Li Ping, Liang Qiao-Li, Cui Xiao-Dong, Li Jun, Zou Nuo-Shu, Wu Qi-Nan, Duan Jin-Ao
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China.
J Ethnopharmacol. 2014 Dec 2;158 Pt A:331-7. doi: 10.1016/j.jep.2014.10.032. Epub 2014 Oct 31.
Scirpus yagara Ohwi is a perennial, aquatic plant, whose dry tubers have long been used as Traditional Chinese Medicine (TCM) "Sanleng" for the treatment of postpartum abdominal pain, hyperemesis gravidarum, amenorrhea, dyspepsia and several inflammatory related diseases. Although it is known to have anti-inflammatory activities, its mechanism of action on lipopolysaccharide (LPS)-induced inflammation has not yet been identified in detail.This study was designed to investigate the anti-inflammatory activity of the active fraction (AF) from the tuber of Scirpusyagara both in vitro and in vivo.
RAW264.7 macrophage was incubated for 16h with 1µg/ml of LPS in absence or presence of AF (0, 10, 50 and 100µg/ml) and the secretions of tumor necrosis-alpha (TNF-α) and interleukin-6 (IL-6) in the medium were determined by using an enzyme-linked immunosorbent assay (ELISA). In the in vivo study, mice were orally administrated with AF (50 and 300mg/kg) for three days consecutively. 1h after the last AF administration, the mice were intraperitoneally injected with LPS (15mg/kg), and the life span of LPS-challenged mice were determined. Furthermore, the levels of pro-inflammatory cytokines TNF-α and IL-6 in the serum, lung and liver were measured using ELISA kit, and histological change in lungs was examined by light microscopy. Additionally, the components of AF were analyzed by high performance liquid chromatography (HPLC) using a C18 column.
AF significantly decreased TNF-α and IL-6 production induced by LPS in RAW264.7 macrophage. In LPS-induced mouse endotoxin shock model, AF pre-treatment significantly improved the survival rate of mice. And LPS-induced increases of pro-inflammatory cytokines TNF-α and IL-6 in the serum, lung and liver were markedly suppressed by AF. Moreover, the histopathological examination indicated that AF could significantly attenuate lung tissues injury in endotoxemic mice. In addition, eight compounds (protocatechuic acid, vanillic acid, p-coumaric acid, ferulic acid, methyl-3,6-dihydroxy-2-[2-(2-hydroxyphenyl)-ethynyl] benzoate, sciryagarol I, sparstolonin B, SanLeng diphenyllactone) of AF were quantified by HPLC analysis.
These results suggested that AF protected mice against LPS-induced lethality by inhibiting the production of multiple cytokines and organ dysfunction. Thus AF may prove beneficial in the prevention and treatment of endotoxin shock.
三棱是一种多年生水生植物，其干燥块茎长期以来一直被用作传统中药“三棱”，用于治疗产后腹痛、妊娠剧吐、闭经、消化不良以及几种炎症相关疾病。尽管已知其具有抗炎活性，但其对脂多糖（LPS）诱导的炎症的作用机制尚未详细阐明。本研究旨在研究三棱块茎活性成分（AF）在体外和体内的抗炎活性。
将RAW264.7巨噬细胞与1μg/ml LPS在不存在或存在AF（0、10、50和100μg/ml）的情况下孵育16小时，并使用酶联免疫吸附测定（ELISA）法测定培养基中肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的分泌量。在体内研究中，小鼠连续三天口服给予AF（50和300mg/kg）。最后一次给予AF 1小时后，小鼠腹腔注射LPS（15mg/kg），并测定LPS攻击小鼠的存活时间。此外，使用ELISA试剂盒测定血清、肺和肝脏中促炎细胞因子TNF-α和IL-6的水平，并通过光学显微镜检查肺组织的组织学变化。另外，使用C18柱通过高效液相色谱（HPLC）分析AF的成分。
AF显著降低LPS诱导的RAW264.7巨噬细胞中TNF-α和IL-6的产生。在LPS诱导的小鼠内毒素休克模型中，AF预处理显著提高了小鼠的存活率。并且AF显著抑制了LPS诱导的血清、肺和肝脏中促炎细胞因子TNF-α和IL-6的增加。此外，组织病理学检查表明AF可显著减轻内毒素血症小鼠的肺组织损伤。另外，通过HPLC分析对AF中的8种化合物（原儿茶酸、香草酸、对香豆酸、阿魏酸、3,6-二羟基-2-[2-(2-羟基苯基)-乙炔基]苯甲酸甲酯、三棱螺内酯I、水蓼二醛、三棱二苯内酯）进行了定量。
这些结果表明，AF通过抑制多种细胞因子的产生和器官功能障碍来保护小鼠免受LPS诱导的致死性。因此，AF可能在预防和治疗内毒素休克方面证明是有益的。
