Effect of <italic style="font-style: italic">Changji'an Formula</italic> （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome

WANG Yongfu; KE Wei; XIE Xiangyu; TANG Hongmei; SI Liuze; CHAI Yuna

doi:10.13288/j.11-2166/r.2026.04.016

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Effect of Changji'an Formula （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome

更新时间：2026-02-10

- Effect of Changji'an Formula （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome
- Journal of Traditional Chinese Medicine Vol. 67, Issue 4, Pages: 439-446(2026)
- 作者机构：
  
  1.郑州大学第一附属医院，河南省郑州市建设东路1号，450052
  2.佛山市中医院
  3.广州中医药大学第一临床医学院
  4.广州中医药大学第一附属医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2026.04.016
  CLC：
- Received：04 September 2025，
  
  Revised：2025-11-14，
  
  Published：17 February 2026
- 稿件说明：
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王永福,柯威,谢翔雨等.肠激安方对肝郁脾虚型腹泻型肠易激综合征模型大鼠结肠组织miR-29b-3p/TRAF3/NF-κB/MLCK轴的影响[J].中医杂志,2026,67(04):439-446.

WANG Yongfu,KE Wei,XIE Xiangyu,et al.Effect of Changji'an Formula （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome[J].Journal of Traditional Chinese Medicine,2026,67(04):439-446.
王永福,柯威,谢翔雨等.肠激安方对肝郁脾虚型腹泻型肠易激综合征模型大鼠结肠组织miR-29b-3p/TRAF3/NF-κB/MLCK轴的影响[J].中医杂志,2026,67(04):439-446. DOI： 10.13288/j.11-2166/r.2026.04.016.

WANG Yongfu,KE Wei,XIE Xiangyu,et al.Effect of Changji'an Formula （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome[J].Journal of Traditional Chinese Medicine,2026,67(04):439-446. DOI： 10.13288/j.11-2166/r.2026.04.016.

摘要

目的

从微小RNA-29b-3p（miR-29b-3p）/肿瘤坏死因子受体相关因子3（TRAF3）/核因子κB（NF-κB）/肌球蛋白轻链激酶（MLCK）轴探究肠激安方对肝郁脾虚型腹泻型肠易激综合征（IBS-D）肠道通透性影响的可能机制。

方法

取出生1天的雄性SD窝鼠24只，采用三因素法（母婴分离+醋酸刺激+束缚应激）共6周制备肝郁脾虚型IBS-D大鼠模型。造模成功后随机分为模型组、匹维溴铵组和肠激安方低、高剂量组，每组6只，另取6只同龄未造模SD大鼠作为对照组。肠激安方低、高剂量组分别给予肠激安方溶液16.74、33.48 g/（kg·d）灌胃，匹维溴铵组大鼠给予匹维溴铵片0.018 g/（kg·d）灌胃，对照组同时予10 ml/（kg·d）蒸馏水灌胃。每日1次，连续

14天。灌胃结束后，检测各组大鼠粪便含水量，采用ELISA法检测大鼠血清肠道通透性指标D-乳酸（D-LA）、二胺氧化酶（DAO）和脂多糖（LPS）含量，实时荧光定量PCR法检测大鼠结肠组织

miR

29b

3p、TRAF3

、肿瘤坏死因子α（

TNF

α）、p65、p50、MLCK

mRNA表达，Western Blot法检测结肠组织TRAF3、TNF-α、p65、磷酸化p65（p-p65）、MLCK、肌球蛋白轻链（MLC）、磷酸化肌球蛋白轻链（p-MLC）及紧密连接蛋白［连接黏附因子A（JAM-A）、闭锁蛋白（Occludin）、紧密连接蛋白1（Claudin-1）］蛋白水平。

结果

与对照组比较，模型组大鼠粪便含水量和血清D-LA、DAO、LPS含量均升高，结肠组织JAM-A、Occludin、Claudin-1蛋白水平降低（

＜0.05或

＜0.01），结肠组织

miR

29b

3p、TNF

α、p65、p50、MLCK

mRNA表达升高，

TRAF3

mRNA及蛋白表达降低，TNF-α、MLCK蛋白水平及p-p65/p65、p-MLC/MLC比值升高（

＜0.01）。与模型组比较，各给药组大鼠粪便含水量及血清D-LA、DAO、LPS含量降低，结肠组织

miR

29b

3p、TNF

α、p65、p50、MLCK

mRNA表达降低，

TRAF3

mRNA表达升高；匹维溴铵组、肠激安方高剂量组大鼠结肠组织Occludin、Claudin-1、TRAF3蛋白水平升高，TNF-α、MLCK蛋白水平及p-p65/p65、p-MLC/MLC比值降低（

