Antimicrobial peptide CC34 attenuates intestinal inflammation via downregulation of the NF-κB signaling pathway.

The investigational drug CC34 is a cation peptide with multiple bioactivities. Here, we studied the anti-inflammatory effects of CC34 in lipopolysaccharide (LPS)-treated mouse monocyte-macrophage cells (RAW264.7) and in mice with LPS-induced intestinal inflammation. In vitro, CC34 treatment with less than 50 μg/mL for 24 h did not induce cytotoxicity in RAW264.7 cells. Furthermore, CC34 significantly lowered the levels of select inflammatory cytokines, including TNF-α, IL-1β, and IL-6. Intracellular levels of reactive oxygen species (ROS) were lower in RAW264.7 cells treated with CC34 + LPS than in cells treated with LPS alone. Additionally, CC34 treatment suppressed iNOS and COX-2 mRNA levels in LPS-treated cells. We also observed that CC34 exerted anti-inflammatory activity by suppressing the phosphorylation of IKKβ, IκBα, and NF-κB p65 in vitro. Moreover, CC34 downregulated the release of inflammatory cytokines (TNF-α, IL-1β, and IL-6) in the jejunum tissue and serum of LPS-treated mice. We also found that the myeloperoxidase (MPO) levels were decreased, and the pathological damages were effectively abated in the jejunum tissue of CC34 + LPS-treated mice. In summary, we demonstrated that CC34 exerted anti-inflammatory activities, associated with the neutralization of LPS, inhibition of ROS, inhibition the NF-κB signaling pathway, and down-regulating the secretion of inflammatory cytokines. Thus, CC34 may represent an effective therapeutic strategy for intestinal inflammation. © King Abdulaziz City for Science and Technology 2021.