Santos Francianne Oliveira, de Lima Hélimar Gonçalves, de Souza Santos Nathália Silva, Serra Taiane Menezes, Uzeda Rosângela Soares, Reis Isabella Mary Alves, Botura Mariana Borges, Branco Alexsandro, Batatinha Maria José Moreira
Laboratório de Toxicologia, Hospital de Medicina Veterinária, Universidade Federal da Bahia, Brazil.
Laboratório de Fitoquímica, Universidade Estadual de Feira de Santana, Brazil.
Vet Parasitol. 2017 Oct 15;245:48-54. doi: 10.1016/j.vetpar.2017.08.007. Epub 2017 Aug 16.
This study aimed to evaluate the in vitro activity of D. insularis extracts and fractions against gastrointestinal nematodes of goats and its cytotoxicity on Vero cells. The egg hatch (EHT) and larval motility (LMT) tests were conducted to investigate the anthelmintic effects of the crude hydroethanolic (CH), ethyl acetate (EA), butanolic (BT) and residual hydroethanolic (RH) extracts. The elution of the active extract (EA) on column chromatography (SiO) using organic solvents furnished six fractions (FR1 to FR6), which were also tested. Cytotoxicity was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trypan Blue exclusion assays. All extracts, FR2 and FR3, inhibited egg hatching in a concentration-dependent manner. The EHT led to EC values (effective concentration 50%) of 0.64; 0.69; 0.77; 0.96; 0.27 and 0.65mg/mL for CH, EA, BT, RH, FR2 and FR3, respectively. However, the extracts exhibited low effect on the motility of L In the cytotoxicity evaluation (MTT assay), the IC (inhibitory concentration 50%) was 1.18 (EA), 1.65 (FR2) and 1.59mg/mL (FR3), which was relatively high (low toxicity) in comparison to the EC values in EHT, mainly for FR2. The chemical analyses of most active fractions (FR2) by Liquid Chromatography coupled to Mass Spectrometry (LC-MS) led the characterization of the flavones tricin and diosmetin. These results showed the high anthelmintic effect and low cytotoxicity of D. insularis and also that the flavones can be probably responsible for the nematocidal activity of this plant.
本研究旨在评估海岛龙葵提取物及其馏分对山羊胃肠道线虫的体外活性及其对Vero细胞的细胞毒性。进行了虫卵孵化(EHT)和幼虫运动(LMT)试验,以研究粗制氢乙醇提取物(CH)、乙酸乙酯提取物(EA)、丁醇提取物(BT)和残余氢乙醇提取物(RH)的驱虫效果。使用有机溶剂对活性提取物(EA)进行柱色谱(SiO)洗脱,得到六个馏分(FR1至FR6),并对其进行了测试。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)和台盼蓝排斥试验测定细胞毒性。所有提取物、FR2和FR3均以浓度依赖性方式抑制虫卵孵化。EHT试验中,CH、EA、BT、RH、FR2和FR3的EC值(有效浓度50%)分别为0.64;0.69;0.77;0.96;0.27和0.65mg/mL。然而,这些提取物对L的运动性影响较小。在细胞毒性评估(MTT试验)中,IC(抑制浓度50%)分别为1.18(EA)、1.65(FR2)和1.59mg/mL(FR3),与EHT试验中的EC值相比相对较高(低毒性),主要是FR2。通过液相色谱-质谱联用(LC-MS)对大多数活性馏分(FR2)进行化学分析,鉴定出黄酮类化合物小麦黄素和香叶木素。这些结果表明海岛龙葵具有较高的驱虫效果和较低的细胞毒性,并且黄酮类化合物可能是该植物杀线虫活性的原因。