Baicalein mitigates time-dependent colonic barrier injury in mice after high-intensity exercise in hot and humid environment by inhibiting oxidative stress and inflammation: A study on intervention dosage and critical time window

WU Xiaolong; LIU Yurong; LIU Yuqing; LI Linfang; GOU Xinyue; YANG Xuesen; YI Long; MI Mantian; CHEN Ka

doi:10.16016/j.2097-0927.202511059

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Baicalein mitigates time-dependent colonic barrier injury in mice after high-intensity exercise in hot and humid environment by inhibiting oxidative stress and inflammation: A study on intervention dosage and critical time window

Military Medicine | 更新时间：2026-03-30

- Baicalein mitigates time-dependent colonic barrier injury in mice after high-intensity exercise in hot and humid environment by inhibiting oxidative stress and inflammation: A study on intervention dosage and critical time window
- Journal of Army Medical University Vol. 48, Issue 6, Pages: 670-686(2026)
- 作者机构：
  
  1.陆军军医大学（第三军医大学）军事预防医学系：，军队营养与食品卫生学教研室，重庆市营养与健康重点实验室，重庆市医学营养研究中心，重庆
  2.陆军军医大学（第三军医大学）军事预防医学系：，热带医学教研室，重庆
- 作者简介：
  
  MI Mantian, E-mail：mantian_mi@tmmu.edu.cn
  CHEN Ka, E-mail：chenka@tmmu.edu.cn
- 基金信息：
  
  the High-Temperature Training Support （2024）.
- DOI：10.16016/j.2097-0927.202511059
  CLC： R285.5;R574.62;G804.2
- Received：11 November 2025，
  
  Revised：2026-01-27，
  
  Published：30 March 2026
- 稿件说明：
移动端阅览
伍小龙，柳玉容，刘渝清，等.黄芩素通过抑制氧化应激和炎症减轻湿热环境高强度运动后小鼠结肠屏障时序性损伤：干预剂量与关键时间窗研究［J］.陆军军医大学学报，2026，48（06）：670-686.DOI：10.16016/j.2097-0927.202511059.

WU X L，LIU Y R，LIU Y Q，et al.Baicalein Mitigates Time-Dependent Colonic Barrier Injury in Mice After High-Intensity Exercise in Hot and Humid Environment by Inhibiting Oxidative Stress and Inflammation： A Study on Intervention Dosage and Critical Time Window［J］.J Army Med Univ，2026，48（06）：670-686.DOI：10.16016/j.2097-0927.202511059.
伍小龙，柳玉容，刘渝清，等.黄芩素通过抑制氧化应激和炎症减轻湿热环境高强度运动后小鼠结肠屏障时序性损伤：干预剂量与关键时间窗研究［J］.陆军军医大学学报，2026，48（06）：670-686.DOI：10.16016/j.2097-0927.202511059. DOI：

WU X L，LIU Y R，LIU Y Q，et al.Baicalein Mitigates Time-Dependent Colonic Barrier Injury in Mice After High-Intensity Exercise in Hot and Humid Environment by Inhibiting Oxidative Stress and Inflammation： A Study on Intervention Dosage and Critical Time Window［J］.J Army Med Univ，2026，48（06）：670-686.DOI：10.16016/j.2097-0927.202511059. DOI：

摘要

目的

本研究旨在明确单次模拟湿热环境高强度运动负荷后小鼠结肠屏障损伤特征及时序规律，并进一步探讨黄芩素（baicalein，BAI）干预对结肠损伤的防护效应，为制定湿热环境下高强度运动及作业人员的医学防护时间窗与药物干预方案提供实验依据。

方法

①SPF级8周龄雄性C57BL/6J小鼠30只，按随机数字表法分为5组（

=6）：对照组，运动后30 min组、4 h组、12 h组及24 h组，运动组小鼠在热舱内（温度：38 ℃、湿度：RH75%）的跑台上，以80%最大运动速度（Vmax）跑步（运动12 min、休息8 min，循环3次；坡度10°）。随后取小鼠结肠组织用HE染色观察各组形态学变化；ELISA检测血清中炎症因子（TNF-α、IL-6、IL-10和IL-1β）水平观察各组炎症反应，检测肠脂肪酸结合蛋白（I-FABP）、脂多糖（LPS）观察肠道通透性；微量法测定结肠中还原型谷胱甘肽（GSH）、丙二醛（MDA）浓度，过氧化氢酶（CAT）、超氧化物歧化酶（SOD）活性；RT-qPCR检测结肠紧密连接蛋白相关基因

