Callerya atropurpurea 通过 TLR4/MyD88 依赖性级联反应抑制体外炎症反应，并改善体内胃损伤和脓毒性休克。

Callerya atropurpurea suppresses inflammation in vitro and ameliorates gastric injury as well as septic shock in vivo via TLR4/MyD88-dependent cascade.

机构信息

Department of Integrative Biotechnology and Biomedical Institute for Convergence at SKKU (BICS), Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon 16419, Republic of Korea.

Department of Biocosmetics, Sungkyunkwan University, Suwon 16419, Republic of Korea.

出版信息

Phytomedicine. 2022 Oct;105:154338. doi: 10.1016/j.phymed.2022.154338. Epub 2022 Jul 19.

DOI:10.1016/j.phymed.2022.154338

PMID:35921773

Abstract

BACKGROUND

Callerya atropurpurea is a traditional plant in a tropical zone discovered to have anti-inflammatory functions.

PURPOSE

we want to investigate the mechanism related to anti-inflammation of C. atropurpurea ethanol extract (Ca-EE) both in vitro and in vivo.

STUDY DESIGN

Murine macrophage cells and mouse models for gastritis and septic shock were conducted to evaluate the abilities of Ca-EE in anti-inflammation.

METHODS

Ca-EE was tested by HPLC and LC-MS/MS. NO outcome was checked by Griess reagent test. Cell viabilities were evaluated using MTT assay. Inflammatory cytokines were determined via RT-PCR and ELISA. The mechanism of Ca-EE in anti-inflammation was investigated by luciferase reporter gene assay and immunoblot in transcription level and protein level respectively. Gastric injury and septic shock administrated with Ca-EE were studied by H&E, PCR, and immunoblot.

RESULTS

Ca-EE significantly decreased LPS-induced NO production, but hardly stimulated the expression of NO itself. It not only showed no cytotoxicity, but also protected cells from LPS damage. Moreover, Ca-EE decreased TLR4 expression, altered MyD88 recruitment and TRAF6, and suppressed the phospho-Src/PI3K/AKT. Ca-EE inhibited downstream signaling P38, JNK and NF-κB. Finally, Ca-EE alleviated HCl/EtOH-induced gastritis and LPS/poly (I:C)-induced septic shock through the previously mentioned signaling cascades.

CONCLUSION

Ca-EE exhibited an integrated and promising mechanism against TLR4-related inflammation, which shows potential for treating gastritis, septic shock, and other inflammatory diseases.

摘要

背景

Callerya atropurpurea 是一种在热带地区发现的传统植物，具有抗炎功能。

目的

我们旨在研究 Callerya atropurpurea 乙醇提取物（Ca-EE）在体外和体内的抗炎相关机制。

研究设计

使用小鼠巨噬细胞和胃炎及脓毒症休克模型来评估 Ca-EE 的抗炎能力。

方法

通过 HPLC 和 LC-MS/MS 对 Ca-EE 进行测试。通过 Griess 试剂试验检测 NO 含量。使用 MTT 测定法评估细胞活力。通过 RT-PCR 和 ELISA 测定炎性细胞因子。通过荧光素酶报告基因测定和免疫印迹分别在转录水平和蛋白质水平上研究 Ca-EE 抗炎的机制。通过 H&E、PCR 和免疫印迹研究 Ca-EE 对胃损伤和脓毒症休克的作用。

结果

Ca-EE 显著降低 LPS 诱导的 NO 产生，但对 NO 本身的表达几乎没有刺激作用。它不仅没有细胞毒性，还能保护细胞免受 LPS 损伤。此外，Ca-EE 降低了 TLR4 的表达，改变了 MyD88 的募集和 TRAF6，并抑制了磷酸化Src/PI3K/AKT。Ca-EE 抑制了下游信号 P38、JNK 和 NF-κB。最后，Ca-EE 通过上述信号通路缓解了 HCl/EtOH 诱导的胃炎和 LPS/poly（I:C）诱导的脓毒症休克。