基于TLR4/NF-κB/NLRP3通路探讨荆芥连翘汤对溃疡性结肠炎（大肠湿热证）大鼠的作用及机制*

作者：邱爱珠1，徐晔青2，欧阳翌国1，李媛彬1，王紫薇1

单位：1.湖南中医药高等专科学校医学院，湖南 株洲 412000； 2.株洲市中心医院，湖南 株洲 412000

引用：引用：邱爱珠，徐晔青，欧阳翌国，李媛彬，王紫薇.基于TLR4/NF-κB/NLRP3通路探讨荆芥连翘汤对溃疡性结肠炎（大肠湿热证）大鼠的作用及机制[J].中医药导报,2025,31(12):54-60.

DOI：10.13862/j.cn43-1446/r.2025.12.009

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摘要：

目的：观察荆芥连翘汤对溃疡性结肠炎（大肠湿热证）模型大鼠的影响，并基于TLR4/NF-κB/NLRP3信号通路探讨其作用机制。方法：将55只SPF级SD大鼠随机分为5组：正常组、模型组、西药组、灌胃组及灌肠组，每组11只。除正常组外，其余各组建立溃疡性结肠炎（大肠湿热证）大鼠模型。造模成功后，西药组采用美沙拉嗪[0.42 g/（kg·d）]进行灌胃，1次/d；灌胃组、灌肠组分别采用荆芥连翘汤[38.84 g/（kg·d）]进行灌胃、灌肠；正常组、模型组给予0.9%氯化钠注射液灌胃。以上处理持续2周。其间观察大鼠一般行为变化并进行疾病活动指数（DAI）评分。实验结束后，使用HE染色法观察大鼠结肠组织病理变化，免疫组化检测大鼠结肠组织中TLR4、NF-κB、NLRP3、ASC、pro-IL-1β、Caspase-1的蛋白表达情况，实时荧光定量-聚合酶链式反应（qRT-PCR）检测TLR4 mRNA表达情况，酶联免疫吸附试验（ELISA）法检测大鼠血清炎症因子（IL-1β、TNF-α）和抑炎因子（IL-10）的表达情况。结果：与正常组比较，模型组小鼠疾病活动指数DAI评分、血清IL-1β、TNF-α均增加（P<0.05），IL-10下降（P<0.05），结肠组织出现明显的病理学改变，其炎症因子表达含量升高，结肠中TLR4 mRNA及TLR4、NF-κB、NLRP3、Caspase-1、ASC、pro-IL-1β等蛋白表达显著上调（P<0.05）。与模型组比较，各给药组大鼠DAI评分、血清炎症因子IL-1β、TNF-α表达均降低，IL-10升高（P<0.05），结肠中TLR4 mRNA及TLR4、NF-κB、NLRP3、Caspase-1、ASC、pro-IL-1β等蛋白表达显著下调（P<0.05）。与西药组比较，灌胃组大鼠IL-10升高、TLR4 mRNA表达下降、IL-1β降低（P<0.05），其余指标则差异无统计学意义（P>0.05）。灌肠组在降低炎症因子IL-1β、抑制TLR4 mRNA表达方面优于灌胃组（P<0.05）。结论：荆芥连翘汤可抑制TLR4/NF-KB/NLRP3通路激活，降低炎症反应，改善UC（大肠湿热症）大鼠症状和肠黏膜损伤，且灌肠给药相比灌胃给药具有一定优势。

关键词：溃疡性结肠炎；大肠湿热证；荆芥连翘汤；SD大鼠；灌胃；灌肠；TLR4/NF-κB；NLRP3

Abstract：

Objective: To observe the effects of Jingjie Lianqiao decoction on ulcerative colitis (damp-heat syndrome of large intestine) model rats and to explore its mechanism of action based on the toll-like receptor 4 (TLR4)/ nuclear factor kappa B (NF-κB)/nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 (NLRP3) signaling pathway. Methods: Totally 55 SPF-grade SD rats were randomly divided into five groups, including normal group, model group, Western medicine group, gastric administration group, and enema group, with 11 rats in each group. Except for the normal group, the other groups established ulcerative colitis (damp-heat syndrome of large intestin) rat models. After successful modeling, the Western medicine group received meclizine at a dose of 0.42 g/(kg·d) via gastric administration once daily. The gastric administration group and enema group received Jingjie Lianqiao decoction at a dose of 38.84 g/(kg·d) via gastric and enema administration, respectively. The normal group and model group received physiological saline via gastric administration. All treatments were conducted for two weeks. During this period, general behavioral changes of the rats were observed, and disease activity index (DAI) scores were recorded. At the end of the experiment, colon and serum samples were collected, and HE staining was used to observe histopathological changes in the colon tissue. Immunohistochemistry was performed to detect protein expression levels of TLR4, NF-κB, NLRP3, ASC, pro-IL-1β, and Caspase-1 in the colon tissue. qRT-PCR was used to detect TLR4 mRNA expression. Enzyme-linked immunoadsordent assay (ELISA) method was employed to detect the expression levels of inflammatory factors (IL-1β, TNF-α) and anti-inflammatory factors (IL-10) in the mouse blood. Results: Compared to the normal group, the disease activity index DAI score, and serum inflammatory factors (IL-1β and TNF-α) in the model group mice were significantly increased (P<0.05), while IL-10 was significantly decreased (P<0.05). There were significant pathological changes in the colon tissue, with elevated expression of inflammatory factors in model group. The expression levels of TLR4 mRNA, TLR4, NF-κB, NLRP3, Caspase-1, ASC, and pro-IL-1β in the colon were significantly upregulated in model group (P<0.05). Compared to the model group, the DAI scores and serum inflammatory factor (IL-1β and TNF-α) expression in all drug-administered groups were significantly reduced, while IL-10 was significantly increased (P<0.05). The expression levels of TLR4 mRNA, TLR4, NF-κB, NLRP3, Caspase-1, ASC, and pro-IL-1β in the colon were significantly downregulated in all drug-administered groups (P<0.05). Compared with the Western medicine group, the gavage group showed increased IL-10, decreased TLR4 mRNA expression, and reduced IL-1β (P<0.05), while the remaining indicators showed no statistically significant differences (P>0.05).  The enema group performed better than the gavage group in reducing inflammatory factors IL-1β, and inhibiting TLR4 mRNA expression (P<0.05). Conclusion: Jingjie Lianqiao decoction can inhibit the activation of TLR4/NF-KB/NLRP3 pathway, reduce inflammatory response, improve symptoms and intestinal mucosal damage in UC (damp-heat syndrome of alarge intestine) model rats, and enema administration has certain advantages compared with gastric administration.

Key words：ulcerative colitis; damp-heat syndrome of large intestine; Jingjie Lianqiao decoction; SD rats; gavage; enema; TLR4/NF-κB; NLRP3

发布时间：2025-12-31

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