Wen Quan, Lau Ngaikeung, Weng Huandi, Ye Peng, Du Shaohui, Li Chun, Lv Jianping, Li Hui
Guangdong-HongKong-Macau Institute of CNS Regeneration, Jinan University, Guangzhou, China.
School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Front Bioeng Biotechnol. 2021 Jan 25;8:623866. doi: 10.3389/fbioe.2020.623866. eCollection 2020.
Chrysophanol (Chr) is the main monomer isolated from . This study aimed to identify the potential and cytoprotective effects of Chr on lipopolysaccharide (LPS)-triggered acute lung injury (ALI). We used an ALI-murine model and constructed an inflammatory macrophage cell model to determine the cellular mechanisms involved in Chr-mediated activity. To observe the vital role of histone deacetylase 3 (HDAC3) in abolishing inflammation action, HDAC3 was downregulated using small interfering RNA. Analysis of the expression of nuclear transcription factor-kappa B p65 (NF-κB p65) and molecules of its downstream signaling pathway were assessed and in lung tissue samples using the mouse model. Concentrations of tumor necrosis factor-α, interleukin-1β, high mobility group protein 1 (HMGB1), and interleukin-16 in supernatants and the bronchoalveolar lavage fluid were measured using enzyme-linked immunosorbent assay. A reporter gene assay measured HMGB1 activity, and NF-κB p65 and HMGB1 intracellular localization was determined by immunofluorescence detection on histological lung samples from Chr-treated mice. The protein interactions between HMGB1, HDAC3, and NF-κB p65 were tested by co-immunoprecipitation. Chr treatment relieved LPS-induced lung lesions. Chr also enhanced superoxide dismutase levels in ALI mice. Chr reduced the LPS-induced protein expression of NF-κB and its related pathway molecules in both and models. Moreover, Chr downregulated LPS-enhanced HMGB1 expression, acetylation, and nuclear nucleocytoplasmic translocation. However, HDAC3 knockdown substantially reduced Chr-mediated HDAC3/NF-κB expression. Furthermore, Chr enhanced HMGB1/HDAC3/NF-κB p65 complex interaction, whereas HDAC3 knockdown reduced Chr-mediated HMGB1/HDAC3/NF-κB p65 formation. This study showed that the protective effects induced by Chr were associated with the regulation of the HMGB1/NF-κB pathway via HDAC3.
大黄酚（Chr）是从……中分离出的主要单体。本研究旨在确定Chr对脂多糖（LPS）引发的急性肺损伤（ALI）的潜在抗炎和细胞保护作用。我们使用ALI小鼠模型并构建炎症巨噬细胞模型来确定Chr介导的活性所涉及的细胞机制。为了观察组蛋白去乙酰化酶3（HDAC3）在消除炎症作用中的关键作用，使用小干扰RNA下调HDAC3。使用小鼠模型评估肺组织样本中核转录因子-κB p65（NF-κB p65）及其下游信号通路分子的表达。使用酶联免疫吸附测定法测量上清液和支气管肺泡灌洗液中肿瘤坏死因子-α、白细胞介素-1β、高迁移率族蛋白1（HMGB1）和白细胞介素-16的浓度。报告基因测定法测量HMGB1活性，并通过对Chr处理小鼠的肺组织学样本进行免疫荧光检测来确定NF-κB p65和HMGB1的细胞内定位。通过共免疫沉淀测试HMGB1、HDAC3和NF-κB p65之间的蛋白质相互作用。Chr处理减轻了LPS诱导的肺部病变。Chr还提高了ALI小鼠中超氧化物歧化酶水平。Chr在细胞和小鼠模型中均降低了LPS诱导的NF-κB及其相关通路分子的蛋白表达。此外，Chr下调了LPS增强的HMGB1表达、乙酰化和核质转位。然而，HDAC3敲低显著降低了Chr介导的HDAC3/NF-κB表达。此外，Chr增强了HMGB1/HDAC3/NF-κB p65复合物相互作用，而HDAC3敲低减少了Chr介导的HMGB1/HDAC3/NF-κB p65形成。本研究表明，Chr诱导的保护作用与通过HDAC3调节HMGB1/NF-κB途径有关。
