Every-other-day fasting alleviates secondary neuroinflammation after spinal cord injury by modulating gut microbiota to promote microglial M2 polarization

HUANG Shiyu; LIU Jiancheng; ZHANG Xin; WANG Wenchun; ZHU Li; YANG Yi; WANG Linjie; PANG Rizhao

doi:10.16016/j.2097-0927.202602035

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Every-other-day fasting alleviates secondary neuroinflammation after spinal cord injury by modulating gut microbiota to promote microglial M2 polarization

Basic Medicine | 更新时间：2026-07-15

- Every-other-day fasting alleviates secondary neuroinflammation after spinal cord injury by modulating gut microbiota to promote microglial M2 polarization
- Journal of Army Medical University Vol. 48, Issue 13, Pages: 1863-1875(2026)
- 作者机构：
  
  1.西南交通大学医学院，四川成都
  2.中国人民解放军西部战区总医院康复医学科，四川成都
  3.四川省中医骨科与运动康复临床医学研究中心，四川成都
- 作者简介：
  
  WANG Linjie， E-mail：1554671237@qq.com
  PANG Rizhao， E-mail：przprz17@126.com
- 基金信息：
  
  the General Project of Natural Science Foundation of Sichuan Province(2025ZNSFSC0774);the Project of Technology Innovation Research and Development of Chengdu(2024-YF05-02486-SN)
- DOI：10.16016/j.2097-0927.202602035
  CLC： R378.2;R459.3;R745.44
- Received：05 February 2026，
  
  Revised：2026-03-06，
  
  Accepted：30 March 2026，
  
  Published：15 July 2026
- 稿件说明：
移动端阅览
黄诗宇，刘建成，张鑫，等.隔日限食通过调控肠道菌群促进小胶质细胞M2型活化减轻脊髓损伤后继发性神经炎症［J］.陆军军医大学学报，2026，48（13）：1863-1875.DOI：10.16016/j.2097-0927.202602035.

HUANG S Y，LIU J C，ZHANG X，et al.Every-other-day fasting alleviates secondary neuroinflammation after spinal cord injury by modulating gut microbiota to promote microglial M2 polarization［J］.J Army Med Univ，2026，48（13）：1863-1875.DOI：10.16016/j.2097-0927.202602035.
黄诗宇，刘建成，张鑫，等.隔日限食通过调控肠道菌群促进小胶质细胞M2型活化减轻脊髓损伤后继发性神经炎症［J］.陆军军医大学学报，2026，48（13）：1863-1875.DOI：10.16016/j.2097-0927.202602035. DOI：

HUANG S Y，LIU J C，ZHANG X，et al.Every-other-day fasting alleviates secondary neuroinflammation after spinal cord injury by modulating gut microbiota to promote microglial M2 polarization［J］.J Army Med Univ，2026，48（13）：1863-1875.DOI：10.16016/j.2097-0927.202602035. DOI：

摘要

目的

脊髓损伤（spinal cord injury，SCI）后肠道菌群紊乱可通过肠-脑轴影响小胶质细胞活化并加剧继发性神经炎症，隔日限食（every-other-day fasting，EODF）虽能重塑肠道菌群、促进小胶质细胞M2型活化并改善SCI后神经炎症，但其是否依赖肠道菌群发挥作用尚不明确。本研究基于伪无菌（gut microbiota depletion，GMD）模型与粪菌移植（fecal microbiota transplantation，FMT）模型，探讨EODF能否通过调控肠道菌群促进SCI后小胶质细胞M2型活化，进而减轻继发性炎症。

方法

采用随机数字表法对89只5~6周龄，体质量160~180 g的健康雄性SPF级SD大鼠进行分组。GMD组（饮用联合抗生素水，

=6）与正常饮食（ad libitum，AL）组（饮用灭菌水，

=6）干预14 d后采集大鼠粪便，通过16S rDNA高通量测序进行GMD模型验证；通过正常饮食（AL组，

=12）与隔日限食（EODF组，

=20）干预4周后分别制备正常饮食粪菌移植液（AL-FMT）和隔日限食粪菌移植液（EODF-FMT）；45只大鼠采用C5右侧半钳夹的方法在GMD模型的基础上建立SCI模型，分为假手术（Sham）组、正常饮食（AL-SCI）组、隔日限食（EODF-SCI）组、移植正常菌群（AL-FMT-SCI）组及移植隔日限食菌群（EODF-FMT-SCI）组，每组9只，干预28 d。通过16S rDNA高通量测序分析肠道菌群；每周记录各组大鼠的摄食量与体质量；采用梳理试验评估上肢运动功能；ELISA法检测血清脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）、IL-4、脂多糖（lipopolysaccharide，LPS）水平；Western blotting检测M1型小胶质细胞标记物iNOS和M2型小胶质细胞Arg-1的相对表达及脊髓组织炎症相关蛋白IL-1β、TNF-α的相对表达。

结果

与AL组相比，GMD组的α与β多样性均显著降低（

0.05），在科水平上，GMD组

Bacteroidaceae

等菌群相对丰度均显著降低（

0.05），GMD大鼠模型构建成功；在SCI模型中，与EODF-SCI组和AL-SCI组相比，EODF-FMT-SCI组的产短链脂肪酸（short-chain fatty acids，SCFAs）相关菌群相对丰度增加，且与AL-FMT-SCI组相比，EODF-FMT-SCI组显著增加了

