本研究旨在通过芥子油诱导的乙酰胆碱表达降低来探究其作为延缓结肠癌进展的潜力。研究设计包括体外细胞实验和体内动物实验，利用芥子油调控乙酰胆碱表达水平并评估其对结肠癌的影响。细胞实验中，将六种浓度(10^-2, 10^-3, 10^-4, 10^-5, 10^-6和10^-7 mol/L)的乙酰胆碱引入结肠癌细胞培养基，以监测细胞增殖。动物实验包括将CT26结肠癌细胞皮下植入28只Balb/c小鼠中，分为实验组和对照组。在肿瘤形成后，两组小鼠均饲以标准饲料两周。从眼部采集血液样本测定血清中的乙酰胆碱浓度。另外，将Balb/c小鼠接种CT26源自结肠癌细胞，并分为实验组和对照组。每组共有14只小鼠，实验组喂食芥子油，对照组喂食豆油。两周后，分别评估两组小鼠的血清乙酰胆碱表达。牺牲后，剖取皮下肿瘤并用游标卡尺测量肿瘤尺寸。培养基中乙酰胆碱浓度增加与细胞增殖逐渐上升呈相关性，达到10^-5 mol/L时显著统计学差异。比较分析显示，带有肿瘤的Balb/c小鼠相对乙酰胆碱表达水平显著升高，与正常Balb/c 小鼠相比。实验组小鼠的血清乙酰胆碱浓度明显低于对照组。芥子油的给药有效地抑制了正常Balb/c小鼠的乙酰胆碱表达，从而延缓了肿瘤生长。这些发现强调了芥子油降低血清乙酰胆碱表达的潜力，从而延缓结肠癌的进展。本研究表明，芥子油通过调控内部环境中的乙酰胆碱表达，具有阻碍结肠癌生长的潜力。

This study aims to investigate the potential of mustard oil-induced reduction in acetylcholine expression as a means to delay the progression of colon cancer within the internal environment.The study design in this research involved both in vitro cellular experiments and in vivo animal experiments to employ mustard oil to modulate acetylcholine expression levels and evaluate its impact on colon cancer. Cellular experiments involved the introduction of six concentrations of acetylcholine (10-2, 10-3, 10-4, 10-5, 10-6, and 10-7 mol/L) into colon cancer cell cultures to monitor cell proliferation. Animal experiments encompassed the subcutaneous CT26 colon cancer cells implantation into 28 Balb/c mice, divided into experimental and control groups. After tumor establishment, both groups were fed standard diets for two weeks. Serum acetylcholine concentrations were measured from eye blood samples. Additionally, Balb/c mice were inoculated with CT26-derived colon cancer cells and further categorized into experimental and control groups. A total of 14 mice comprised each group, with experimental mice fed mustard oil and control mice fed soybean oil. Post two weeks, serum acetylcholine expression in both groups was assessed. After sacrifice, subcutaneous tumors were excised, and tumor dimensions were measured using a Vernier scale.Acetylcholine concentration augmentation in the culture medium corresponded to gradual cell proliferation escalation, peaking at 10-5 mol/L, exhibiting statistical significance. Comparative analysis revealed significantly elevated relative acetylcholine expression levels in Balb/c mice with tumor-bearing colon cancers compared to normal Balb/c mice. Experimental group mice exhibited substantially lower serum acetylcholine concentrations than control group mice. Mustard oil administration effectively curtailed acetylcholine expression in normal Balb/c mice, consequently retarding tumor growth. These findings underscore mustard oil's potential to diminish serum acetylcholine expression, thereby delaying colon cancer progression.This study suggests that mustard oil's modulation of acetylcholine expression within the internal environment holds the potential for impeding colon cancer growth.