何首乌（PM）是增强免疫力的核心草药。还能解毒、消肿、截疟。其主要成分大黄素（EMD）和2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（二苯乙烯苷，TSG）具有良好的抗癌潜力。本研究旨在探讨两者的协同作用EMD和TSG对CRC的影响及其可能机制。利用网络药理学和生物信息学来识别靶点。采用HPLC分析PM中的有效成分并测定主要成分的含量。 HT-29细胞用于体外实验。采用Cell Counting Kit-8（CCK8）和划痕试验检测PM各化学成分对HT-29细胞增殖和迁移的影响，采用Western Bolt（WB）试验评价EMD和P53 通路上的 TSG。体内实验中，通过测量CRC小鼠模型的肿瘤重量和肿瘤体积来评估EMD和TSG的效果，并通过HE染色进行组织学分析。采用定量逆转录聚合酶链式反应（PCR）检测HSP90、P53、COX2、ROS的表达，检测IL-1β、IL-4、IL-6、IL-10、TGF-β、IFN-γ的表达。通过酶联免疫吸附测定（ELISA）。采用WB和免疫组织化学(IHC)检测P53相关蛋白的表达。网络药理学显示PM与结直肠癌通路密切相关，核心靶点包括STAT3和P53；生物信息学表明P53在CRC的发生发展及预后中发挥重要作用；化学分析显示PM中的没食子酸（GA）、顺式TSG、反式TSG、大黄素葡萄糖苷（EMDG）、大黄素甲醚葡萄糖苷（PHYG）、EMD被鉴定和定量； EMD诱导细胞凋亡，TSG抑制HT-29细胞迁移； EMD和TSG协同缩小CRC小鼠肿瘤体积，上调F4/80表达，降低IL-6和TGF-β含量，促进肿瘤氧化，降低肿瘤中P53和STAT3表达。体外实验表明： TSG 抑制癌细胞迁移和 EMD 诱导细胞凋亡。 EMD 和 TSG 对 CRC 具有协同作用，其可能的机制可能是调节细胞因子的表达并抑制 P53 通路。版权所有 © 2024 Elsevier GmbH。版权所有。

Polygonum multiflorum (PM) is a core herb that enhances immunity. It can also detoxify, reduce swelling, and intercept malaria. Its main components, emodin (EMD) and 2,3,5,4'-Tetrahydroxy stilbene-2-O-β-D-glucoside (stilbene glycoside, TSG), have good anti-cancer potential.The study aims to investigate synergic effects of EMD and TSG on CRC and its possible mechanism.Network pharmacology and bioinformatics were used to identify targets. HPLC was used to analyze the effective ingredients in PM and to determine the content of the main ingredients. HT-29 cells were used for in vitro experiments. Cell Counting Kit-8 (CCK8) and scratch test were used to detect the effects of various chemical components of PM on the proliferation and migration of HT-29 cells, and Western Bolt (WB) test was used to evaluate the effects of EMD and TSG on P53 pathway. In vivo experiments, the effects of EMD and TSG were evaluated by measuring tumor weight and tumor volume in CRC mice model and histological analysis were carried out with HE staining. The expressions of HSP90, P53, COX2, and ROS were detected by quantitative reverse transcription polymerase chain reaction (PCR), and IL-1β, IL-4, IL-6, IL-10, TGF-β and IFN-γ were detected by enzyme linked immunosorbent assay (ELISA). WB and Immunohistochemistry (IHC) were used to detect the expression of P53 related proteins.Network pharmacology showed PM closely related to colorectal cancer pathway and the core targets included STAT3 and P53; bioinformatics indicated P53 played an important role in the development and prognosis of CRC; chemical analysis showed identified and quantified gallic acid (GA), cis-TSG, trans-TSG, Emodin glucoside(EMDG), physcion glucoside (PHYG), EMD in PM; EMD induced apoptosis and TSG inhibited migration of HT-29 cells; EMD and TSG could coordinately shrink tumor size of CRC mice, elevate expressions of F4/80, decrease the content of IL-6 and TGF-β, promote tumor oxidized and reduce expression of P53 and STAT3 in the tumor.In vitro experiments showed that TSG inhibited cancer cell migration and EMD induced apoptosis. EMD and TSG had synergic effects on CRC, whose possible mechanism might be to regulate the expression of cytokines and inhibit P53 pathway.Copyright © 2024 Elsevier GmbH. All rights reserved.