Arana-Argáez Víctor Ermilo, Chan-Zapata Ivan, Canul-Canche Jaqueline, Fernández-Martín Karla, Martín-Quintal Zhelmy, Torres-Romero Julio Cesar, Coral-Martínez Tania Isolina, Lara-Riegos Julio Cesar, Ramírez-Camacho Mario Alberto
Laboratorio de Farmacología, Facultad de Química, Universidad Autónoma de Yucatán.
Laboratorio de Química Farmacéutica, Facultad de Química, Universidad Autónoma de, Yucatán.
Afr J Tradit Complement Altern Med. 2016 Nov 23;14(1):179-186. doi: 10.21010/ajtcam.v14i1.20. eCollection 2017.
The aim of this work was to evaluate the immunomodulatory effect of the methanol extract (MeOH) from leaves on the MΦs functions.
Peritoneal murine MΦs isolated from Balb/c mice were treated with the MeOH extract and stimulated with LPS. The effect on the phagocytosis was evaluated by flow cytometry assay. The nitric oxide (NO) and hydrogen peroxide (HO) production was measured by the Griess reagent and phenol red reaction, respectively. Levels of IL-6 and TNF-α was measured using an ELISA kit. Viability of MΦs and Vero cells was determined by the MTT method.
The MeOH extract of leaves inhibited significantly the phagocytosis, and decreased IL-6 and TNF-α production as well as NO and HO released by the MΦs, in a concentration-dependent manner. In addition, MeOH extract of showed low cytotoxicity effect against the cells.
These results suggest that MeOH extract of leaves has an immunosuppressive effect on murine MΦs, without effects on cell viability. GC-MS chromatogram analysis of MeOH extract showed that lupeol acetate and alpha-amyrin acetate are the principal compounds.
本研究旨在评估叶甲醇提取物（MeOH）对巨噬细胞（MΦs）功能的免疫调节作用。
从Balb/c小鼠分离出的腹膜巨噬细胞用MeOH提取物处理并用脂多糖（LPS）刺激。通过流式细胞术检测评估对吞噬作用的影响。分别用格里斯试剂和酚红反应测定一氧化氮（NO）和过氧化氢（HO）的产生量。使用酶联免疫吸附测定试剂盒（ELISA试剂盒）测量白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的水平。通过MTT法测定巨噬细胞和非洲绿猴肾细胞（Vero细胞）的活力。
叶的MeOH提取物以浓度依赖性方式显著抑制吞噬作用，并降低巨噬细胞释放的IL-6和TNF-α的产生量以及NO和HO的释放量。此外，叶的MeOH提取物对细胞显示出低细胞毒性作用。
这些结果表明叶的MeOH提取物对小鼠巨噬细胞具有免疫抑制作用，而对细胞活力无影响。MeOH提取物的气相色谱 - 质谱联用（GC-MS）色谱分析表明，乙酸羽扇豆醇酯和乙酸α-香树脂醇酯是主要化合物。
