Department of Pharmaceutical Technology, Centre for Excellence in Nanobio Translational Research, Bharathidasan Institute of Technology, Anna University, Tiruchirappalli, 620024, Tamil Nadu, India.
Department of Pharmaceutical Technology, Centre for Excellence in Nanobio Translational Research, Bharathidasan Institute of Technology, Anna University, Tiruchirappalli, 620024, Tamil Nadu, India; National Facility for Drug Development (NFDD) for Academia, Pharmaceutical and Allied Industries, Bharathidasan Institute of Technology, Anna University, Tiruchirappalli, 620024, Tamil Nadu, India.
J Ethnopharmacol. 2020 Jun 12;255:112369. doi: 10.1016/j.jep.2019.112369. Epub 2019 Nov 1.
Myxopyrum serratulum A. W. Hill. (Oleaceae) is a traditionally used Indian medicinal plant for the treatment of cough, asthma and many other inflammatory diseases.
In this study, the protective effects of M. serratulum on airway inflammation was investigated in ovalbumin (OVA)-induced murine model of allergic asthma and lipopolysaccharide (LPS)-stimulated inflammation in RAW 264.7 murine macrophages, and the possible mechanisms were elucidated.
The phytochemicals present in the methanolic leaf extract of M. serratulum (MEMS) were identified by reverse phase high performance liquid chromatography (RP-HPLC) analysis. In vitro anti-inflammatory activity of MEMS were evaluated by estimating the levels of nitric oxide (NO), reactive oxygen species (ROS) and cytokines (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17A, IFN-γ, TNF-α, G-CSF and GM-CSF) in LPS-stimulated RAW 264.7 macrophages. In vivo anti-asthmatic activity of MEMS was studied using OVA-induced murine model. Airway hyperresponsiveness (AHR), was measured; total and differential cell counts, eosinophil peroxidase (EPO), prostaglandin E2 (PGE2), NO, ROS, and cytokines (IL-4, IL-5 and IL-13), were estimated in bronchoalveolar lavage fluid (BALF). Serum total IgE level was measured; and the histopathological changes of lung tissues were observed. The expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in lung tissue homogenates were detected by Western blot.
The chromatographic analysis of MEMS identified the presence of gallic acid, protocatechuic acid, catechin, ellagic acid, rutin, p-coumaric acid, quercetin, naringenin and apigenin. MEMS (125 and 250 μg/mL) dose-dependently reduced the levels of NO, ROS and pro-inflammatory cytokines in LPS-stimulated RAW 264.7 macrophages. MEMS (200 and 400 mg/kg, p.o.) significantly (p < 0.05) alleviated AHR; number of inflammatory cells, EPO, PGE2, NO, ROS, and cytokines (IL-4, IL-5 and IL-13) in BALF; serum total IgE and the histopathological changes associated with lung inflammation. Western blot studies showed that MEMS substantially suppressed COX-2 and iNOS protein expressions in the lung tissues of OVA-sensitized/challenged mice.
The present study corroborates for the first time the ameliorative effects of MEMS on airway inflammation by reducing the levels of oxidative stress, pro-inflammatory cytokines and inhibiting COX-2, iNOS protein expressions, thereby validating the ethnopharmacological uses of M. serratulum.
Myxopyrum serratulum A. W. Hill.（木樨科）是一种传统的印度药用植物，用于治疗咳嗽、哮喘和许多其他炎症性疾病。
本研究旨在探讨 Myxopyrum serratulum 对卵清蛋白（OVA）诱导的过敏性哮喘小鼠模型气道炎症的保护作用，并在脂多糖（LPS）刺激的 RAW 264.7 小鼠巨噬细胞中阐明可能的机制。
采用反相高效液相色谱（RP-HPLC）分析鉴定 Myxopyrum serratulum 甲醇叶提取物（MEMS）中的植物化学物质。通过估计 LPS 刺激的 RAW 264.7 巨噬细胞中一氧化氮（NO）、活性氧（ROS）和细胞因子（IL-1α、IL-1β、IL-2、IL-4、IL-6、IL-10、IL-12、IL-17A、IFN-γ、TNF-α、G-CSF 和 GM-CSF）的水平，评估 MEMS 的体外抗炎活性。采用 OVA 诱导的小鼠模型研究 MEMS 的体内抗哮喘活性。测量气道高反应性（AHR）；测定支气管肺泡灌洗液（BALF）中的总细胞和分类细胞计数、嗜酸性粒细胞过氧化物酶（EPO）、前列腺素 E2（PGE2）、NO、ROS 和细胞因子（IL-4、IL-5 和 IL-13）。测定血清总 IgE 水平；观察肺组织的组织病理学变化。用 Western blot 检测肺组织匀浆中环氧化酶-2（COX-2）和诱导型一氧化氮合酶（iNOS）的表达。
MEMS 的色谱分析鉴定出存在没食子酸、原儿茶酸、儿茶素、鞣花酸、芦丁、对香豆酸、槲皮素、柚皮苷、橙皮苷和芹菜素。MEMS（125 和 250μg/mL）剂量依赖性地降低 LPS 刺激的 RAW 264.7 巨噬细胞中 NO、ROS 和促炎细胞因子的水平。MEMS（200 和 400mg/kg，po）显著（p<0.05）减轻 AHR；BALF 中炎症细胞、EPO、PGE2、NO、ROS 和细胞因子（IL-4、IL-5 和 IL-13）的数量；血清总 IgE 和与肺炎症相关的组织病理学变化。Western blot 研究表明，MEMS 可显著抑制 OVA 致敏/攻击小鼠肺组织中 COX-2 和 iNOS 蛋白的表达。
本研究首次证实了 MEMS 通过降低氧化应激、促炎细胞因子水平和抑制 COX-2、iNOS 蛋白表达来改善气道炎症，从而验证了 Myxopyrum serratulum 的民族药理学用途。
