Department of Pharmacy, School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield HD1 3DH, UK.
Centre for Natural Products Discovery, School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, UK.
Molecules. 2023 Jan 30;28(3):1317. doi: 10.3390/molecules28031317.
Skimmianine is a furoquinoline alkaloid which is found in the 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 PGE 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 PGE, 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.
蛇菰宁是一种呋喃喹啉生物碱，存在于该属以及芸香科的其他植物中。本研究评估了蛇菰宁对 LPS 激活的 BV-2 小胶质细胞中促炎介质产生的影响。用蛇菰宁（10、20 和 30 μM）处理培养的 BV-2 细胞，然后用 LPS（100ng/mL）刺激。用 ELISA 法测定细胞上清液中 TNFα 和 IL-6 的水平，用 Griess 法和 EIA 分别测定 NO 和 PGE 的水平。用 Western blot 法测定 iNOS、COX-2、磷酸化 p65 和磷酸化 IκBα 的蛋白表达。结果表明，蛇菰宁降低 LPS 诱导的 TNFα、IL-6、NO 和 PGE 的分泌增加，以及 iNOS 和 COX-2 蛋白表达增加。阐明蛇菰宁抗炎活性的机制的实验表明，蛇菰宁显著抑制 LPS 诱导的 NF-κB 介导的荧光素酶活性增加。蛇菰宁预处理还降低了 LPS 诱导的 NF-κB/p65 和 IκBα 蛋白磷酸化增加。此外，蛇菰宁干扰了 NF-κB 与共识序列的结合能力。蛇菰宁预处理可保护 HT-22 细胞免受小胶质细胞条件培养基诱导的毒性，并增加 MAP-2 表达。本研究结果表明，蛇菰宁通过靶向 NF-κB 激活途径抑制 LPS 激活的小胶质细胞中的神经炎症。蛇菰宁还对小胶质细胞条件培养基诱导的神经毒性产生神经保护作用。
