Weissella confusa WIKIM51 ameliorates constipation and improves gut health in loperamide-induced mice

다국어 초록 (Multilingual Abstract)

Constipation is a prevalent gastrointestinal disorder that significantly impacts physical well-being and daily quality of life. In recent years, probiotics have gained increasing attention as promising alternatives to conventional laxatives, which often cause undesirable side effects and dependency. Weissella confusa WIKIM51 (Wilac D), a strain originally isolated from dandelion kimchi, has previously demonstrated anti-obesity properties; however, its potential role in relieving constipation has not yet been elucidated. The aim of this study was to determine whether Wilac D could alleviate constipation symptoms in mice induced with loperamide. Male ICR mice were orally administered either vehicle, bisacodyl (positive control), or low and high doses of Wilac D (1×109 and 1×1010 CFU/mouse, respectively) for four weeks. Constipation was induced by loperamide during the last 11 days of treatment. Wilac D supplementation markedly improved fecal parameters, including stool weight, water content, and defecation frequency, while reducing fecal pH and residual pellets in the colon. In addition, Wilac D restored tight junction gene expression and suppressed the transcription of water channel proteins, including AQP3, AQP4, and AQP8. Furthermore, Wilac D modulated gastrointestinal hormones associated with motility, decreasing Somatostatin (SS) and Vasoactive intestinal peptide (VIP) levels while increasing Gastrin (Gas), Substance P (SP), and Cholecystokinin (CCK). The expression of c-Kit and SCF was assessed by western blot analysis, revealing an increase that indicates activation of interstitial cells of Cajal signaling. 16S rRNA amplicon–based metagenomic analysis revealed restoration of microbial balance and increased abundances of A. muciniphila, B. acidifaciens, Weissella, and S24-7. Collectively, Wilac D mitigates constipation by improving stool properties, regulating intestinal motility related hormones, strengthening the gut barrier, and modulating microbial composition. Wilac D may therefore represent a safe, effective, and functional approach for promoting intestinal health and managing constipation.

목차 (Table of Contents)

• I. Introduction 1
• II. Materials and Methods 8
• 1. Strain preparation 8
• 2. Experiment design 9
• 3. Assessment of fecal defecation-related indicators 10
• 4. Gastrointestinal transit time (GITT) 11
• 5. Small intestinal transit time (SIT) 11
• 6. Measurement of serum biochemistry markers 12
• 7. Histopathological evaluation 13
• 8. RNA extraction and qRT-PCR analysis 14
• 9. Protein extraction and western blot analysis 16
• 10. Microbiota analysis 18
• 11. Statistical analysis 19
• III. Results 20
• 1. Wilac D modulates defecation-related parameters in mice with LP- induced constipation 20
• 2. Wilac D enhances intestinal motility in mice with LP-induced constipation 23
• 3. Wilac D modulates GI peptides and hormones in mice with LP- induced constipation 25
• 4. Wilac D alters intestinal structure and enhances mucus secretion in mice with LP-induced constipation 27
• 5. Wilac D modulates genes involved in intestinal barrier function in mice with LP-induced constipation 31
• 6. Wilac D regulates intestinal motility-related signaling in mice with LP-induced constipation 35
• 7. Wilac D modulates gut microbial communities in mice with LP- induced constipation 38
• 8. Wilac D alters the differential abundance of gut microbial taxa in mice with LP-induced constipation 41
• IV. Discussion 44
• V. Conclusion 51
• Reference 52
• 국문 초록 61