Surface-layer protein produced by Lactobacillus crispatus JCM 2009 ameliorates lipopolysaccharide-induced inflammation through autophagy cross-talk with the NF-κB signaling pathway

In recent years, studies on immunomodulation by surface-layer proteins (Slps) have mainly focused on Lactobacillus acidophilus, there is little information on Slp from L. crispatus and its intestinal immunomodulatory mechanisms in macrophages. In our study, the anti-inflammatory actions of Slp derived from L. crispatus JCM 2009 and its related molecular mechanisms were investigated. We initially found that incubation with Slp (5–10 μg/mL) for 4 h significantly inhibited nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS-stimulated RAW264.7 cells (P < 0.001). We then found that Slp inhibited the inflammatory response by regulating the PI3K/AKT/mTOR signaling pathway and activating autophagy in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Furthermore, ELISA and Western blotting results demonstrated that the NF-κB signaling pathway positively regulated autophagic activity to inhibit the productions of PGE2 and NO during this inflammatory response. And p65 was identified as a potentially important NF-κB signaling pathway molecule mediating the effects of Slp on the LPS-induced inflammatory response in RAW264.7 cells. Our findings provide the novel perspective that Slp exerts its anti-inflammatory effects through the activation of autophagy, making it a promising bioactive ingredient for the development of functional foods.