Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China.
Shanghai Nanyang Model Private High School, Shanghai, China.
Phytomedicine. 2024 Sep;132:155821. doi: 10.1016/j.phymed.2024.155821. Epub 2024 Jun 23.
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.
何首乌是一种增强免疫力的核心草药，具有解毒、消肿、截疟的功效。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 EMD 和 TSG 对 CRC 的协同作用及其可能的机制。
采用网络药理学和生物信息学方法鉴定靶标，高效液相色谱法分析何首乌中的有效成分并测定主要成分的含量。采用 HT-29 细胞进行体外实验。细胞计数试剂盒-8（CCK8）和划痕试验检测何首乌各种化学成分对 HT-29 细胞增殖和迁移的影响，Western Bolt（WB）试验评价 EMD 和 TSG 对 P53 通路的影响。在 CRC 小鼠模型中，通过测量肿瘤重量和体积评估 EMD 和 TSG 的作用，并进行 HE 染色的组织学分析。采用定量逆转录聚合酶链反应（PCR）检测 HSP90、P53、COX2 和 ROS 的表达，采用酶联免疫吸附试验（ELISA）检测 IL-1β、IL-4、IL-6、IL-10、TGF-β和 IFN-γ的表达。WB 和免疫组化（IHC）检测 P53 相关蛋白的表达。
网络药理学显示，何首乌与结直肠癌途径密切相关，核心靶标包括 STAT3 和 P53；生物信息学表明 P53 在 CRC 的发生发展和预后中起重要作用；化学分析表明，何首乌中鉴定并定量了没食子酸（GA）、顺式 TSG、反式 TSG、EMDG、PHYG 和 EMD；EMD 诱导 HT-29 细胞凋亡，TSG 抑制 HT-29 细胞迁移；EMD 和 TSG 可协同缩小 CRC 小鼠肿瘤体积，提高 F4/80 的表达，降低 IL-6 和 TGF-β的含量，促进肿瘤氧化，降低肿瘤中 P53 和 STAT3 的表达。
体外实验表明，TSG 抑制癌细胞迁移，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通过调节细胞因子的表达来抑制 P53 通路。