Occludin、ZO-1、Claudin-1

，炎症及氧化应激相关基因

Tlr4、TNF-α、IL-1β、IL-6、IL-10、HO-1

（血红素加氧酶-1）

、SOD1

（超氧化物歧化酶1），以及热应激调控中关键转录因子

HSF1、NF-κB、PARP-1、NRF2

的表达水平；Western blot检测结肠紧密连接蛋白Occludin的含量。②黄芩素干预研究，SPF级8周龄雄性C57BL/6J小鼠24只，按随机数字表法分为4组（

=6）：对照组、运动组（去离子水灌胃200 μL）、运动+低剂量干预组［BAI灌胃200 μL，10 mg/（kg·d）］、运动+高剂量干预组［BAI灌胃200 μL， 40 mg/（kg·d）］，于运动结束后4 h取小鼠结肠及血液样本，检测血清尿素氮（BUN）含量、谷丙转氨酶（ALT）及谷草转氨酶（AST）活性评价黄芩素的药物安全性，其余检

测的部位、指标及方法同前。

结果

①运动组小鼠核心体温显著升高（

0.05）；结肠黏膜出现结构性损伤并伴有炎性细胞浸润；结肠

Occludin、ZO-1、Claudin-1

表达下调（

0.05），Occludin蛋白含量降低（

0.05），血清I-FABP及LPS浓度升高（

0.05）；血清TNF-α、IL-1β、IL-6、IL-10水平升高（

0.05），结肠

IL-1β、IL-10

表达上调（

0.05）；结肠MDA浓度升高（

0.05），GSH浓度、CAT及SOD活性降低（

0.05）；结肠

HSF1

、

PARP-1、NF-κB、Tlr4、TNF-α、IL-6

、

SOD1

表达上调（

0.05），

NRF2、HO-1

表达下调（

0.05）。上述变化在运动后30 min即存在，运动后4 h最为显著，24 h则明显恢复。②黄芩素干预后小鼠核心体温显著下降（

0.05），血清ALT及AST浓度降低（

0.05）；结肠黏膜损伤减轻；结肠

Occludin

基因表达及其蛋白含量上升（

0.05），

ZO-1、Claudin-1

表达上调（

0.05），血清I-FABP、LPS浓度降低（

0.05）；血清IL-1β水平降低（

0.05），结肠

IL-1β

表达下调（

0.05）；结肠MDA浓度降低（

0.05），GSH浓度、CAT及SOD活性升高（

0.05）；结肠

HSF1、PARP-1、NF

κB、Tlr4、TNF-α、IL-6

表达下调（

0.05），

NRF2、HO-1

表达上调（

0.05）。上述结果表明，黄芩素具有抗炎抗氧化作用，且低剂量组的效果更好。

结论

模拟湿热环境高强度运动后4 h小鼠结肠屏障出现峰值损伤，预防性使用黄芩素［10 mg/（kg·d）］可以显著抑制炎症与氧化应激并减轻损伤，为热应激导致的结肠损伤提供时间窗证据及干预依据。

Abstract

Objective

This study aims to clarify the characteristics and temporal patterns of colonic barrier damage in mice following a single simulated high-intensity exercise in hot and humid environment

and further investigate the protective effects of baicalein (BAI) intervention on colonic injury

thereby providing experimental evidence for the development of medical protection time windows and pharmaceutical intervention protocols for personnel engaged in high-intensity exercise and operations in hot and humid environments.

