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通过靶向NF-κB途径研究枸杞(Lycium barbarum L.)叶和嫩枝甲醇提取物的抗炎功能

Anti-Inflammatory Functions of Methanol Extract from (L.) Borkh. Leaves and Shoots by Targeting the NF-κB Pathway.

作者信息

Song Chaoran, Chen Hongxi, Kim Soo Ah, Lee Jong Sub, Choung Eui Su, Zhang Zhiyun, Kim Soo-Yong, Kim Jong-Hoon, Cho Jae Youl

机构信息

Department of Integrative Biotechnology, Biomedical Institute for Convergence at SKKU (BICS), Sungkyunkwan University, Suwon 16419, Korea.

DanjoungBio Co., Ltd., Wonju 26303, Korea.

出版信息

Plants (Basel). 2022 Feb 26;11(5):646. doi: 10.3390/plants11050646.

Abstract

(L.) Borkh. is a widely used medical plant in Asia. Since the anti-inflammatory mechanism of this plant is not fully understood, the aim of this study was to explore the anti-inflammatory function and mechanism of (L.) Borkh. methanol extract (Mb-ME). For in vitro experiments, nitric oxide production assay, PCR, overexpression strategy, immunoblotting, luciferase reporter assay, and immunoprecipitation were employed to explore the molecular mechanism and the target proteins of Mb-ME. For in vivo experiments, an HCl/EtOH-induced gastritis mouse model was used to confirm the anti-inflammatory function. Mb-ME showed a strong ability to inhibit the production of nitric oxide and the expression of inflammatory genes. Mb-ME decreased NF-κB luciferase activity mediated by MyD88 and TRIF. Moreover, Mb-ME blocked the activation of Src, Syk, p85, Akt, p50, p60, IKKα/β, and IκBα in LPS-induced RAW264.7 cells. Overexpression and immunoprecipitation analyses suggested Syk and Src as the target enzymes of Mb-ME. In vitro results showed that Mb-ME could alleviate gastritis and relieve the protein expression of p-Src, p-Syk, and COX-2, as well as the gene expression of COX-2 and TNF-α. In summary, this study implied that Mb-ME performs an anti-inflammatory role by suppressing Syk and Src in the NF-κB signaling pathway, both in vivo and in vitro.

摘要

(L.) Borkh.是一种在亚洲广泛使用的药用植物。由于该植物的抗炎机制尚未完全明确,本研究旨在探究(L.) Borkh.甲醇提取物(Mb-ME)的抗炎功能及机制。在体外实验中,采用一氧化氮生成测定、聚合酶链反应、过表达策略、免疫印迹、荧光素酶报告基因测定和免疫沉淀等方法来探究Mb-ME的分子机制和靶蛋白。在体内实验中,使用盐酸/乙醇诱导的胃炎小鼠模型来证实其抗炎功能。Mb-ME表现出强大的抑制一氧化氮生成和炎症基因表达的能力。Mb-ME降低了由MyD88和TRIF介导的NF-κB荧光素酶活性。此外,Mb-ME阻断了脂多糖诱导的RAW264.7细胞中Src、Syk、p85、Akt、p50、p60、IKKα/β和IκBα的激活。过表达和免疫沉淀分析表明Syk和Src是Mb-ME的靶酶。体外实验结果显示,Mb-ME可减轻胃炎,并降低p-Src、p-Syk和COX-2的蛋白表达以及COX-2和TNF-α的基因表达。总之,本研究表明Mb-ME在体内和体外均通过抑制NF-κB信号通路中的Syk和Src发挥抗炎作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10e7/8912290/5290ec23541b/plants-11-00646-g001a.jpg

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