花椒麻素通过抑制 NF-κB 转录因子和 NLRP3 炎性小体的激活抑制脂多糖和人工血影蛋白诱导的 BV-2 小胶质细胞神经炎症。

Zanthoxylum zanthoxyloides inhibits lipopolysaccharide- and synthetic hemozoin-induced neuroinflammation in BV-2 microglia: roles of NF-κB transcription factor and NLRP3 inflammasome activation.

机构信息

Department of Pharmacy, School of Applied Sciences, University of Huddersfield, Huddersfield, UK.

Centre for Natural Products Discovery, School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, Byrom Street, Liverpool, UK.

出版信息

J Pharm Pharmacol. 2021 Mar 1;73(1):118-134. doi: 10.1093/jpp/rgaa019.

DOI:10.1093/jpp/rgaa019

PMID:33791805

Abstract

OBJECTIVES

The effects of a root extract of Zanthoxylum zanthoxyloides on neuroinflammation in BV-2 microglia stimulated with LPS and hemozoin were investigated.

METHODS

ELISA, enzyme immunoassay and Griess assay were used to evaluate levels of cytokines, PGE2 and NO in culture supernatants, respectively. Microglia-mediated neurotoxicity was evaluated using a BV-2 microglia-HT-22 neuron transwell co-culture.

KEY FINDINGS

Treatment with Z. zanthoxyloides caused reduced elevated levels of TNFα, IL-6, IL-1β, NO and PGE2, while increasing the levels of IL-10. In addition, there were reduced levels of iNOS and COX-2 proteins. This was accompanied by a prevention of microglia-mediated damage to HT-22 mouse hippocampal neurons. Z. zanthoxyloides reduced elevated levels of phospho-IκB and phospho-p65, while preventing degradation of IκB protein and DNA binding of p65. Further mechanistic studies revealed that Z. zanthoxyloides reduced the levels of pro-IL-1β and IL-1β in hemozoin-activated BV-2 microglia. This was accompanied by a reduction in caspase-1 activity and NLRP3 protein expression. Bioassay-guided fractionation resulted in the isolation of skimmianine as an anti-inflammatory compound in Z. zanthoxyloides.

CONCLUSION

This is the first report showing the inhibition of neuroinflammation in LPS- and hemozoin-activated BV-2 microglia by the root extract of Z. zanthoxyloides by targeting the activation of both NF-κB and NLRP3 inflammasome.

摘要

目的

研究两面针根提取物对 LPS 和血晶素刺激的 BV-2 小胶质细胞神经炎症的影响。

方法

采用 ELISA、酶免疫测定和格里斯测定法分别评估细胞培养上清液中细胞因子、PGE2 和 NO 的水平。通过 BV-2 小胶质细胞-HT22 神经元共培养评估小胶质细胞介导的神经毒性。

主要发现

两面针根提取物处理可降低 TNFα、IL-6、IL-1β、NO 和 PGE2 的升高水平，同时增加 IL-10 的水平。此外，iNOS 和 COX-2 蛋白水平降低。这伴随着对 HT-22 鼠海马神经元的小胶质细胞介导损伤的预防。两面针根提取物降低了磷酸化 IκB 和磷酸化 p65 的升高水平，同时防止 IκB 蛋白降解和 p65 的 DNA 结合。进一步的机制研究表明，两面针根提取物降低了血晶素激活的 BV-2 小胶质细胞中促 IL-1β 和 IL-1β 的水平。这伴随着 caspase-1 活性和 NLRP3 蛋白表达的降低。基于生物测定的分级分离导致从两面针根提取物中分离出 skimmianine 作为一种抗炎化合物。