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Journal of Traditional Chinese Medicine >

2024 , Vol. 44 >Issue 5: 885 - 895

DOI: https://doi.org/10.19852/j.cnki.jtcm.20240806.006

Regulatory effects of the p38 mitogen-activated protein kinase-myosin light chain kinase pathway on the intestinal epithelial mechanical barrier and the mechanism of modified Pulsatilla decoction (加味白头翁汤) in the treatment of ulcerative colitis

  • Tingting WU ,
  • Xin YANG ,
  • Huiping ZHU ,
  • Jinwei GUO ,
  • Hui ZHU ,
  • Peipei ZHANG ,
  • Meng WANG ,
  • Guoqiang LIANG ,
  • Hongwen SUN
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  • 1 Department of Internal Medicine, Suzhou TCM Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou 215003, China
    2 Department of Internal Medicine, The Affiliated Suzhou Science and Technology Town Hospital of Nanjing Medical University, Suzhou 215153, China
    3 Suzhou Academy of Wumen Chinese Medicine, Suzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, Suzhou 215003, China

Received date: 2023-06-11

  Accepted date: 2023-11-15

  Online published: 2024-08-06

Supported by

Natural Science Foundation of Jiangsu Province-funded Project: Regulation of Intestinal Epithelial Tight Junction Proteins Through the p38 Mitogen-activated Protein Kinase-Myosin Light Chain kinase Signaling Pathway in Ulcerative Colitis Treatment by Modified Pulsatilla Decoction(BK20201179);Jiangsu Health Commission-funded Project: Study on the Prevention of Postoperative Recurrence in Patients with Colorectal Adenoma and High Triglyceride based on the Theory of “Treating the Disease Before It Gets Worse” by Modified Jisheng Wumei Pill(BJ23039);Suzhou Health Commission-funded Project: Study on the Technical Application of Xu Jingfan's “Eliminating Stasis and Protecting Membrane” Method to Regulate Recombinant NLR Family, Pyrin Domain Containing Protein 3 to Repair Esophageal Mucosa in Reflux Esophagitis treatment(SS202080);Suzhou Health Commission-funded Project(SKY2022015);Suzhou Health Commission-funded Project: Clinical Diagnostic and Therapeutic Techniques for Reflux Esophagitis Treatment with Graded Step-down Therapy by Combining Traditional Chinese and Western medicine(LCZX201817)

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Abstract

OBJECTIVE: To investigate the mechanism of the protective effect of modified Pulsatilla decoction (加味白头翁汤, MPD) on the mechanical barrier of the ulcerative colitis (UC) intestinal epithelium in vitro and in vivo.

METHODS: We established an intestinal epithelial crypt cell line-6 cell barrier injury model by using lipopolysaccharide (LPS). The model was then treated with p38 mitogen-activated protein kinase-myosin light chain kinase (p38MAPK-MLCK) pathway inhibitors, p38MAPK-MLCK pathway silencing genes (si-p38MAPK, si-NF-κB, and si-MLCK), and MPD respectively. Transepithelial electronic resistance (TEER) measurements and permeability assays were performed to assess barrier function. Immunofluorescence staining of tight junctions (TJ) was performed. In in vivo experiment, dextran sodium sulfate-induced colitis rat model was conducted to evaluate the effect of MPD and mesalazine on UC. The rats were scored using the disease activity index based on their clinical symptoms. Transmission electron microscopy and hematoxylin-eosin staining were used to examine morphological changes in UC rats. Western blotting and real-time quantitative polymerase chain reaction were performed to examine the gene and protein expression of significant differential molecules.

RESULTS: In in vitro study, LPS-induced intestinal barrier dysfunction was inhibited by p38MAPK-MLCK pathway inhibitors and p38MAPK-MLCK pathway gene silencing. Silencing of p38MAPK-MLCK pathway genes decreased TJ expression. MPD treatment partly restored the LPS-induced decreased in TEER and increase in permeability. MPD increased the gene and protein expression of TJ, while down-regulated the LPS-induced high expression of p-p38MAPK and p-MLC. In UC model rats, MPD could ameliorate body weight loss and disease activity index, relieve colonic pathology, up-regulate TJ expression as well as decrease the expression of p-p38MAPK and p-MLC in UC rat colonic mucosal tissue.

CONCLUSIONS: The p38MAPK-MLCK signaling pathway can affect mechanical barrier function and TJ expression in the intestinal epithelium. MPD restores TJ expression and attenuates intestinal epithelial barrier damage by suppressing the p38MAPK-MLCK pathway.

Key words： colitis, ulcerative; mitogen-activated protein kinase 14; myosin-light-chain kinase; modified pulsatilla decoction; intestinal epithelial mechanical barrier; experimental verification

Cite this article

Tingting WU , Xin YANG , Huiping ZHU , Jinwei GUO , Hui ZHU , Peipei ZHANG , Meng WANG , Guoqiang LIANG , Hongwen SUN . Regulatory effects of the p38 mitogen-activated protein kinase-myosin light chain kinase pathway on the intestinal epithelial mechanical barrier and the mechanism of modified Pulsatilla decoction (加味白头翁汤) in the treatment of ulcerative colitis[J]. Journal of Traditional Chinese Medicine, 2024 , 44(5) : 885 -895 . DOI: 10.19852/j.cnki.jtcm.20240806.006

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