Anti-neuroinflammatory effects of grossamide from hemp seed via suppression of TLR-4-mediated NF-κB signaling pathways in lipopolysaccharide-stimulated BV2 microglia cells
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.
KeywordsHemp seed Lignanamide Grossamide Anti-neuroinflammatory BV2 microglia cells NF-κB
This work was supported by the National Natural Science Foundation of China (Grant No. 81473323); Key R&D program in Shandong Province (No. 2015GSF119025) and the China-Australia Centre for Health Science Research (Grant No. 2015GJ04).
Compliance with ethical standards
Conflict of interest
The authors declare no competing financial interest.
- 13.Cao J, Li Z, Chen J, Li H (2005) Influence of semen cannabis iol on NO, SOD, GSH-Px, MDA in D-galactose-induced Aging Mice Serum. J Sichuan Tradit Chin Med 23(3):29–30Google Scholar
- 14.Wu SH, Guo YY, Zhang ST, Yang Y, Wang R, Zhang XY, Li HB (2015) The anti-aging effect of semen cannabis oil on the sex difference of Bombyx mori L. Pharmacol Clin Chin Mater Med 31:100–103Google Scholar
- 15.Zhang M, Shen Y, Zhu Z, Wang H (1999) Reseach on anti-inflammation, anti-analgesic and anti-thrombus function of semen cannabis. Res Pract Chin Med 13:13–15Google Scholar
- 16.Li G, Cao Y, Wu S, Zhang Z, Zhang Y, Yang Y, Li H (2015) Influence of semen cannabis iol on liqid levels, inflammmatory cytokines and anti-oxidant of aging model mice. Pharmacol Clin Chin Mater Med 31:109–111Google Scholar
- 17.Su J, He H, Shi M, Xiong X, Chen L (2011) Protective effect of semen cannabis iol on learning and memory impairment mice induced by D-galactose. Chin J Clin Pharmacol Ther 16:1332–1339Google Scholar
- 18.Luo J, Zheng T, Mo Z, Wei Q (2003) Effects of extracts of fructus cannabis on learning and memory capacity and related substances of senile mice induced by D-galactose. J Beijing Normal Univ (Nat Sci) 39(3):386–389Google Scholar
- 30.Kim BW, Koppula S, Kumar H, Park JY, Kim IW, More SV, Kim IS, Han SD, Kim SK, Yoon SH, Choi DK (2015) alpha-Asarone attenuates microglia-mediated neuroinflammation by inhibiting NF kappa B activation and mitigates MPTP-induced behavioral deficits in a mouse model of Parkinson’s disease. Neuropharmacology 97:46–57CrossRefPubMedGoogle Scholar
- 33.Cai P, Fu X, Deng AG, Zhan XJ, Cai GM, Li SX (2010) Anti-aging effect of hemp seed oil, protein and lignanamide of bama on old mice. Cent S Pharm 3:003Google Scholar
- 34.Wang X, Yang X, Tang C (2007) Nutritional assessment of Hemp (Cannabis sativa L.) proteins. Mod Food Sci Technol 7:002Google Scholar
- 35.Brenneisen R (2007) Chemistry and analysis of phytocannabinoids and other Cannabis constituents. In: Marijuana and the Cannabinoids, Springer, New York, pp 17–49Google Scholar
- 44.Sung HC, Liang CJ, Lee CW, Yen FL, Hsiao CY, Wang SH, Jiang-Shieh YF, Tsai JS, Chen YL (2015) The protective effect of eupafolin against TNF-alpha-induced lung inflammation via the reduction of intercellular cell adhesion molecule-1 expression. J Ethnopharmacol 170:136–147CrossRefPubMedGoogle Scholar
- 45.Cho KH, Kim DC, Yoon CS, Ko WM, Lee SJ, Sohn JH, Jang JH, Ahn JS, Kim YC, Oh H (2016) Anti-neuroinflammatory effects of citreohybridonol involving TLR4-MyD88-mediated inhibition of NF-κB and MAPK signaling pathways in lipopolysaccharide-stimulated BV2 cells. Neurochem Int 95:55–62CrossRefPubMedGoogle Scholar