Remimazolam attenuates intestinal barrier dysfunction and metabolic disorder in mice with traumatic brain injury by regulating macrophage M2 polarization and restoring tight junction proteins

Cai Xinrong; Li Weichao; Zhang Chen; She Han; Mao Qingxiang

doi:10.16016/j.2097-0927.202607047

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Remimazolam attenuates intestinal barrier dysfunction and metabolic disorder in mice with traumatic brain injury by regulating macrophage M2 polarization and restoring tight junction proteins

Monographic Reports | 更新时间：2026-09-16

- Remimazolam attenuates intestinal barrier dysfunction and metabolic disorder in mice with traumatic brain injury by regulating macrophage M2 polarization and restoring tight junction proteins
- Journal of Army Medical University Vol. 48, Issue 17, Pages: 2428-2439(2026)
- 作者机构：
  
  陆军军医大学（第三军医大学）大坪医院麻醉科，重庆
- 作者简介：
  
  Mao Qingxiang，E-mail：qxmao@tmmu.edu.cn
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(82671675);the Key Project of Natural Science Foundation of Chongqing(2024NSCQ-KJFZZDX0047);the Project for Innovative Leading Talents of Chongqing Talent Program(CSTC2024Y-CJH-BGZXM0011);the Chongqing medical scientific research project （Joint project of Chongqing Health Commission and Science and Technology Bureau）(2025MSXM042)
- DOI：10.16016/j.2097-0927.202607047
  CLC： R333.3;R651.15;R971.2
- Received：13 July 2026，
  
  Revised：2026-08-30，
  
  Accepted：31 August 2026，
  
  Published：15 September 2026
- 稿件说明：
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蔡欣蓉，李维超，张琛，等.瑞马唑仑调控巨噬细胞M2极化修复紧密连接蛋白而减轻创伤性颅脑损伤小鼠肠道屏障功能障碍及代谢紊乱［J］.陆军军医大学学报，2026，48（17）：2428-2439.DOI：10.16016/j.2097-0927.202607047.

Cai X R，Li W C，Zhang C，et al.Remimazolam attenuates intestinal barrier dysfunction and metabolic disorder in mice with traumatic brain injury by regulating macrophage M2 polarization and restoring tight junction proteins［J］.J Army Med Univ，2026，48（17）：2428-2439.DOI：10.16016/j.2097-0927.202607047.
蔡欣蓉，李维超，张琛，等.瑞马唑仑调控巨噬细胞M2极化修复紧密连接蛋白而减轻创伤性颅脑损伤小鼠肠道屏障功能障碍及代谢紊乱［J］.陆军军医大学学报，2026，48（17）：2428-2439.DOI：10.16016/j.2097-0927.202607047. DOI：

Cai X R，Li W C，Zhang C，et al.Remimazolam attenuates intestinal barrier dysfunction and metabolic disorder in mice with traumatic brain injury by regulating macrophage M2 polarization and restoring tight junction proteins［J］.J Army Med Univ，2026，48（17）：2428-2439.DOI：10.16016/j.2097-0927.202607047. DOI：

摘要

目的

创伤性颅脑损伤（traumatic brain injury，TBI）可导致肠道屏障损伤及代谢紊乱。瑞马唑仑（remimazolam，Remi）具有一定的抗炎和器官保护作用。本研究拟观察Remi对TBI小鼠肠道屏障及肠道代谢的影响，并探索其可能的保护机制。

方法

120只8~12周龄雄性C57BL/6小鼠（体质量18~20 g）采用随机数字法分为Sham组（仅行开颅术）、TBI组（采用控制性皮质撞击法造模）和TBI+Remi组（于造模前2 h腹腔注射瑞马唑仑10 mg/kg），每组40只，造模后24 h取材。HE染色检测回肠组织病理形态学改变，采用Chiu评分标准对回肠组织损伤程度进行半定量评价，透射电子显微镜观察回肠上皮细胞超微结构，伊文思蓝（evans blue，EB）染色检测肠道通透性，Western blotting检测紧密连接蛋白［闭锁蛋白（Occludin）、闭锁小带蛋白-1（zonula occludens-1，ZO-1）］的表达水平，ELISA检测回肠组织屏障功能及黏膜免疫相关指标［二胺氧化酶（diamine oxidase，DAO）、D-乳酸脱氢酶（D-lactate dehydrogenase，D-LDH）、分泌型免疫球蛋白A（secretory immunoglobulin A，sIgA）］的水平；Western blotting和免疫荧光检测诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）和甘露糖受体（cluster of differentiation 206，CD206）表达。采用非靶向代谢组学分析肠道代谢变化，并进行差异代谢物筛选及KEGG通路富集分析。

