Área de Farmacologia, Departamento de Ciências Básicas em Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso (UFMT), Cuiabá, MT, Brazil.
Área de Histologia e Biologia Celular, Departamento de Ciências Básicas em Saúde, Faculdade de Medicina, Universidade Federal de Mato Grosso (UFMT), Cuiabá, MT, Brazil.
J Ethnopharmacol. 2020 May 23;254:112707. doi: 10.1016/j.jep.2020.112707. Epub 2020 Feb 27.
ETHNOPHARMACOLOGICAL RELEVANCE: Piper umbellatum L. leaves, commonly found in the Amazon, Cerrado and Atlantic rain forest regions of Brazil, are widely used as a traditional medicine to treat gastrointestinal disorders and inflammation, among others diseases. Also, previous scientific studies demonstrated that P. umbellatum has gastroprotective and anti-inflammatory activity. AIM: To investigate the phytochemical profiles and the intestinal anti-inflammatory effect of the hydroethanolic extract of P. umbellatum (HEPu) leaf on ulcerative colitis in rats. MATERIALS AND METHODS: In this study, the chemical composition of HEPu was analyzed by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography coupled to mass spectrometry (LC/MS). Also, this work studied the effects of HEPu on ulcerative colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS, 30 mg/mL in 20% ethanol) by intrarectal administration in rats. Simultaneously, animals were pre-treated orally with HEPu (30, 100 and 300 mg/kg), mesalazine (500 mg/kg), or vehicle. At the end of the experimental period, clinical signs of ulcerative colitis were evaluated by determination of weight loss, gross appearance, ulcer area and histological changes. Reduced glutathione (GSH), lipoperoxides (MDA) and nitric oxide (NO) levels, and superoxide dismutase (SOD), myeloperoxidase (MPO) and catalase (CAT) activities were determined in colon tissues. Also, pro-inflammatory mediators such as tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL- 1β) were quantified by immunoassay on the surface of fluorescent-coded magnetic beads (Luminex MagPix System). RESULTS: GC-MS analysis showed the presence of 17 different phytochemical compounds in the HEPu. LC/MS analyses revealed the presence of compounds in HEPu as protocatechuic acid, ferulic acid, kaempferol, rosmarinic acid, apigenin and ursolic acid. Treatment with HEPu significantly ameliorated weight loss, macroscopic damage, ulcerated area and histopathological changes such as sub-mucosal edema, cell infiltration, ulceration and necrosis (p < 0.001). Furthermore, HEPu (30, 100, and 300 mg/kg, p.o) inhibited the levels of oxidative parameters, such as MPO (49%, 53%, and 62%, p < 0.001), NO (20%, 19%, 22%, p < 0.01), and MDA (75%, 83%, 70%, p < 0.001), whereas increased the antioxidant activities such as SOD (208%, 192%, 64%, p < 0.001), GSH (94%, 75%, 49%, p < 0.01), and CAT (92%, 69%, 108%, p < 0.01). The extract also inhibited the pro-inflammatory cytokines TNF-α (81%, 85%, 85%, p < 0.001) and IL-1β (95%, 79%, 89%, p < 0.001) levels. CONCLUSION: Together, these results revealed that P. umbellatum L. is a promising source of metabolites to be used in the treatment of inflammatory bowel disease.
民族药理学相关性：在亚马逊、塞拉多和大西洋雨林地区广泛发现的 Piper umbellatum L. 叶，被广泛用作传统药物来治疗胃肠道疾病和炎症等疾病。此外，先前的科学研究表明，Piper umbellatum 具有胃保护和抗炎活性。
目的：研究 Piper umbellatum（HEPu）叶水醇提取物对溃疡性结肠炎大鼠的肠道抗炎作用及其植物化学成分谱。
材料和方法：本研究通过气相色谱-质谱联用（GC-MS）和液相色谱-质谱联用（LC/MS）分析 HEPu 的化学成分。此外，本研究通过直肠内给予 2,4,6-三硝基苯磺酸（TNBS，20%乙醇中的 30mg/mL）诱导大鼠溃疡性结肠炎，研究 HEPu 的作用。同时，动物经口预先用 HEPu（30、100 和 300mg/kg）、美沙拉嗪（500mg/kg）或载体处理。在实验期末，通过测定体重减轻、大体外观、溃疡面积和组织学变化来评估溃疡性结肠炎的临床症状。通过测定结肠组织中的还原型谷胱甘肽（GSH）、脂质过氧化物（MDA）和一氧化氮（NO）水平以及超氧化物歧化酶（SOD）、髓过氧化物酶（MPO）和过氧化氢酶（CAT）活性来评估氧化应激参数。此外，通过荧光编码磁珠上的免疫测定法（Luminex MagPix 系统）定量测定促炎介质肿瘤坏死因子-α（TNF-α）和白细胞介素 1β（IL-1β）。
结果：GC-MS 分析显示 HEPu 中存在 17 种不同的植物化学化合物。LC/MS 分析显示 HEPu 中存在原儿茶酸、阿魏酸、山奈酚、迷迭香酸、芹菜素和熊果酸等化合物。HEPu 治疗显著改善了体重减轻、宏观损伤、溃疡面积和组织病理学变化，如黏膜下水肿、细胞浸润、溃疡和坏死（p<0.001）。此外，HEPu（30、100 和 300mg/kg，po）抑制了氧化应激参数水平，如 MPO（49%、53%和 62%，p<0.001）、NO（20%、19%和 22%，p<0.01）和 MDA（75%、83%和 70%，p<0.001），同时增加了抗氧化活性，如 SOD（208%、192%和 64%，p<0.001）、GSH（94%、75%和 49%，p<0.01）和 CAT（92%、69%和 108%，p<0.01）。该提取物还抑制了促炎细胞因子 TNF-α（81%、85%和 85%，p<0.001）和 IL-1β（95%、79%和 89%，p<0.001）水平。
结论：综上所述，这些结果表明 Piper umbellatum L. 是一种有前途的代谢产物来源，可用于治疗炎症性肠病。
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