Xianglian Zhixie Tablet Antagonizes Dextran Sulfate Sodium-Induced Ulcerative Colitis by Attenuating Systemic Inflammation and Modulating Gut Microbiota

Abstract

Purpose

Xianglian Zhixie Tablet (XLZXT), a classical traditional Chinese medicine formulation, is commonly used to treat Ulcerative Colitis (UC) in China. However, the therapeutic mechanisms of XLZXT for UC have yet to be fully understood. This study aimed to investigate the curative benefits of XLZXT and its associated mechanisms for healing UC in mice.

Methods

In the present study, the 1% dextran sulfate sodium (DSS) solution was used to establish the UC model in C57BL/6N mice. To investigate the therapeutic effects of XLZXT on DSS-induced UC mice, several parameters were measured, including DAI score, colon length, spleen index, pathological changes in colon tissue, and levels of inflammatory factors in plasma and colon tissue. By investigating the gut microbiota, assessing the levels of intestinal mucosal protein expression, and looking at the proteins involved in the TLR4/MyD88/NF-B p65 signaling pathway, the mechanisms of XLZXT impact on UC were investigated. Mouse feces were examined for patterns of gut microbiota expression using high-throughput sequencing of 16S rRNA.

Results

XLZXT effectively alleviated UC symptoms and colon pathological damage in DSS-induced UC mice. It improved body weight loss, stool consistency, and hematochezia, while also repairing colon damage. Moreover, it down-regulated pro-inflammatory cytokines (such as TNF-α, IL-1β, and IL-6), and up-regulated anti-inflammatory cytokines (such as IL-10). XLZXT also increased the expression of MUC-2, Occludin and ZO-1, while decreasing the expression of NF-κB, MyD88 and TLR4. Additionally, it regulated gut microbiota disorder by increasing the abundance of beneficial bacteria and reducing the adhesion of intestinal harmful bacteria.

Conclusion

XLZXT demonstrated therapeutic effects on DSS-induced UC mice. The mechanisms may be associated with repairing the intestinal mucosal barrier, regulating the TLR4/MyD88/NF-κB p65 signaling pathway, and restoring the balance of gut microbiota.

Plain Language Summary

This is the first study to probe the therapeutic potential of XLZXT for UC. Our findings indicated that XLZXT could effectively alleviate the clinical symptoms of UC and in-depth apparently modify the levels of relevant cytokines in the organism to maintain the system homeostasis. Our results suggested that XLZXT might treat DSS-induced UC by repairing the intestinal mucosa barrier, regulating the TLR4/MyD88/NF-κB p65 signaling pathway, and restoring gut microbiota balance.

Graphical Abstract

Keywords:

• ulcerative colitis
• TCM
• TLR4/MyD88/NF-кB p65 signaling pathway
• gut microbiota

Abbreviations

Akt, Protein kinase B; MUC-2, Mucoprotein-2; BSA, Bovine serum albumin; MyD88, Myeloiddifferentiationfactor88; CD, Crohn’s disease; IL-10, Interleukin-10; CD-14, Endotoxin receptors on Kupffer cell membrane-14; IL-12, Interleukin-12; CMDI, Colon mucosa damage index; JAK2, Janus kinase 2; CRP, C-reactive protein; LPS, Lipopolysaccharide; DAI, Disease activity index; IP-10, Interferon-inducible protein-10; DSS, Dextran sulfate sodium; NC, Nitrocellulose filter membrane; ECL, Enhanced chemiluminescence; NLRP3, Nucleotide-binding oligomerization domain-like receptor protein 3; Elisa, Enzyme-linked immunosorbent assay; NF-κB, Nuclear factor-kappa B; HE, Hematoxylin-eosin; OD, Optical density; HRP, Horseradish peroxidase; OUT, Operational taxonomic units; IBD, Inflammatory bowel disease; PAS, Periodic acid-schiff; IFN-γ, Interferon gamma; PBS, Phosphate buffered saline; IHC, Immunohistochemistry; PCoA, Principal co-ordinates analysis; IL-1β, Interleukin-1β; PCR, Polymerase chain reaction; IL-6, Interleukin-6; PMSF, Pheny methane sulfonyl fluoride; MD-2, Myeloid differentiation protein-2; PRRs, Pathogen-associated molecular pattern; MPO, Myeloperoxidase.Abbreviations: Akt, Protein kinase B; MUC-2, Mucoprotein-2; BSA, Bovine serum albumin; MyD88, Myeloiddifferentiationfactor88; CD, Crohn’s disease; IL-10, Interleukin-10; CD-14, Endotoxin receptors on Kupffer cell membrane-14; IL-12, Interleukin-12; CMDI, Colon mucosa damage index; JAK2, Janus kinase 2; CRP, C-reactive protein; LPS, Lipopolysaccharide; DAI, Disease activity index; IP-10, Interferon-inducible protein-10; DSS, Dextran sulfate sodium; NC, Nitrocellulose filter membrane; ECL, Enhanced chemiluminescence; NLRP3, Nucleotide-binding oligomerization domain-like receptor protein 3; Elisa, Enzyme-linked immunosorbent assay; NF-κB, Nuclear factor-kappa B; HE, Hematoxylin-eosin; OD, Optical density; HRP, Horseradish peroxidase; OUT, Operational taxonomic units; IBD, Inflammatory bowel disease; PAS, Periodic acid-schiff; IFN-γ, Interferon gamma; PBS, Phosphate buffered saline; IHC, Immunohistochemistry; PCoA, Principal co-ordinates analysis; IL-1β, Interleukin-1β; PCR, Polymerase chain reaction; IL-6, Interleukin-6; PMSF, Pheny methane sulfonyl fluoride; MD-2, Myeloid differentiation protein-2; PRRs, Pathogen-associated molecular pattern; MPO, Myeloperoxidase.

Ethics Statement

All experimental procedures were conducted in accordance with “Guidelines for the Management and Use of Laboratory Animals” (Ministry of Science and Technology, China, 2006), and the study was approved by Experimental Animal Ethics Committee of the Research Institute of Beijing Tongrentang Co. LTD with the ethical approval number YJY-2021-030904.

Acknowledgments

This work is supported by the National Key R&D Program of China. We deeply appreciate all the members of our team.

Author Contributions

All authors contributed to data analysis, drafting or revising the article, have agreed on the journal to which the article will be submitted, gave final approval of the version to be published, and agree to be accountable for all aspects of the work.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This work was supported by National key research and development program (2019YFC1711400).