University of Münster, Institute of Pharmaceutical Biology and Phytochemistry, Corrensstrasse 48, D-48149 Münster, Germany; Centro de Referencia para Lactobacilos (CERELA)-CONICET, Chacabuco 145, Tucumán, San Miguel de Tucumán 4000, Argentina; INQUINOA-CONICET Instituto de Química Orgánica, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, S. M. de Tucumán T4000INI, Argentina.
University of Münster, Institute of Pharmaceutical Biology and Phytochemistry, Corrensstrasse 48, D-48149 Münster, Germany.
J Ethnopharmacol. 2014 Sep 11;155(2):1125-33. doi: 10.1016/j.jep.2014.06.039. Epub 2014 Jun 30.
ETHNOPHARMACOLOGICAL RELEVANCE: The aerial parts of Lippia integrifolia (Gris.) Hieronymus (Verbenaceae), known as incayuyo, are used by the peasant population of Argentina for treatment of diseases related to a gastrointestinal system, mainly for inflammation of the stomach and have also been included into the Argentina Food Code. This study aimed to investigate the phytochemical profile of hydrophilic extracts from the herbal material by LC-MS and to evaluate potential pharmacological mechanisms rationalizing the traditional use of incayuyo aqueous extracts potential anti-inflammatory treatment of gastrointestinal disorders. MATERIALS AND METHODS: Phytochemical profiling: LC-MS of an aqueous decoction. Antiadhesive effects against Helicobacter pylori: in vitro FACS assay using FITC-labeled bacteria and AGS human stomach cells. Influence of extracts on stomach cells and RAW 264.7 macrophages: MTT viability assay and BrdU proliferation ELISA. Influence of extracts on IL-6 and IL-8 secretion from stomach cells was quantitated by ELISA after infection of the cells with Helicobacter pylori. Influence of test extracts on macrophages: phagocytosis of FITC-labeled Zymosan particles and NO production. Antioxidative capacity: DPPH assay and O2-induced caroten oxidation. RESULTS: LC-MS profiling indicated the presence of compounds 1-20 with flavonoid hexosides, phenylethanoides (acteoside, isoacteoside) and sesquiterpenes [(epi)lippidulcine, peroxylippidulcine] in the decoction extract and subfraction PhF. The extract exhibits strong in vitro antioxidative capacity and inhibited adhesion of Helicobacter pylori to stomach cells up to 40%, while an EtOH-soluble fraction showed inhibition rates of up to 60%. Decoction increased the cellular viability of AGS cells significantly at >10 μg/mL, while proliferation rate was not influenced. Helicobacter pylori induced IL-8 secretion was significantly reduced by coincubation of AGS cells with the extracts. Aqueous extracts stimulated phagocytosis rate of macrophages and inhibited the LPS-induced NO-secretion. CONCLUSIONS: The traditional use of aqueous extracts from Lippia integrifolia for gastric inflammation seems to be rationalized: besides anti-inflammatory effects on stomach cells antiadhesive properties of the extracts against the main bacterial inductor of gastritis Helicobacter pylori were identified. Additional effects for stimulation of innate immunity and potential radical scavenging effects may additionally contribute to the activity of the extracts.
民族药理学相关性：唇形科 Lippia integrifolia（Gris.）Hieronymus 的地上部分（称为 incayuyo），被阿根廷农民用于治疗与胃肠道系统相关的疾病，主要用于治疗胃炎，并且已被纳入阿根廷食品法典。本研究旨在通过 LC-MS 分析亲水提取物的植物化学特征，并评估潜在的药理机制，以合理化 incayuyo 水提物用于治疗胃肠道疾病的传统用途，这种水提物具有抗炎作用。
材料和方法：植物化学特征分析：水提物的 LC-MS 分析。抗粘附作用：使用 FITC 标记的细菌和 AGS 人胃细胞的体外 FACS 分析。提取物对胃细胞和 RAW 264.7 巨噬细胞的影响：MTT 细胞活力测定和 BrdU 增殖 ELISA。通过用幽门螺杆菌感染细胞后定量测定 ELISA 上清液中细胞因子 IL-6 和 IL-8 的分泌来评估提取物对细胞因子的影响。提取物对巨噬细胞的影响：用 FITC 标记的 Zymosan 颗粒吞噬和 NO 产生。抗氧化能力：DPPH 测定法和 O2 诱导的类胡萝卜素氧化。
结果：LC-MS 分析表明，在水提物和 PhF 亚组分中存在化合物 1-20，包括黄酮糖苷、苯乙醇（acteoside、isoacteoside）和倍半萜类化合物[(epi)lippidulcine、peroxylippidulcine]。提取物具有较强的体外抗氧化能力，可抑制幽门螺杆菌与胃细胞的粘附，最高可达 40%，而 EtOH 可溶部分的抑制率最高可达 60%。水提物在>10μg/mL 时可显著增加 AGS 细胞的细胞活力，而对增殖率无影响。与提取物共培养可显著降低幽门螺杆菌诱导的 AGS 细胞中 IL-8 的分泌。水提物刺激巨噬细胞吞噬率，并抑制 LPS 诱导的 NO 分泌。
结论：传统上使用 Lippia integrifolia 的水提物治疗胃炎似乎是合理的：除了对胃细胞的抗炎作用外，还鉴定出提取物对胃炎的主要细菌诱导物幽门螺杆菌的抗粘附特性。此外，刺激先天免疫和潜在的自由基清除作用的作用可能会进一步增强提取物的活性。
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