Lactobacillus paracasei-derived extracellular vesicles attenuate the intestinal inflammatory response by augmenting the endoplasmic reticulum stress pathway

Abstract

Lactobacillus paracasei is a major probiotic and is well known for its anti-inflammatory properties. Thus, we investigated the effects of L. paracasei-derived extracellular vesicles (LpEVs) on LPS-induced inflammation in HT29 human colorectal cancer cells and dextran sulfate sodium (DSS)-induced colitis in C57BL/6 mice. ER stress inhibitors (salubrinal or 4-PBA) or CHOP siRNA were utilized to investigate the relationship between LpEV-induced endoplasmic reticulum (ER) stress and the inhibitory effect of LpEVs against LPS-induced inflammation. DSS (2%) was administered to male C57BL/6 mice to induce inflammatory bowel disease, and disease activity was measured by determining colon length, disease activity index, and survival ratio. In in vitro experiments, LpEVs reduced the expression of the LPS-induced pro-inflammatory cytokines IL-1 alpha, IL-1 beta, IL-2, and TNF alpha and increased the expression of the anti-inflammatory cytokines IL-10 and TGF beta. LpEVs reduced LPS-induced inflammation in HT29 cells and decreased the activation of inflammation-associated proteins, such as COX-2, iNOS and NF kappa B, as well as nitric oxide. In in vivo mouse experiments, the oral administration of LpEVs also protected against DSS-induced colitis by reducing weight loss, maintaining colon length, and decreasing the disease activity index (DAI). In addition, LpEVs induced the expression of endoplasmic reticulum (ER) stress-associated proteins, while the inhibition of these proteins blocked the anti-inflammatory effects of LpEVs in LPS-treated HT29 cells, restoring the pro-inflammatory effects of LPS. This study found that LpEVs attenuate LPS-induced inflammation in the intestine through ER stress activation. Our results suggest that LpEVs have a significant effect in maintaining colorectal homeostasis in inflammation-mediated pathogenesis. Inflammatory bowel disease: Vesicles from gut bacteria quell inflammation Tiny vesicles released by a bacterial species found in the human gut can reduce symptoms of inflammatory bowel disease (IBD) and prevent disease progression. People with IBD have a decreased abundance of Lactobacilli bacteria in their gut, creating an imbalance that perpetuates the disease. Replenishment of this bacteria may become a valuable therapy. Chang Mo Moon at Ewha Womans University, Yoon-Keun Kim at MD Healthcare, both in Seoul, South Korea, and co-workers demonstrated how extracellular vesicles (EVs) released by Lactobacilli paracasei can actively prevent bowel inflammation. These EVs contain a mixture of proteins, nucleic acids and other biomolecules. The team administered EV to cultured human colorectal cancer cells and to mice with induced colitis. The EVs decreased pro-inflammatory protein activity and boosted levels of protective cellular membrane proteins via augmenting ER stress pathway.