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Neuroprotection by Skimmianine in Lipopolysaccharide-Activated BV-2 Microglia
Folashade A Ogunrinade1, Victoria U Iwuanyanwu1, Satyajit D Sarker2
1Department of Pharmacy, School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield HD1 3DH, UK.
Abstract:
Skimmianine is a furoquinoline alkaloid which is found in the Zanthoxylum genus and also in other plants of the Rutaceae family. This study evaluated the effects of skimmianine on the production of pro-inflammatory mediators in LPS-activated BV-2 microglia. Cultured BV-2 cells were treated with skimmianine (10, 20 and 30 μM), followed by stimulation with LPS (100 ng/mL). Levels of TNFα and IL-6 in cell supernatants were measured using ELISA, while NO and PGE2 levels were evaluated with Griess assay and EIA, respectively. Western blotting was used to determine the protein expression of iNOS, COX-2, phospho-p65 and phospho-IκBα. Results showed that Skimmianine reduced LPS-induced elevated the secretion of TNFα, IL-6, NO, and PGE2, as well as the increased protein expression of iNOS and COX-2. Experiments to elucidate the mechanisms of the anti-neuroinflammatory activity of skimmianine revealed the significant inhibition of LPS-induced increased NF-κB-mediated luciferase activity. Pre-treatment with skimmianine also reduced LPS-induced the increased phosphorylation of NF-κB/p65 and IκBα proteins. Furthermore, skimmianine interfered with the binding capacity of NF-κB to consensus sites. Skimmianine pre-treatment protected HT-22 cells from toxicity induced by microglia-conditioned media, as well as increasing MAP-2 expression. The results of this study suggest that skimmianine inhibits neuroinflammation in LPS-activated microglia by targeting the NF-κB activation pathway. Skimmianine also produced neuroprotection against neurotoxicity induced by microglia-conditioned media.
Insights
Skimmianine, a plant alkaloid, reduces neuroinflammation by inhibiting the NF-κB pathway in activated microglia. This compound also offers neuroprotection against microglia-induced toxicity.
Area of Science:
- Neuroscience
- Pharmacology
- Immunology
Background:
- Neuroinflammation plays a critical role in neurodegenerative diseases.
- Microglia are key immune cells in the central nervous system, and their activation contributes to neuroinflammation.
- Furoquinoline alkaloids, like skimmianine, are found in plants and possess various biological activities.
Purpose of the Study:
- To investigate the anti-neuroinflammatory effects of skimmianine on lipopolysaccharide (LPS)-activated BV-2 microglia.
- To elucidate the underlying molecular mechanisms of skimmianine's action, particularly its impact on the NF-κB pathway.
- To evaluate the neuroprotective potential of skimmianine against microglia-induced neurotoxicity.
Main Methods:
- BV-2 microglia were treated with skimmianine and stimulated with LPS.
- Pro-inflammatory mediators (TNFα, IL-6, NO, PGE2) were quantified using ELISA, Griess assay, and EIA.
- Protein expression of iNOS, COX-2, phospho-p65, and phospho-IκBα was analyzed by Western blotting.
- NF-κB pathway activation was assessed through luciferase activity assays and analysis of protein phosphorylation and DNA binding.
- Neuroprotection was evaluated using HT-22 cells exposed to microglia-conditioned media.
Main Results:
- Skimmianine significantly reduced LPS-induced secretion of TNFα, IL-6, NO, and PGE2 in microglia.
- Skimmianine suppressed the elevated protein expression of iNOS and COX-2 induced by LPS.
- The compound inhibited LPS-induced NF-κB activation, including luciferase activity, p65 and IκBα phosphorylation, and NF-κB DNA binding.
- Skimmianine pre-treatment protected HT-22 cells from neurotoxicity and increased MAP-2 expression.
Conclusions:
- Skimmianine exhibits potent anti-neuroinflammatory effects in LPS-activated microglia by inhibiting the NF-κB signaling pathway.
- Skimmianine demonstrates neuroprotective properties against microglia-mediated neurotoxicity.
- These findings suggest that skimmianine holds therapeutic potential for neuroinflammatory conditions.
