目的 探討經(jīng)方薏苡附子敗醬散（YYFZBJS）對(duì)自發(fā)結(jié)直腸癌的基因工程小鼠（Villin-CreER^T2Kras^G12D/＋Apc^flox/＋）的抗腫瘤作用機(jī)制。方法 Villin-CreER^T2Kras^G12D/＋Apc^flox/＋結(jié)直腸癌小鼠隨機(jī)分為模型組和YYFZBJS低、中、高劑量（3.825、7.650、15.300 g·kg^-1）組，對(duì)照組采用野生C57BL／ 6J小鼠。YYFZBJS低、中、高劑量組ig給予相應(yīng)劑量的YYFZBJS，對(duì)照組和模型組ig給予0.9%氯化鈉溶液12周。檢測小鼠腫瘤形成數(shù)目、腫瘤大小和腫瘤負(fù)荷量。HE染色觀察腫瘤分級(jí)，免疫組織化學(xué)分析表征腫瘤細(xì)胞增殖的蛋白5-溴脫氧尿嘧啶核苷（BrdU）的表達(dá)變化。運(yùn)用中藥系統(tǒng)藥力學(xué)TCMSP平臺(tái)分析YYFZBJS的主要活性成分，并與結(jié)直腸癌靶點(diǎn)取交集，利用STRING數(shù)據(jù)庫和Cytoscape構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)獲得核心靶點(diǎn)；采用Metascape數(shù)據(jù)庫對(duì)交集靶點(diǎn)進(jìn)行基因本體（GO）注釋及京都基因與基因組百科全書（KEGG）通路富集分析，Cytoscape構(gòu)建“藥物-化合物-靶點(diǎn)”網(wǎng)絡(luò)圖，利用AutoDock Tools對(duì)關(guān)鍵活性成分和核心靶點(diǎn)進(jìn)行分子對(duì)接。Western blotting檢測腸道腫瘤組織中吲哚胺-2,3-雙加氧酶1（IDO1）/芳香烴受體（AhR）/β-連環(huán)蛋白（β-catenin）途徑蛋白表達(dá)水平。結(jié)果 與對(duì)照組相比，模型組100%的小鼠結(jié)腸中出現(xiàn)腫瘤，主要為高級(jí)別上皮內(nèi)瘤變。與模型組相比，YYFZBJS低、中、高劑量組小鼠的毛色、生存狀態(tài)均有所改善，腫瘤數(shù)量分別為7.88±2.00（YYFZBJS低劑量組）、6.13±2.17（YYFZBJS中劑量組）和3.13±1.55（YYFZBJS高劑量組），差異具有統(tǒng)計(jì)學(xué)意義（P＜0.01）。相較于模型組小鼠，YYFZBJS低、中、高劑量組的腫瘤組織中BrdU表達(dá)明顯降低（P＜0.05）。網(wǎng)絡(luò)藥理學(xué)結(jié)果顯示YYFZBJS與結(jié)直腸癌共有靶點(diǎn)183個(gè)，包含IDO1、AhR、β-catenin等，涉及癌癥途徑、色氨酸代謝通路等。與模型組小鼠相比，YYFZBJS高劑量組的腸道腫瘤組織的IDO1、AhR和β-catenin蛋白的表達(dá)顯著下調(diào)（P＜0.01）。結(jié)論 YYFZBJS能減緩大腸腫瘤的發(fā)展進(jìn)程，降低腫瘤的惡性分化程度，其作用機(jī)制可能是通過調(diào)節(jié)色氨酸介導(dǎo)的IDO1/AhR/β-catenin途徑有效抑制自發(fā)結(jié)直腸癌的發(fā)生。

Objective To investigate the effect of Yi-Yi-Fu-Zi-Bai-Jiang-San (YYFZBJS) on genetically engineered mice with spontaneous colorectal cancer (Villin-CreER^T2Kras^G12D/＋Apc^flox/＋). Methods Villin-CreER^T2Kras^G12D/＋Apc^flox/＋ colorectal cancer mice were randomly assigned to the model group, YYFZBJS (3.825, 7.650, 15.300 g·kg^-1) dose groups and the wild-type C57BL/6J mice served as the blank control group. The traditional Chinese medicine group was administered YYFZBJS via gavage, while the control and model groups received normal saline gavage for a duration of 12 weeks. Tumor formation, tumor size, and tumor load were subsequently assessed. HE staining was used to observe the tumor grade, and immunohistochemical analysis was performed to characterize the expression of the protein 5-bromo-2'-deoxyuridine (BrdU), which characterizes the proliferation of tumor cells. The principal bioactive constituents of YYFZBJS were identified utilizing the TCMSP analysis platform and subsequently cross-referenced with colorectal cancer-associated targets. Core targets were elucidated through the construction of a protein-protein interaction (PPI) network, employing the STRING database in conjunction with Cytoscape software. Further, gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses for the intersection targets were performed using the Metascape database. Cytoscape was utilized to construct the "drug-compound-target" network diagram, while AutoDock Tools facilitated the molecular docking analysis between key active ingredients and core targets. The expression levels of proteins involved in the IDO1/AhR/β-catenin pathway in intestinal tumor tissues were subsequently assessed using Western blotting. Results In comparison to the normal control group, 100% of the mice in the model group developed colon tumors, predominantly high-grade intraepithelial neoplasia. Relative to the model group, the YYFZBJS intervention group exhibited improvements in hair color and survival status. The tumor counts in the intervention groups were 7.88±2.00 (low dose), 6.13±2.17 (medium dose), and 3.13±1.55 (high dose), respectively, with statistically significant differences (P < 0.01). Additionally, the expression of BrdU in tumor tissues was significantly reduced in the Chinese medicine intervention group compared to the model group (P < 0.05). The network pharmacological analysis revealed that YYFZBJS shares 183 common targets with colorectal cancer, including IDO1, AhR, and β- catenin, which are implicated in both cancer pathways and the tryptophan metabolic pathway. In comparison to the model group, the expression levels of IDO1, AhR, and β-catenin proteins in the intestinal tumor tissues of the YYFZBJS high-dose group were significantly down-regulated (P < 0.01). Conclusion YYFZBJS has been demonstrated to decelerate the progression of colorectal tumors and reduce the degree of malignant differentiation of tumors. The underlying mechanism of action is posited to involve the regulation of the IDO1/AhR/β-catenin pathway, mediated by tryptophan, thereby effectively inhibiting the occurrence of spontaneous colorectal cancer.

R285.5

國家自然科學(xué)基金資助項(xiàng)目（82204887）;上海交大醫(yī)工交叉項(xiàng)目（YG2022QN082）;上海市科委生藥支撐項(xiàng)目（23S21901200）;河南省科技廳科技攻關(guān)項(xiàng)目（242102310493）;河南省科技研發(fā)計(jì)劃聯(lián)合基金項(xiàng)目（242301420116）;河南省中醫(yī)藥科學(xué)研究專項(xiàng)課題（2024ZY2145）