Soluble Fraction from Lysate of a High Concentration Multi-Strain Probiotic Formulation Inhibits TGF-β1-Induced Intestinal Fibrosis on CCD-18Co Cells

Abstract

Fibrosis is a severe complication of chronic inflammatory disorders, such as inflammatory bowel disease (IBD). Current strategies are not fully effective in treating fibrosis; therefore, innovative anti-fibrotic approaches are urgently needed. TGF-β1 plays a central role in the fibrotic process by inducing myofibroblast differentiation and excessive extracellular matrix (ECM) protein deposition. Here, we explored the potential anti-fibrotic impact of two high concentration multi-strain probiotic formulations on TGF-β1-activated human intestinal colonic myofibroblast CCD-18Co. Human colonic fibroblast CCD-18Co cells were cultured in the presence of TGF-β1 to develop a fibrotic phenotype. Cell viability and growth were measured using the Trypan Blue dye exclusion test. The collagen-I, α-SMA, and pSmad2/3 expression levels were evaluated by Western blot analysis. Fibrosis markers were also analyzed by immunofluorescence and microscopy. The levels of TGF-β1 in the culture medium were assessed by ELISA. The effects of commercially available probiotic products VSL#3® and Vivomixx® were evaluated as the soluble fraction of bacterial lysates. The results suggested that the soluble fraction of Vivomixx® formulation, but not VSL#3®, was able to antagonize the pro-fibrotic effects of TGF-β1 on CCD-18Co cells, being able to prevent all of the cellular and molecular parameters that are related to the fibrotic phenotype. The mechanism underlying the observed effect appeared to be associated with inhibition of the TGF-β1/Smad signaling pathway. To our knowledge, this study provides the first experimental evidence that Vivomixx® could be considered to be a promising candidate against intestinal fibrosis, being able to antagonize TGF-β1 pro-fibrotic effects. The differences that were observed in our fibrosis model between the two probiotics used could be attributable to the different number of strains in different proportions.

Bifidobacterium bifidum FJSWX19M5 alleviated 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced chronic colitis by mitigating gut barrier injury and increasing regulatory T cells

“…31 A recent report showed that the lysates of multi-strain probiotics inhibited TGF-β1-induced intestinal fibrosis. 32…”

“…31 A recent report showed that the lysates of multi-strain probiotics inhibited TGF-β1-induced intestinal fibrosis. 32 The protective effects of the specific probiotics can be partly attributed to their regulatory effects on the intestinal barrier, including TJ function and mucin production. Herein, the specific B. bifidum strain (FJSWX19M5) increased TJ production and improved the physical barrier in the colons of TNBS-treated mice (Fig.…”

Therapeutic Targeting of Intestinal Fibrosis in Crohn’s Disease

“…Given the increasing emphasis on the pro-fibrotic impact of the gut microbiota, many in vitro and in vivo studies have been carried out to assess the effect of probiotics and prebiotics on intestinal fibrosis [134][135][136][137]. Among these, the most recent suggested that the soluble fraction of Vivomixx ® formulation was able to inhibit collagen-I and α-SMA expression in human colonic fibroblast by interfering TGF-β1/Smad2-3 signaling [138]. All the data available are still preliminary and need to be confirmed and expanded.…”

The Probiotic Strains Bifidοbacterium lactis, Lactobacillus acidophilus, Lactiplantibacillus plantarum and Saccharomyces boulardii Regulate Wound Healing and Chemokine Responses in Human Intestinal Subepithelial Myofibroblasts

“…In addition, these cells both express a wide range of cytokine receptors and produce immunological mediators and growth factors, for instance TGF-β, and are therefore involved not only in wound re-epithelization but also in the inflammatory and fibrotic cascades of the gut [ 47 , 50 , 51 ]. In DSS-induced colitis in mice, treatment with Lactiplantibacillus plantarum has shown a myofibroblast-mediated anti-inflammatory and anti-fibrotic effect [ 52 ], while in another in vitro study utilizing human myofibroblasts, a mix of eight different probiotic bacteria, including Lactobacillus and Bifidobacterium strains, has proven efficacy in TGF-β inhibition [ 53 ]. Apart from offering their innate properties, i.e., the regulation of PGE 2 production by cultured myofibroblasts stimulated with Lactobacillus rhamnosus [ 54 ] and intestinal organoids stimulated with Lactobacillus acidophilus [ 55 ], recombinant probiotics could even be used as regulatory protein domain carriers [ 56 ].…”