Abstract
Purpose: To investigate the anti-neuroinflammatory properties of Carum carvi Linn. (CCE, Umbelliferae) aqueous extract in stimulated BV-2 microglial cells and explore its underlying mechanisms.Methods: Cell viability assessment was performed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MTT) assay. Lipopolysaccharide (LPS) was used to activate BV-2 microglia. Nitric oxide (NO) levels were measured using Griess assay. Inducible NO synthase (iNOS) and cyclooxygenase (COX) levels were evaluated by Western blot analysis. Interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) production were evaluated by enzyme-linked immunosorbent assay (ELISA).Results: CCE alone did not exhibit any signs of cytotoxicity to BV-2 cells up to 200 μg/ml concentration. The LPS-activated excessive release of NO in BV-2 cells was significantly inhibited by CCE (p < 0.001 at 100 μg/mL). CCE also inhibited the production of inflammatory mediators such as iNOS, COX-2, IL-6 and TNF-α (p < 0.05, p < 0.01 and p < 0.001 at 25, 50 and 100 μg/mL, respectively). Further mechanistic study revealed that CCE acts by regulation of nuclear factor kappa-B (NF-κB) signaling pathway in LPS-stimulated BV-2 microglial cells.Conclusion: The results reveal that CCE exhibited its anti-neuroinflammatory effects via regulation of NF-κB signaling. This can be developed as a potential therapeutic target in ameliorating microgliamediated neuroinflammation.Keywords: Carum carvi, Anti-oxidant, Neuroinflammation, Microglia, Nitric oxide, Interleukin
Highlights
Carum carvi Linn from the family Umbelliferae is a globally distributed spice with a long history as a traditional medicinal plant [1]
Earlier reports from our group demonstrated that the aqueous extract of C. carvi fruit exhibited strong anti-oxidant, adaptogenic and memory enhancing effects in in vitro and in vivo experimental models [5, 6]
It is well documented that microglia can be activated in vitro by lipopolysaccharide (LPS) which enhances the production of immunerelated cytotoxic factors and pro-inflammatory cytokines [10] which resembles the in vivo conditions in the central nervous system (CNS)
Summary
Carum carvi Linn from the family Umbelliferae is a globally distributed spice with a long history as a traditional medicinal plant [1]. It is well documented that microglia can be activated in vitro by lipopolysaccharide (LPS) which enhances the production of immunerelated cytotoxic factors and pro-inflammatory cytokines [10] which resembles the in vivo conditions in the CNS. Aqueous extract of C. carvi was evaluated for its protective effects on LPS-activated neuroinflammatory processes in BV-2 microglial cells and explored its mechanisms involved. The C. carvi extract (CCE) was dissolved in sterile distilled water, Cell culture and viability assay. BV-2 cells (2 x 105 cells/mL) were seeded in 6-well plates in 500 μL complete culture medium and treated with the CCE extract at indicated concentrations for 1 h prior to stimulation with. BV-2 microglia cells (1 x 105 cells/well) were cultured on 96 well plates and treated with the CCE extract at indicated concentrations (25, 50 and 100 μg/mL) for 1 h and stimulated with LPS (1 μg/mL).
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