结果

与Sham组相比，TBI组回肠黏膜及超微结构明显受损，Chiu评分、肠道通透性及DAO、D-LDH、sIgA水平升高，Occludin和ZO-1表达降低（

0.05）；与TBI组相比，TBI+Remi组上述改变均明显改善（

0.05）。与Sham组相比，TBI组iNOS表达升高、CD206表达降低；与TBI组相比，TBI+Remi组iNOS表达下降、CD206表达升高（

0.05）。非靶向代谢组学共鉴定2 567种代谢物，与Sham组相比，TBI组筛选出851种差异代谢物，其中260种上调、591种下调，主要涉及氨基酸代谢、不饱和脂肪酸及胆汁酸代谢、糖代谢和核苷酸代谢等通路；TBI组瓜氨酸和酪氨酸相对丰度降低，5-氨基乙酰丙酸（5-aminolevulinic acid，5-ALA）

和内源性前列腺素E2代谢物（15-Keto-prostaglandin E2，15-Keto-PGE2）升高；与TBI组相比，TBI+Remi组瓜氨酸和酪氨酸相对丰度升高，而5-ALA和15-Keto-PGE2相对丰度降低（

0.05）。

结论

瑞马唑仑可减轻TBI小鼠肠黏膜及上皮超微结构损伤，增强紧密连接蛋白表达并降低肠道通透性，同时部分改善TBI所致的肠道代谢紊乱。

Abstract

Objective

Traumatic brain injury (TBI) can lead to intestinal barrier disruption and metabolic disorder. Remimazolam (Remi) possesses certain anti-inflammatory and organ-protective effects. This study aims to determine the effects of Remi on intestinal barrier function and intestinal metabolism in TBI mice and explore its potential protective mechanisms.

Methods

A total of 120 male C57BL/6 mice (8 to 12 weeks old

weighing 18 to 20 g) were randomly divided into the Sham group (craniotomy only)

the TBI group (established using controlled cortical impact)

and the TBI+Remi group (intraperitoneal administration of 10 mg/kg Remi 2 h before modelling)

with 40 mice in each group. Intestinal tissue samples were collected 24 h after modeling. Histopathological changes in ileal tissues were observed by HE staining

and the degree of ileal injury was semi-quantitatively assessed using the Chiu’s scoring system. Transmission electron microscopy was used to observe the ultrastructure of ileal epithelial cells

and Evans blue (EB) staining was employed to assess intestinal permeability. Western blotting was performed to detect the expression levels of tight junction proteins [Occludin and zonula occludens-1 (ZO-1)]. ELISA was applied to measure the levels of intestinal barrier function-related and mucosal immune-related indicators

including diamine oxidase (DAO)

D-lactate dehydrogenase (D-LDH)

and secretory immunoglobulin A (sIgA). The expression levels of inducible nitric oxide synthase (iNOS) and cluster of differentiation 206 (CD206) were detected by Western blotting and immunofluorescence assay. Untargeted metabolomics was conducted to analyze intestinal metabolic changes

followed by differential metabolite screening and KEGG pathway enrichment analysis.

Results

Compared with the Sham group

the TBI group showed significantly damaged ileal mucosa and ultrastructure

with increased Chiu’s score

intestinal permeability

and levels of DAO

D-LDH

and sIgA

as well as decreased expression of Occludin and ZO-1 (

0.05). Remi treatment resulted in significant improvements in all the above changes compared with the condition in the TBI group (

0.05). Increased iNOS expression and decreased CD206 expression were observed in the TBI group compared to the Sham group

while Remi treatment reversed the changes induced by TBI modeling (

0.05). Untargeted metabolomics identified 2 567 metabolites

and compared with the Sham group

851 differential metabolites were found in the TBI group

including 260 upregulated and 591 downregulated metabolites

mainly involved in amino acid metabolism

unsaturated fatty acid and bile acid metabolism

glucose metabolism

and nucleotide metabolism pathways. The relative abundance of citrulline and tyrosine was decreased in the TBI group

whereas 5-aminolevulinic acid (5-ALA) and the endogenous prostaglandin E2 metabolite 15-keto-prostaglandin E2 (15-keto-PGE2) were increased. Compared with the TBI group

the TBI+Remi group showed increased relative abundance of citrulline and tyrosine and decreased relative abundance of 5-ALA and 15-keto-PGE2 (

0.05).

Conclusion

Remi alleviates intestinal mucosal and epithelial ultrastructural damage

enhances tight junction protein expression

and reduces intestinal permeability in TBI mice

and partially ameliorates TBI-induced intestinal metabolic disturbances.

关键词

Keywords

references

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