Methods

①Thirty SPF-grade 8-week-old male C57BL/6J mice were randomly divided into 5 groups (

=6): control group

30 min post-exercise group

4 h post-exercise group

12 h post-exercise group

and 24 h post-exercise group. The mice in the exercise groups were subjected on a treadmill in a heat chamber (temperature: 38℃

humidity: RH 75%) at 80% maximal exercise velocity (Vmax) (exercise 12 min

rest 8 min

3 cycles; slope 10°). Subsequently

the colon tissues were harvested for morphological observation by HE staining in each group; ELISA was used to detect the serum levels of inflammatory factors (TNF-α

IL-6

IL-10

and IL-1β) to observe the inflammatory response of each group. Intestinal fatty acid binding protein (I-FABP) and lipopolysaccharide (LPS) were detected to observe intestinal permeability; Microassay was performed to determine the concentrations of reduced glutathione (GSH) and malondialdehyde (MDA)

as well as the activity of catalase (CAT) and superoxide dismutase (SOD) in the colon tissues; RT-qPCR was applied to detect the expression levels of intestinal tight junction protein related genes

Occludin

ZO-1

Claudin-1

inflammation and oxidative stress related genes

Tlr4

TNF-α

IL-1β

IL-6

IL-10

heme oxygenase-1 (

HO-1

)

and superoxide dismutase1 (

SOD1

)

as well as key transcription factors

HSF1

NF-κB

PARP-1

and

NRF2

in heat stress regulation; Western blotting was utilized to detect the protein of colonic tight junction protein Occludin. ② For BAI intervention study

24 SPF-grade 8-week-old male C57BL/6J mice were randomly divided into 4 groups (

=6) control group

exercise group (high temperature and high humidity exercise

gavage with deionized water 200 μL)

exercise+low- and high-dose intervention groups (BAI gavage 200 μL at 10 and 40 mg/kg per day). At 4 h after the end

of exercise

colonic and blood samples were collected for serum levels of urea nitrogen (BUN) content

alanine aminotransferase (ALT)

and aspartate aminotransferase (AST) to evaluate the safety of BAI. Other detection sites

indicators

and methods were the same as aforementioned.

Results

① The core body temperature was significantly elevated in the exercise group (

0.05); the structural damage

along with inflammatory cell infiltration was observed in colonic mucosa; the expression of colon

Occludin

ZO-1

and

Claudin-1

was downregulated (

0.05)

and the protein level of Occludin was decreased (

0.05)

and the serum levels of I-FABP and LPS were increased (

0.05); The serum levels of TNF-α

IL-1β

IL-6

and IL-10 were increased (

0.05)

and the expression of

IL-1β

and

IL-10

in the colon was upregulated (

0.05); The concentration of MDA was increased (

0.05)

while the concentration of GSH and the activities of CAT and SOD were decreased (

0.05); The expression of

HSF1

PARP-1

NF-κB

Tlr4

TNF-α

IL-6

and

SOD1

in the colon was upregulated (

0.05)

while the expression of

NRF2

and

HO-1

was downregulated (

0.05). These changes were present at 30 min post-exercise

most prominent at 4 h post-exercise

and markedly recovered by 24 h. ② After BAI intervention

the core body temperature was significantly decreased (

0.05)

and the serum levels of ALT and AST were decreased (

0.05); colonic mucosal injury was alleviated; the expression of Occludin at mRNA and protein levels in the colonic tissues were increased (

0.05); the expression of

ZO-1

and

Claudin-1

were upregulated (

0.05)

and serum levels of I-FABP and LPS were decreased (

0.05); the serum level of IL-1β was decreased (

0.05)

and the expression of

IL-1β

in the colon was downregulated (

0.05); The concentration of MDA was decreased (

0.05)

while the concentration of GSH and activities of CAT and SOD in the colon were increased (

0.05); The expression of

HSF1

PARP-1

NF-κB

Tlr4

TNF-α

and

IL-6

in the colon was downregulated (

0.05)

while the expression of

NRF2

and

HO-1

was upregulated (

0.05). These results indicated that BAI possesses anti-inflammatory and antioxidant effects

with better efficacy in the low-dose group.

Conclusion

Peak colonic barrier injury occurs at 4 h after simulated high-intensity exercise in a hot and humid environment

and prophylactic use of BAI (10 mg/kg per day) can significantly inhibit inflammation and oxidative stress and alleviate injury

providing evidence for time windows and intervention basis for colonic injury induced by heat stress.

关键词

Keywords

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