基于NF-κB-IL-6通路探究开心散致小鼠肠道炎症的分子机制

doi:10.19803/j.1672-8629.20260134

摘要/Abstract

摘要： 目的探究经典名方开心散致小鼠肠道炎症的原因,初步阐释其潜在分子机制,为开心散的临床用药安全提供参考。方法将C57BL/6J小鼠随机分为对照组和（0.5、1.0、2.0、4.0 g·kg^-1）开心散剂量组,通过肠推进试验计算小肠推进率。对给药3 d和8 d的小鼠（对照组和0.5、1.0、1.5 g·kg^-1开心散剂量组）进行下述指标的检测：全自动血液分析仪检测血液学指标,苏木精-伊红（HE）染色观察肠道组织的病理损伤情况,免疫荧光染色检测黏蛋白MUC2的表达及分布情况,Western blot技术分析肠道组织中NF-κB p65、P-IκB、NLRP3、NLRC4、ASC、pro-Caspase1的蛋白表达量,高通量检测技术（Luminex）分析给药8 d后小鼠小肠组织中IL-1β、TNF-α、IL-6、IL-22的细胞因子水平,ELISA分析IL-6因子水平。结果小鼠灌服不同剂量的开心散3 d后,小肠推进率显著降低且呈剂量依赖性。给药8 d空肠、回肠出现病理损伤,包括肠绒毛萎缩、黏膜结构紊乱和肠腔扩张;随着开心散剂量升高,MUC2荧光信号减弱;1.5 g·kg^-1开心散诱导小肠中pro-Caspase1、NF-κBp65、P-IκB的蛋白表达量明显升高,结肠未见明显变化。开心散给药3 d,小肠、结肠组织中上述指标均无显著变化;Luminex与ELISA结果显示1.5 g·kg^-1剂量组IL-6水平显著升高。结论小鼠灌胃给予0.5 g·kg^-1开心散3 d即可引发胀气,并诱发肠道动力障碍,激活NF-κB通路调控IL-6转录引发下游炎症反应,主要靶器官在小肠（空肠、回肠）,具有部位特异性。

关键词: 开心散, 肠道炎症, NF-κB, IL-6, 小肠动力, 黏液屏障, 小鼠

Abstract: Objective To investigate the molecular mechanism through which Kai-Xin-San induces intestinal inflammation in mice in order to ensure its long-term clinical safety. Methods C57BL/6J mice were randomly assigned to the control group and the groups treated with doses of 0.5, 1.0, 2.0 and 4.0 g·kg^-1 before the small intestinal transit rate was determined via an intestinal propulsion test. After 3 or 8 days of administration (The control group and the groups treated with doses of 0.5, 1.0, and 1.5 g·kg^-1), hematological parameters were measured using an automated hematology analyzer. Histopathological injury was examined by hematoxylin and eosin (HE) staining. The expression and distribution of MUC2 were evaluated by immunofluorescence staining. Protein expression levels of NF-κB p65, p-IκB, NLRP3, NLRC4, ASC, and pro-Caspase-1 in intestinal tissues were analyzed by Western blotting. Cytokine levels of IL-1β, TNF-α, IL-6, and IL-22 in the small intestine after 8 days of administration were determined by Luminex multiplex assay while IL-6 levels were measured by ELISA. Results After 3 days of oral Kai-Xin-San administration, the small intestinal transit rate was significantly reduced in a dose-dependent manner. After 8 days of administration, pathological injuries were observed in the jejunum and ileum, including villus atrophy, mucosal architectural disruption, and luminal dilatation. MUC2 fluorescence kept decreasing with the increase in Kai-Xin-San doses. At 1.5 g·kg^-1, Kai-Xin-San markedly upregulated protein expressions of pro-Caspase-1, NF-κB p65, and p-IκB in the small intestine, but no obvious changes were detected in the colon. After 3 days of treatment, these markers did not change significantly in either the small intestine or colon. Luminex and ELISA results both showed a significant increase in IL-6 levels in the 1.5 g·kg^-1dose group. Conclusion Three-day intragastric administration of Kai-Xin-San at 0.5 g·kg^-1 can induce bloating and intestinal dysmotility in mice. Kai-Xin-San may activate the NF-κB pathway, promote IL-6 transcription, and trigger downstream inflammatory responses. The small intestine, particularly the jejunum and ileum, is the principal target organ, suggesting site-specific intestinal toxicity.

Key words: Kai-Xin-San, Intestinal Inflammation, NF-κB, IL-6, Small Intestinal Motility, Mucus Barrier, Mice

中图分类号:

刘思宇, 杨依霏, 张海静, 张国壮, 宫平, 杨云, 刘婷, 夏冰. 基于NF-κB-IL-6通路探究开心散致小鼠肠道炎症的分子机制[J]. 中国药物警戒, 2026, 23(5): 519-526.

LIU Siyu, YANG Yifei, ZHANG Haijing, ZHANG Guozhuang, GONG Ping, YANG Yun, LIU Ting, XIA Bing. Intestinal Inflammation Induced by Kai-Xin-San through NF-κB-IL-6 Pathway[J]. Chinese Journal of Pharmacovigilance, 2026, 23(5): 519-526.

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