Laboratorio de Investigaciones Bioquímicas de La Facultad de Medicina (LIBIM), Instituto de Química Básica y Aplicada Del NEA, (IQUIBA NEA- UNNE- CONICET), Facultad de Medicina, Universidad Nacional Del Nordeste, 3400, Corrientes, Argentina; Laboratorio de Productos Naturales Prof. Armando Ricciardi (LabProdNat), Instituto de Química Básica y Aplicada Del NEA, (IQUIBA NEA- UNNE- CONICET), Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional Del Nordeste, 3400, Corrientes, Argentina.
Laboratorio de Investigaciones Bioquímicas de La Facultad de Medicina (LIBIM), Instituto de Química Básica y Aplicada Del NEA, (IQUIBA NEA- UNNE- CONICET), Facultad de Medicina, Universidad Nacional Del Nordeste, 3400, Corrientes, Argentina.
J Ethnopharmacol. 2021 May 23;272:113937. doi: 10.1016/j.jep.2021.113937. Epub 2021 Feb 18.
Nectandra angustifolia belongs to the Lauraceae family and it is widely known in phytomedicine by local inhabitants of South America against various maladies. It is popularly used for the treatment of different types of inflammatory processes, like rheumatism, arthritis and its associated pain.
To characterize the phytochemicals in an ethanolic extract of Nectandra angustifolia and to evaluate the total antioxidant content and its anti-inflammatory effect with multiparametric analyses through in vitro assays and an in vivo model.
Leaves and stems of Nectandra angustifolia were air-dried and an ethanolic extract (NaE) was further obtained. Total phenolic, flavonoid and tannin content were determined and the antioxidant activity was addressed by DPPH and FRAP assays. NaE was first analyzed by HPLC and then two tests were carried out as screening assays for anti-inflammatory activities: red blood cell membrane stabilization and protein denaturation. The non-cytotoxic concentration of NaE was determined for in vitro biological assays using RAW 264.7 (murine macrophages) cell cultures through cell counting with Trypan-blue and XTT assay. Subsequently, the cell cycle of RAW 264.7 cells exposed for 24 h to NaE was analyzed. Additionally, the anti-inflammatory capacity of NaE was evaluated by RT-qPCR of pro-inflammatory cytokines. Furthermore, NF-κB translocation was observed by confocal microscopy at different times. Finally, formalin-induced mice paw inflammation was used as an in vivo model.
The chromatographic profile of NaE showed peaks compatible with flavonoids content. NaE exhibited better membrane stabilization effect on HRBC and protection of BSA denaturation than the standard drug (diclofenac). NaE diminished mRNA levels of pro-inflammatory cytokines when added 1-h prior LPS stimulation. Moreover, NaE prevented the translocation of NF-κB to the nucleus and in formalin-induced mice paw inflammation, reduced the edema and the stimulus of inflammatory phase.
This study shows for the first time, that Nectandra angustifolia ethanolic extract has a high content of flavonoids and that possess antioxidant and anti-inflammatory biological properties as demonstrated by multiparametric analyses from in vitro assays and an in vivo model of inflammation.
荷南杜拉 angustifolia 属于樟科，在南美洲，当地居民广泛应用它治疗各种疾病。在植物药中，它常用于治疗各种炎症过程，如风湿、关节炎及其相关疼痛。
研究荷南杜拉 angustifolia 的乙醇提取物中的植物化学成分，并通过体外试验和体内炎症模型的多参数分析，评估其总抗氧化含量和抗炎作用。
荷南杜拉 angustifolia 的叶子和茎被风干，然后得到乙醇提取物（NaE）。测定总酚、类黄酮和单宁的含量，并通过 DPPH 和 FRAP 测定法评估抗氧化活性。NaE 首先通过 HPLC 进行分析，然后进行两种筛选抗炎活性的试验：红细胞膜稳定性和蛋白质变性。通过使用台盼蓝和 XTT 测定法对 RAW 264.7（鼠巨噬细胞）细胞培养物进行细胞计数，确定 NaE 的非细胞毒性浓度，用于体外生物学试验。随后，分析暴露于 NaE 24 小时的 RAW 264.7 细胞的细胞周期。此外，通过 RT-qPCR 分析 NaE 对促炎细胞因子的抗炎能力。此外，通过共聚焦显微镜观察不同时间 NF-κB 的转位。最后，使用甲醛诱导的小鼠爪炎症作为体内模型。
NaE 的色谱图显示与类黄酮含量相符的峰。与标准药物（双氯芬酸）相比，NaE 对 HRBC 具有更好的膜稳定作用，对 BSA 变性的保护作用更强。当在 LPS 刺激前 1 小时加入 NaE 时，NaE 降低了促炎细胞因子的 mRNA 水平。此外，NaE 可防止 NF-κB 向细胞核转位，并减轻甲醛诱导的小鼠爪炎症的水肿和炎症期刺激。
本研究首次表明，荷南杜拉 angustifolia 乙醇提取物具有较高的类黄酮含量，并具有抗氧化和抗炎生物特性，这通过体外试验和体内炎症模型的多参数分析得到证明。
