绒毛钩吻通过抑制Src/Syk 改善胃和肺损伤。

Cocculus hirsutus ameliorates gastric and lung injuries by suppressing Src/Syk.

机构信息

Department of Integrative Biotechnology, Sungkyunkwan University, Suwon 16419, Republic of Korea.

AmorePacific R&D Center, Yongin, 17074, Republic of Korea.

出版信息

Phytomedicine. 2021 Dec;93:153778. doi: 10.1016/j.phymed.2021.153778. Epub 2021 Sep 27.

DOI:10.1016/j.phymed.2021.153778

PMID:34628239

Abstract

BACKGROUND

Cocculus hirsutus (L.) W. Thedo., a traditionally well-known plant, has confirmed antitumor properties as well as acute and chronic diuretic effects. However, little is known about its inflammatory activities and the potential effect on inflammatory disease treatment.

PURPOSE

Our aim in this study was to explore additional beneficial properties of C. hirsutus ethanol extract (Ch-EE) such as anti-inflammatory activity in vitro and in vivo as well as its underlying mechanisms and to provide a theoretical basis for its role as a candidate natural drug in clinical gastritis and lung disease therapy.

STUDY DESIGN

RAW264.7 cells, HEK293T cells, peritoneal macrophages, and mouse models of acute gastritis and acute lung injury were used to assess the anti-inflammatory activity of Ch-EE.

METHODS

Decreases in LPS-induced nitric oxide (NO) production and cytokine expression by RAW264.7 cells after Ch-EE treatment were evaluated by Griess assays and PCR, respectively. Transcription factor activity was assessed through luciferase reporter gene assay, and protein expression was determined by Western blotting analysis. Overexpression assays and cellular thermal shift assays were executed in HEK293T cells. Our two in vivo models were an HCl/EtOH-induced gastritis model and an LPS-induced lung injury model. Changes in stomach lesions, lung edema, and lung histology were examined upon treatment with Ch-EE. Components of Ch-EE were determined by liquid chromatography-mass spectrometry.

RESULTS

LPS-induced nitric oxide production and Pam3CSK4- and L-NAME-induced NO production were inhibited by Ch-EE treatment of RAW264.7 cells. Furthermore, LPS-induced increases in transcript levels of iNOS, COX2, CCL12, and IL-1β were reduced by Ch-EE treatment. Ch-EE decreased both MyD88- and TRIF-induced NF-κB promotor activity. Proteins upstream of NF-κB, namely p-p50, p-p65, p-IκBα, p-AKT1, p-Src, and p-Syk, were all downregulated by Ch-EE. Moreover, Src and Syk were targets of Ch-EE. Ch-EE treatment reduced the size of inflammatory stomach lesions induced by HCl/EtOH, lung edema, and accumulation of activated neutrophils caused by LPS.

CONCLUSIONS

These results strongly suggest that Cocculus hirsutus can be developed as a promising anti-inflammatory remedy with Src- and Syk-inhibitory functions targeting diseases related to gastritis and lung injury.

摘要

背景

毛诃子（L.）W. Thedo.，一种传统上广为人知的植物，已被证实具有抗肿瘤特性以及急性和慢性利尿作用。然而，人们对其炎症活动及其在炎症性疾病治疗中的潜在作用知之甚少。

目的

本研究旨在探索毛诃子乙醇提取物（Ch-EE）的其他有益特性，如体外和体内的抗炎活性及其潜在机制，并为其作为候选天然药物在临床胃炎和肺部疾病治疗中的作用提供理论依据。

研究设计

使用 RAW264.7 细胞、HEK293T 细胞、腹腔巨噬细胞以及急性胃炎和急性肺损伤的小鼠模型来评估 Ch-EE 的抗炎活性。

方法

通过 Griess 测定法和 PCR 分别评估 Ch-EE 处理后 LPS 诱导的 RAW264.7 细胞中一氧化氮（NO）产生和细胞因子表达的降低。通过荧光素酶报告基因测定法评估转录因子活性，并通过 Western blot 分析测定蛋白表达。在 HEK293T 细胞中进行过表达测定和细胞热转移测定。我们的两个体内模型是盐酸/乙醇诱导的胃炎模型和 LPS 诱导的肺损伤模型。通过 Ch-EE 处理观察胃病变、肺水肿和肺组织学的变化。通过液相色谱-质谱法确定 Ch-EE 的成分。

结果

Ch-EE 处理 RAW264.7 细胞可抑制 LPS 诱导的一氧化氮产生和 Pam3CSK4 和 L-NAME 诱导的一氧化氮产生。此外，Ch-EE 处理可降低 LPS 诱导的 iNOS、COX2、CCL12 和 IL-1β 的转录水平。Ch-EE 降低了 MyD88 和 TRIF 诱导的 NF-κB 启动子活性。NF-κB 的上游蛋白，即 p-p50、p-p65、p-IκBα、p-AKT1、p-Src 和 p-Syk，均被 Ch-EE 下调。此外，Src 和 Syk 是 Ch-EE 的靶点。Ch-EE 处理可减轻 HCl/EtOH 诱导的炎症性胃病变、肺水肿以及 LPS 引起的活化中性粒细胞的积聚。