Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation

Li-mei GU; Hui LI; Jun-quan XIA; Cheng-yu PAN; Chao GU; Yao-zhou TIAN

doi:10.1007/s11655-021-3343-4

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Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation

Original Article | Updated：2022-02-22

- Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation
- Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation
- Chinese Journal of Integrative Medicine 2022年28卷第2期页码：124-129
- Affiliations：
  
  1.Gastroenterology Department, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing (210028), China
  2.Gastroenterology Department, Jiangsu Province Hospital on Integration of Chinese and Western Medicine, Nanjing (210028), China
  3.Jiangsu Province Academy of Traditional Chinese Medicine,Nanjing (210028), China
- Author bio：
  
  Prof. TIAN Yao-zhou, E-mail:tianyaozhou1960@163.com
- Funds：
  
  the Scientific Research Project of Jiangsu Provincial Administration of Traditional Chinese Medicine(JD2019SZXYB05);Natural Science Foundation of Nanjing University of Chinese Medicine(XZR2020030);National Administration of Traditional Chinese Medicine: 2019 Project of Building Evidence-Based Practice Capacity for Traditional Chinese Medicine(2019XZZX-XH007)
- DOI：10.1007/s11655-021-3343-4
  中图分类号：
- 纸质出版日期：2022-02-01，
  
  网络出版日期：2021-12-07，
  
  录用日期：2021-04-23
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Li-mei GU, Hui LI, Jun-quan XIA, 等. Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation[J]. Chinese Journal of Integrative Medicine, 2022,28(2):124-129.

Li-mei GU, Hui LI, Jun-quan XIA, et al. Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation[J]. Chinese Journal of Integrative Medicine, 2022,28(2):124-129.
Li-mei GU, Hui LI, Jun-quan XIA, 等. Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation[J]. Chinese Journal of Integrative Medicine, 2022,28(2):124-129. DOI： 10.1007/s11655-021-3343-4.

Li-mei GU, Hui LI, Jun-quan XIA, et al. Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation[J]. Chinese Journal of Integrative Medicine, 2022,28(2):124-129. DOI： 10.1007/s11655-021-3343-4.

摘要

目的:

探讨中医经典方剂黄芩汤 (HQD) 对硫酸葡聚糖钠 (DSS) 诱导的溃疡性结肠炎 (UC) 小鼠模型以及肿瘤坏死因子-α (TNF-α) 诱导的人正常肠上皮细胞损伤的保护作用.

方法:

动物实验: 将 30只C57BL/6J 小鼠随机分为5组 (每组

=6)

分别为空白组、模型对照组 (DSS 组) 、黄芩汤低、高剂量组 (HQD-L 组、HQD-H 组) 和阳性药组 (5-ASA组) . 通过自由饮用配置3% 的 DSS 溶液建立小鼠 UC 模型

同时 HQD-L 组、HQD-H组和5-ASA组分别给予100、200mg/kg HQD以及100mg/kg 5-ASA混悬液灌胃

空白组和 DSS 组分别给予等容积的蒸馏水灌胃

给药9天后

测定各组小鼠 DAI 评分、结肠长度、结肠组织病理学评分

并通过 ELISA 法测定血清 IL-6、IL-1β、TNF-α 含量. 细胞实验: 将FHC细胞暴露于HQD (0.6mg/ml) 12小时

然后用TNF-α (10ng/ml) 处理24小时

通过Western blot检测Claudin-4和Occludin等紧密连接 (TJ) 蛋白表达水平

以及p65和 (NF-κB) -α抑制蛋白 (IκBα) 的磷酸化水平.

结果:

动物实验表明: 与空白组相比

DSS 模型组小鼠体重明显下降

DAI 评分、肠道组织病理学评分明显增高; 与DSS组相比

第8天

HQD-H组小鼠的体重下降及DAI评分均有显著改善(

0.05). 此外

HE染色表明

HQD能够显著改善小鼠结肠的病理结构改变; 其中HQD-H组及5-ASA组的肠道组织病理学评分明显下降

与模型组比较具有统计学差异 (

0.05) . 同时HQD-H组和5-ASA组的血清 IL-6、IL-1β、TNF-α 含量明显降低

有统计学意义 (

0.05) . 细胞实验显示

与TNF-α组相比

HQD可上调Occludin和Claudin-4蛋白的表达

并抑制FHC细胞中p65和IκBα磷酸化水平 (

0.05) .

结论:

黄芩汤通过抑制促炎细胞因子的表达明显缓解了DSS诱导的UC小鼠的症状

并通过抑制TNF-α诱导的NF-κB激活维持了FHC细胞中TJ蛋白的稳态

是其发挥抗溃疡性结肠炎的重要作用机制之一.

Abstract

Objective:

To investigate the protective effect of Chinese herbal formula Huangqin Decoction (HQD) on ulcerative colitis mouse model induced by dextran sulphate sodium (DSS) and human intestinal epithelial cell injury induced by tumour necrosis factor-α (TNF-α).

Methods:

In vivo

30 male C57BL/6 mice were divided into 5 groups using a random number table (

=6 per group)

including control

DSS

5-aminosalicylic acid (5-ASA)

HQD low- (HQD-L) and high-dose (HQD-H) groups. The colitis mouse model was established by 3% (w/v) DSS water for 5 days. Meanwhile

mice in the HQD-L

HQD-H and 5-ASA groups were administrated with 100

200 mg/kg HQD or 100 mg/kg 5-ASA

respectively

once daily by gavage. After 9 days of administration

the body weight

disease activity index (DAI) score and colon length of mice were measured

the pathological changes of colons were analyzed by hematoxylin-eosin staining (HE) staining

and the levels of serum interleukin (IL)-6

IL-1β and TNF-α were measured by enzyme linked immunosorbent assay.

In vitro

the human colon epithelial normal cells (FHC cells) were exposed to HQD (0.6 mg/mL) for 12 h and then treated with TNF-α (10 ng/mL) for 24 h. The tight junction (TJ) protein expression levels of Claudin-4 and Occludin

and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB (NF-κB)-α (IκBα) were measured by Western blot.

Results:

In vivo

compared with the DSS group

HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day (

0.05). Moreover

HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice (

0.05). HE staining showed HQD attenuated the pathological changes of colitis mice

and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group (

0.05). Meanwhile

HQD-H and 5-ASA significantly decreased the serum IL-1β

IL-6 and TNF-α levels of mice (

0.05).

In vitro

experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBα levels in FHC cells compared with the TNF-α group (

0.05).

Conclusion:

HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.

关键词

Keywords

Huangqin Decoctionulcerative colitistumour necrosis factor-αnuclear factor kappaB pathwaytight conjunction proteinChinese medicine

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