Lactobacillus plantarum 299v Prevents Caspase-Dependent Apoptosis In Vitro

Abstract

Selective microbes used as probiotics can enhance epithelial cell protection. We have previously shown that a Lactobacillus plantarum strain 299v (Lp299v) has the ability to induce mucin genes. In the current study, we utilized a cytokine model of inflammation in cell culture to study the modulation of apoptosis by this probiotic. HT-29 cells were pre-incubated with the Lp299v or L. plantarum strain adh- (Lpadh-), a non-adherent derivative of Lp299v. Cells were challenged with a mixture of cytokines (TNF-α, IFN-γ, and IL-1a) to imitate conditions of inflammation. To assess for cell death, we evaluated TUNEL, multi-caspase, and caspase-3 and caspase-7 activity assays. There was a marked decrease in apoptosis as measured by TUNEL(+) cells in samples pre-treated with Lp299v (18.7 ± 4.1%, p < 0.01) and Lpadh- (16.6 ± 3.2%, p < 0.05) prior to cytokine exposure when compared to cells (43.6 ± 6.2%) exposed to the cytokine mixture. Lp299v pre-incubation with HT-29 cells reduced caspase(+) cells in the multi-caspase activity assay (3.6 ± 0.6%, p < 0.05) compared to cells exposed to cytokines (68.9 ± 5.1%) whereas Lpadh- did not (46.8 ± 17.5%, p > 0.05). Similarly, caspase-3, caspase-7 activity was also reduced by Lp299v. Selected probiotics may confer an exogenous protective effect at the mucosal-luminal interface for intestinal epithelial cells via alteration of caspase-dependent apoptotic pathways.

Functional Properties of Food Origin Lactobacillus in the Gastrointestinal Ecosystem—In Vitro Study

“…Also, effect of probiotic Lb . plantarum 299v on cytokine-induced apoptosis was evaluated [42]. The authors demonstrated that cell pre-incubation with Lb .…”

Probiotics and the Gut Immune System: Indirect Regulation

“…Moreover, Dykstra and collaborators showed that the interaction of this probiotic strain with the cells was responsible to enhance the cellular-driven protection via reduction of the caspase pathway activation [ 23 ]. The importance of mucins was also highlighted by other observations demonstrating a reduced number of Goblet cells in inflammatory lesions of the GIT as well as a decreased functional capacity of the mucins to bind pro-inflammatory molecules and to inhibit bacterial binding in the inflamed colon [ 30 , 46 , 47 ].…”

Probiotics in Disease Prevention and Treatment

“…64 L. plantarum (strain 299v) has the capacity to enhance the production and secretion of mucins (MUC2 and MUC3) from human intestinal (HT-29) epithelial cells. 65 Probiotic mixtures also increase MUC2 gene expression and mucin protein secretion in rat colon. 66 L. rhamnosus GG-derived soluble proteins (p40 and p70) ameliorate intestinal injury and inflammation by inhibiting epithelial cell apoptosis 67 and increasing mucin production through transactivation of the epidermal growth factor receptor.…”

“…L. plantarum (strain 299v) has the capacity to enhance the production and secretion of mucins (MUC2 and MUC3) from human intestinal (HT‐29) epithelial cells . Probiotic mixtures also increase MUC2 gene expression and mucin protein secretion in rat colon .…”