Bacteroidaceae

、

Lactobacillus gasseri

等有益菌的相对丰度；EODF-FMT-SCI组术后14 d体质量与术后7 d形成显著差异（

0.05）；术后14 d，EODF-FMT-SCI组的梳理实验评分已较术后第3天显著提高（

0.05），术后7、14、28 d，EODF-FMT-SCI组梳理实验评分显著高于AL-SCI组（

0.05）；与EODF-SCI组和AL-SCI组相比，EODF-FMT-SCI组血清IL-4、BDNF含量显著增加，LPS含量降低，脊髓组织中IL-1β蛋白与TNF-α蛋白相对表达下降（

0.05），EODF-FMT-SCI组脊髓组织iNOS蛋白相对表达明显下调，Arg-1蛋白相对表达明显上调（

0.05），且EODF-FMT-SCI组脊髓组织Arg-1蛋白相对表达水平显著高于AL-FMT-SCI组（

0.05）。

结论

EODF可能通过重塑肠道菌群，促进SCI后小胶质细胞M2型活化，从而减轻继发性炎症，促进运动功能恢复。

Abstract

Objective

Gut microbiota dysbiosis after spinal cord injury (SCI) can affect microglial activation and exacerbate secondary neuroinflammation

via

the gut-brain axis. Although every-other-day fasting (EODF) reshapes gut microbiota

promotes microglial polarization toward the M2 phenotype

and ameliorates neuroinflammation after SCI

whether it exerts these effects depending on gut microbiota remains unclear. Using a pseudo-germ-free (gut microbiota depletion

GMD) model and fecal microbiota transplantation (FMT)

this study aims to investigate whether EODF promotes microglial M2 polarization after SCI by modulating gut microbiota

thereby alleviating secondary inflammation.

Methods

A total of 89 healthy male SPF grade SD rats (age 5 to 6 weeks

weighing 160 to 180 g) were randomly divided into groups. The GMD group (drinking antibiotic cocktail

=6) and the ad libitum (AL) group (drinking sterilized water

=6) were treated for 14 d before rat feces were collec

ted for 16S rDNA high-throughput sequencing to validate the GMD model. Normal diet fecal microbiota transplantation (AL-FMT) and EODF-FMT were prepared after 4 weeks of intervention with normal diet (AL

=12) and EODF (

=20)

respectively. 45 rats were subjected to C5 right-sided hemiclamp SCI modeling on the basis of the GMD model

and then divided into a Sham group

an AL-SCI group

an EODF-SCI group

an AL-FMT-SCI group

and an EODF-FMT-SCI group

with 9 rats in each group. All of the above interventions lasted for 28 d. Gut microbiota was analyzed by 16S rDNA high-throughput sequencing. Food intake and body weight of rats in each group were recorded weekly. Upper limb motor function was evaluated using grooming test. Serum levels of brain-derived neurotrophic factor (BDNF)

IL-4

and lipopolysaccharide (LPS) were measured by ELISA. The protein levels of the M1 microglial marker iNOS and the M2 microglial marker Arg-1

as well as the levels of the spinal cord tissue inflammation-related proteins IL-1β and TNF-α

were detected by Western blotting.

Results

Compared with the AL group

both α-diversity and β-diversity were significantly decreased in the GMD group (

0.05). At the family level

the relative abundances of

Bacteroidaceae

and other microbiota were markedly reduced in the GMD group (

0.05)

indicating successful establishment of the GMD model. In the SCI model

compared with the EODF-SCI and AL-SCI groups

the relative abundances of short-chain fatty acids (SCFAs)-producing microbiota were increased in the EODF-FMT-SCI group

whereas compared with the AL-FMT-SCI group

the EODF-FMT-SCI group exhibited significantly increased relative abundances of beneficial bacteria such as

Bacteroidaceae

and

Lactobacillus gasseri

. A significant difference in body weight was observed in the EODF-FMT-SCI group between 14 and 7 d postoperatively (

0.05). I

n the grooming test

the scores at 7 d postoperatively were significantly higher in the EODF-FMT-SCI group than those at 3 d after surgery (

0.05)

and the scores at 7

14 and 28 d after surgery in the EODF-FMT-SCI group were obviously higher than those in the AL-SCI group at corresponding time points (

0.05). Compared with the EODF-SCI and AL-SCI groups

the EODF-FMT-SCI group showed significantly increased serum levels of IL-4 and BDNF

decreased LPS content

and reduced protein levels of IL-1β and TNF-α in the spinal cord tissues (

0.05)

downregulated expression of iNOS and upregulated expression of Arg-1 in the spinal cord tissues (

0.05). Furthermore

the expression level of Arg-1 protein in the spinal cord tissue was significantly higher of the EODF-FMT-SCI group than that of the AL-FMT-SCI group (

0.05).

Conclusion

EODF may alleviate secondary inflammation and promote motor functional recovery by reshaping gut microbiota and promoting microglial M2 polarization after SCI.

关键词

Keywords

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