24 key targets were obtained after PPI interaction. Pathway signaling and GO biological processes enrichment analysis suggested that interferon gamma receptor 1 (IFNGR1) and its downstream JAK1/JAK2-STAT1 may be the key pathways for TLF to regulate immunity. Verification experiment results showed that the expressions of IFNGR1, phosphorylation of Janus kinase 1 (p-JAK1), phosphorylation of Janus kinase 2 (p-JAK2) and signal transducer and activator of transcription 1 (p-STAT1) in tumor tissue of TLF+Vaccine group were significantly lower than that of vaccine group (P<0.05), which was consistent with the predicted results. Conclusions TLF can enhance the anti-colorectal tumor activity of whole tumor cell vaccine, and has the potential to be an effective adjuvant for tumor immunotherapy. Its mechanism may be related to down-regulating the expression of IFNGR1 and inhibiting the activity of JAK1/JAK2-STAT1 signaling pathway.

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广西中医药民族医药传承创新专项课题（GZLC16-26）；广西自然科学基金项目（2020GXNSFAA238024）；广西区教育厅中青年教师基础能力提升项目（2017KY0320）