Zheng Jing-Na, Zhuo Jian-Yi, Nie Juan, Liu Yan-Lu, Chen Bao-Yi, Wu Ai-Zhi, Li Yu-Cui
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Department of Pharmacy, The First Affiliated Hospital/School of Clinical Medicine of Guangdong Pharmaceutical University, Guangzhou, China.
Front Pharmacol. 2021 Jul 7;12:693983. doi: 10.3389/fphar.2021.693983. eCollection 2021.
Acute lung injury (ALI) is a complicated and severe lung disease, which is often characterized by acute inflammation. Poliumoside (POL), acteoside (ACT) and forsythiaside B (FTB) are phenylethanoid glycosides (PGs) with strong antioxidant, anti-inflammatory, and anti-apoptotic properties, which are extracted from (CK). The aim of this study was to investigate the protective effects of POL, ACT, and FTB against TNF-α-induced damage using an ALI cell model and explore their potential mechanisms. MTT method was used to measure cell viability. Flow cytometry was used for detecting the apoptosis rate. Reactive oxygen species (ROS) activity was determined using fluorescence microscope. The expression of mRNA in apoptosis-related genes (Caspase 3, Caspase 8, and Caspase 9) were tested by qPCR. The effects of POL, ACT, FTB on the activities of nuclear factor erythroid-2 related factor 2 (Nrf2), nuclear factor kappa-B (NF-κB) and the expression of their downstream genes were assessed by western blotting and RT-PCR in A549 cells. In the current study, POL, ACT, and FTB dose-dependently attenuated TNF-α-induced IL-1β, IL-6 and IL-8 production, cell apoptosis, the expression of apoptosis-related genes (Caspase 3, Caspase 8, and Caspase 9) and ROS activity. POL, ACT, and FTB not only increased in the mRNA levels of antioxidative enzymes NADPH quinone oxidoreductase (NQO1), glutamate cysteine ligase catalytic subunit (GCLC), heme oxygenase (HO-1), but also decreased the mRNA levels of IL-1β, IL-6 and IL-8. Furthermore, they upregulated the expression of Keap1 and enhanced the activation of Nrf2, while decreased the expression of phosphor-IκBα (-IκBα) and nuclear p65. In addition, no significant changes were observed in anti-inflammatory and antioxidant effects of POL, ACT, FTB following Nrf2 and NF-κB p65 knockdown. Our study revealed that POL, ACT, and FTB alleviated oxidative damage and lung inflammation of TNF-α-induced ALI cell model through regulating the Nrf2 and NF-κB pathways.
急性肺损伤（ALI）是一种复杂且严重的肺部疾病，常以急性炎症为特征。梓醇（POL）、毛蕊花糖苷（ACT）和连翘酯苷B（FTB）是从[此处原文缺失提取来源相关内容，推测可能是某种植物，用CK暂代]中提取的具有强抗氧化、抗炎和抗凋亡特性的苯乙醇苷（PGs）。本研究旨在利用ALI细胞模型研究POL、ACT和FTB对肿瘤坏死因子-α（TNF-α）诱导损伤的保护作用，并探索其潜在机制。采用MTT法检测细胞活力。运用流式细胞术检测凋亡率。使用荧光显微镜测定活性氧（ROS）活性。通过实时定量聚合酶链反应（qPCR）检测凋亡相关基因（半胱天冬酶3、半胱天冬酶8和半胱天冬酶9）的mRNA表达。通过蛋白质免疫印迹法（western blotting）和逆转录-聚合酶链反应（RT-PCR）评估POL、ACT、FTB对A549细胞中核因子红细胞2相关因子2（Nrf2）、核因子κB（NF-κB）活性及其下游基因表达的影响。在本研究中，POL、ACT和FTB呈剂量依赖性地减轻TNF-α诱导的白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和白细胞介素-8（IL-8）生成、细胞凋亡、凋亡相关基因（半胱天冬酶3、半胱天冬酶8和半胱天冬酶9）的表达以及ROS活性。POL、ACT和FTB不仅增加了抗氧化酶烟酰胺腺嘌呤二核苷酸磷酸醌氧化还原酶（NQO1）、谷氨酸半胱氨酸连接酶催化亚基（GCLC）、血红素加氧酶（HO-1）的mRNA水平，还降低了IL-1β、IL-6和IL-8的mRNA水平。此外，它们上调了 Kelch样环氧氯丙烷相关蛋白1（Keap1）的表达并增强了Nrf2的激活，同时降低了磷酸化IκBα（-IκBα）和核p65的表达。此外，在Nrf2和NF-κB p65基因敲低后，未观察到POL、ACT、FTB的抗炎和抗氧化作用有显著变化。我们的研究表明，POL、ACT和FTB通过调节Nrf2和NF-κB信号通路减轻了TNF-α诱导的ALI细胞模型的氧化损伤和肺部炎症。
