Programa Multi-institucional de Pós-graduação em Biotecnologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil; Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil.
Laboratório de Imunoquímica, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas, Manaus, AM, Brazil.
J Ethnopharmacol. 2017 Jan 20;196:168-177. doi: 10.1016/j.jep.2016.12.031. Epub 2016 Dec 18.
The effectiveness of aqueous extract of Bellucia dichotoma Cogn. (Melastomataceae) specimems collected in Santarém, PA, against some biological activities of Bothrops atrox venom (BaV) has been scientifically proven. Here, we analyzed the components and assessed the anti-snakebite potential of aqueous extracts of bark of B. dichotoma collected in Manaus, AM, (AEBd-MAO) and Santarém, PA, (AEBd-STM), both in Brazil.
The phytochemical profiles of the aqueous extracts were identified using thin layer chromatography (TLC), and the concentrations of phenolics were determined by colorimetric assay. The inhibitory potential of the extracts was tested against the phospholipase A, coagulant and gelatinolytic activities of BaV in vitro and its defibrinating and edema-inducing activities in vivo. Interaction between BaV and the extracts was investigated using SDS-Page electrophoresis and Western blotting. Extract cytotoxicity and antioxidant potential were assessed using the human fibroblast cell line MRC-5 and the DPPH assay in cell culture, respectively.
While there was no difference between the phytochemical profiles of the extracts, AEBd-MAO had higher concentrations of total phenolics, total tannins and hydrolysable tannins. The extracts inhibited 100% of the phospholipase and coagulant activity of BaV when pre-incubated. Without pre-incubation, however, there was no reduction in phospholipase activity, although significant inhibition of coagulant activity was observed. In the doses used in folk medicine, without pre-incubation, both extracts inhibited 100% of the coagulant activity of BaV. In vivo, the extracts were unable to inhibit the defibrinating activity of the venom but were effective in inhibiting its edema-inducing activity. In the profiles of the extracts pre-incubated with BaV, not all the protein bands revealed by SDS-PAGE and Western blot were observed. Both extracts had a high antioxidant potential and neither had a cytotoxic effect.
Although the concentrations of phenolics in each extract were different, the anti-snakebite potential was similar for the concentrations of extract tested. Our findings are of importance for the quality control of this raw material, which, once tested in accordance with Brazilian National Health Surveillance Agency recommendations, may be suitable for use as a phytomedicine to complement treatment of the local effects induced by Bothrops venoms.
已从科学上证明,产自帕拉州圣塔伦的 Bellucia dichotoma Cogn.(野牡丹科)水提物标本对矛头蝮蛇(Bothrops atrox venom,BaV)的一些生物活性具有疗效。在此,我们分析了产自亚马孙州马瑙斯(AEBd-MAO)和帕拉州圣塔伦(AEBd-STM)的 Bellucia dichotoma 树皮的水提物成分,并评估了其抗蛇毒的潜力。
采用薄层色谱(TLC)鉴定水提物的植物化学成分,并通过比色法测定其酚类浓度。在体外测试了提取物对 BaV 的磷脂酶、凝血和明胶酶活性的抑制潜力,并在体内测试了其抗纤溶和致水肿活性。采用 SDS-Page 电泳和 Western blot 研究了 BaV 与提取物的相互作用。通过人成纤维细胞系 MRC-5 和 DPPH 测定法分别在细胞培养中评估提取物的细胞毒性和抗氧化潜力。
尽管提取物的植物化学成分图谱没有差异,但 AEBd-MAO 的总酚、总单宁和可水解单宁的浓度更高。提取物在预孵育时可完全抑制 BaV 的 100%的磷脂酶和凝血活性。然而,没有预孵育时,虽然观察到凝血活性显著抑制,但磷脂酶活性没有降低。在民间医学中使用的剂量下,即使没有预孵育,两种提取物也可完全抑制 BaV 的凝血活性。在体内,提取物不能抑制蛇毒的纤溶活性,但能有效抑制其致水肿活性。在与 BaV 预孵育的提取物图谱中,SDS-Page 和 Western blot 揭示的并非所有蛋白带均可见。两种提取物均具有较高的抗氧化潜力,且均无细胞毒性。
尽管每个提取物的酚类浓度不同,但在所测试的提取物浓度下,抗蛇毒的潜力相似。我们的研究结果对于该原料的质量控制具有重要意义,一旦按照巴西国家卫生监督局的建议进行测试,它可能适合作为一种植物药,用于补充治疗当地由矛头蝮蛇毒液引起的局部影响。