＜0.05或

＜0.01）。与肠激安方低剂量组比较，肠激安方高剂量组粪便含水量和血清DAO、LPS含量降低（

＜0.01）。与匹维溴铵组比较，肠激安方高剂量组大鼠血清DAO含量降低（

＜0.01）。

结论

肠激安方可能通过调控miR-29b-3p/TRAF3/NF-κB/MLCK轴，从而降低肝郁脾虚型IBS-D模型大鼠肠道通透性，恢复肠道屏障功能。

Abstract

Objective

To explore the potential mechanism of

Changji'an Formula

（肠激安方） on intestinal permeability for rats with diarrhea-predominant irritable bowel syndrome （IBS-D） of liver depression and spleen deficiency syndrome by the microRNA-29b-3p （miR-29b-3p）/tumor necrosis factor receptor-associated factor 3 （TRAF3）/nuclear factor-κB （NF-κB）/myosin light chain kinase （MLCK） axis.

Methods

Twenty-four 1-day-old male Sprague-Dawley （SD） suckling rats were selected， and the IBS-D rat model of liver depression and spleen deficiency syndrome was established via a three-factor method，i.e. maternal separation plus a

cetic acid stimulation and restraint stress， for 6 consecutive weeks. After successful modeling， the rats were randomly divided into a model group， pinaverium bromide group， low-dose and high-dose

Changji'an Formula

groups， with 6 rats in each group. Another 6 age-matched non-modeled SD rats were included as the control group. The low-dose and high-dose

Changji'an Formula

groups were given intragastric administration of

Changji'an Formula

solution at doses of 16.74 g/（kg·d） and 33.48 g/（kg·d）， respectively； the pinaverium bromide group received intragastric administration of pinaverium bromide tablets at 0.018 g/（kg·d）； and the control group was given distilled water at 10 ml/（kg·d） via intragastric gavage. The intervention was conducted once daily for 14 consecutive days. After the gavage treatment， the fecal water content of rats in each group was measured. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of intestinal permeability indicators， including D-lactic acid （D-LA）， diamine oxidase （DAO）， and lipopolysaccharide （LPS）. Quantitative real-time polymerase chain reaction （qRT-PCR） was conducted to determine the mRNA expression levels of

miR

29b

，

TRAF3

， tumor necrosis factor-α （

TNF

），

p65

，

p50

， and

MLCK

in colonic tissues. Western Blot analysis was employed to detect the protein expression levels of TRAF3， TNF-

， p65， phosphorylated p65 （p-p65）， MLCK， myosin light chain （MLC）， phosphorylated MLC （p-MLC）， and tight junction proteins including junctional adhesion molecule-A （JAM-A）， Occludin， and Claudin-1 in colonic tissues.

Results

Compared with the control group， the model group exhibited significantly increased fecal water content and serum levels of D-LA， DAO， and LPS， along with decreased protein expression levels of JAM-A，

Occludin， and Claudin-1 in colonic tissues （

＜0.05 or

＜0.01）. Additionally， in the model group， the mRNA expression levels of

miR

29b

，

TNF

，

p65

，

p50

， and

MLCK

in colonic tissues were up-regulated， while the mRNA and protein expression levels of

TRAF3

were down-regulated； the protein levels of TNF-α and MLCK， as well as the ratios of p-p65/p65 and p-MLC/MLC， significantly elevated （

＜0.01）. Compared with the model group， all treatment groups showed reduced fecal water content and serum levels of D-LA， DAO， and LPS， along with down-regulated mRNA expression levels of

miR

29b

，

TNF

，

p65

，

p50

， and

MLCK

， and up-regulated

TRAF3

mRNA expression in colonic tissues. Moreover， the pinaverium bromide group and high-dose

Changji'an Formula

group presented increased protein levels of Occludin， Claudin-1， and TRAF3， as well as decreased protein levels of TNF-α and MLCK， and reduced ratios of p-p65/p65 and p-MLC/MLC in colonic tissues （

＜0.05 or

＜0.01）. Compared with the low-dose

Changji'an Formula

group， the high-dose group had lower fecal water content and serum levels of DAO and LPS （

＜0.01）. In comparison with the pinaverium bromide group， the high-dose

Changji'an Formula

group showed a significant decrease in serum DAO level （

＜0.01）.

Conclusions

Changji'an Formula

can reduce intestinal permeability and restore intestinal barrier function in IBS-D rats of liver depression and spleen deficiency syndrome by regulating the miR-29b-3p/TRAF3/NF-κB/MLCK axis.

关键词

Keywords

references

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Effect of Changji'an Formula （肠激安方） on the miR-29b-3p/TRAF3/NF-κB/MLCK Axis in Colonic Tissues in Diarrhea-predominant Irritable Bowel Syndrome Model Rat with Liver Depression and Spleen Deficiency Syndrome

DOI：10.13288/j.11-2166/r.2026.04.016

摘要

Abstract

关键词

Keywords

references