Lupinus mutabilis Sweet(Tarwi)γ-凝集素调节受秘鲁利什曼原虫(V. Peruviana)感染的人类单核细胞中 TNF-α 和 IL-1α 的转录表达

R. Arone-Farfán, L. Alzamora-Gonzales, L.A.R. Carnero, E. Colona-Vallejos, E. Escobar-Guzmán, F. Camarena-Mayta, A. Huaringa-Joaquín
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摘要

背景:γ-凝集素是一种血凝凝集素,存在于安第斯豆科植物 Lupinus mutabilis 的种子中。秘鲁利什曼原虫(Viannia)是皮肤利什曼病的病原体,该病在秘鲁流行。利什曼病是一种被忽视的疾病,缺乏有效和无毒的治疗方法。本研究的目的是评估在感染 L. (V.) peruviana 之前,来自 L. mutabilis Patón Grande 生态型(CoPG)的 γ-凝集素对人类单核细胞培养的影响,并确定刺激细胞免疫的分子的产生情况。方法:通过色谱法从 L. mutabilis 种子的正己烷提取物中分离出 CoPG。通过密度梯度离心获得单核细胞。通过 MTT 试验确定 CoPG 对单核细胞活力的影响。为了评估超氧化物和一氧化氮的产生,先用 CoPG 对单核细胞进行预处理,然后用 L. (V.) peruviana 感染。通过 RT-PCR 法测定经 CoPG 处理和感染寄生虫的单核细胞中 TNF-α 和 IL-1α 的转录表达。结果显示在整个分离过程中,CoPG 始终保持其血凝活性。纯化的蛋白质分子量为 47 KDa。此外,20 µg mL-1 浓度的 CoPG 能诱导处理过的单核细胞产生超氧化物。这种浓度的 CoPG 还能促进受感染单核细胞中 IL-1α 和 TNF-α 的过度表达。本研究表明,CoPG 具有新的体外免疫调节作用。从L. mutabilis中发现生物活性蛋白可能会为这种安第斯植物带来广泛的新应用。
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Lupinus mutabilis Sweet (Tarwi) γ-conglutin Modulates TNF-α and IL-1α Transcriptional Expression in Human Monocytes Infected with Leishmania (V.) peruviana
Background: γ-Conglutin is a hemagglutinating lectin found in the seeds of the Andean legume Lupinus mutabilis. Leishmania (Viannia) peruviana is the etiological agent of cutaneous leishmaniasis, which is endemic in Peru. Leishmaniasis is a neglected disease that lacks an effective and non-toxic treatment. The aim of this study was to evaluate the effect of γ-conglutin from L. mutabilis Patón Grande ecotype (CoPG) on human monocyte cultures before infection with L. (V.) peruviana and to determine the production of molecules that stimulate cellular immunity. Methods: CoPG was isolated by chromatography from a hexane extract of L. mutabilis seeds. Monocytes were obtained by density gradient centrifugation. The effect of CoPG on their viability was determined by the MTT assay. To assess superoxide and nitric oxide production, monocyte cultures were pre-treated with CoPG and then infected with L. (V.) peruviana. Transcriptional expression of TNF-α and IL-1α in CoPG-treated and parasite-infected monocytes was determined by RT-PCR. Result: CoPG retained its hemagglutinating activity throughout the isolation process. The purified protein had a molecular weight of 47 KDa. Besides, CoPG at a 20 µg mL-1 induced the production of superoxide on the treated monocytes. This CoPG concentration also promoted the overexpression of IL-1α and TNF-α on the infected monocytes. The present study showed that CoPG has a novel in vitro immunomodulatory effect. The discovery of bioactive proteins from L. mutabilis may uncover a wide range of new applications for this Andean plant.
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