Anti-neuroinflammatory effects of grossamide from hemp seed via suppression of TLR-4-mediated NF-κB signaling pathways in lipopolysaccharide-stimulated BV2 microglia cells

Abstract

Grossamide, a representative lignanamide in hemp seed, has been reported to possess potential anti-inflammatory effects. However, the potential anti-neuroinflammatory effects and underlying mechanisms of action of grossamide are still unclear. Therefore, the present study investigated the possible effects and underlying mechanisms of grossamide against lipopolysaccharide (LPS)-induced inflammatory response in BV2 microglia cells. BV2 microglia cells were pre-treated with various concentrations of grossamide before being stimulated with LPS to induce inflammation. The levels of pro-inflammatory cytokines were determined using the enzyme-linked immunoassay (ELISA) and mRNA expression levels were measured by real-time PCR. The translocation of nuclear factor-kappa B (NF-κB) and contribution of TLR4-mediated NF-κB activation on inflammatory effects were evaluated by immunostaining and Western blot analysis. This study demonstrated that grossamide significantly inhibited the secretion of pro-inflammatory mediators such as interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α), and decreased the level of LPS-mediated IL-6 and TNF-α mRNA. In addition, it significantly reduced the phosphorylation levels of NF-κB subunit p65 in a concentration-dependent manner and suppressed translocation of NF-κB p65 into the nucleus. Furthermore, grossamide markedly attenuated the LPS-induced expression of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88). Taken together, these data suggest that grossamide could be a potential therapeutic candidate for inhibiting neuroinflammation in neurodegenerative diseases.

Hempseed Lignanamides Rich-Fraction: Chemical Investigation and Cytotoxicity towards U-87 Glioblastoma Cells

“…Based on previous MS observations, and UV-DAD spectra, compounds 27, and 29, whose pseudomolecular ions were in accordance with the C 36 H 36 N 2 O 8 molecular formula, and showing the base peak at m/z 460.1799(1803), and its demethylated radical ion at m/z 445.1569(1), were suggested to be aryldihydronaphtalene-type lignanamide isomers, whereas compound 37 was tentatively identified as cannabisin F, and compound 39 was putatively assigned as grossamide ( Figure S14). This latter compound, whose MS/MS fragmentation pattern in positive ion mode was previously reported [43], was found to exert anti-neuroinflammatory action, being able to inhibit the secretion of pro-inflammatory mediators (e.g., IL-6, and TNF-α), reducing LPS-mediated IL-6 and TNF-α mRNA levels [44]. Furthermore, neuroprotection by cannabisin F was ascertained against LPS-induced inflammatory response and oxidative stress in BV2 microglia cells [21].…”

Anti-Inflammatory and Anti-Migratory Activities of Isoquinoline-1-Carboxamide Derivatives in LPS-Treated BV2 Microglial Cells via Inhibition of MAPKs/NF-κB Pathway

“…It is known that LPS can stimulate and activate TLR-4, a member of the TLR family, expressed on the surface of microglia. Upon the activation of TLR-4 by LPS, an intracellular adapter protein, MyD88, is recruited and, subsequently, both the NF-κB and MAPKs pathways are activated, producing various pro-inflammatory mediators [57]. Thus, inhibition of TLR-4-mediated signaling molecule(s) may be beneficial to counteract inflammation.…”

Therapeutic effect and mechanism of action of quercetin in a rat model of osteoarthritis

“…4 Toll-like receptor (TLR)-4/nuclear factor kappa-light-chainenhancer of activated B cells (NF-jB) signaling is a critical pathway in the development of inflammation. [5][6][7] However, the efficacy and mechanism of action of quercetin in OA remain unclear. In the present study, we developed a rat model of OA by intra-articular injection of papain.…”