Bastos Andréa Patrícia da Silva Pomposo, Cardoso Patrícia Gomes, Santos Ítalo Augusto Férrer Melo, Trento Marcus Vinicius Cardoso, Porto Laura Cristina Jardim, Marcussi Silvana
Nutrition Department (Departamento de Nutrição), Universidade Federal de Lavras (UFLA), Lavras, MG, 37200-900, Brazil.
Biology Department (Departamento de Biologia), BIOGEN Laboratory (Laboratório BIOGEN), Universidade Federal de Lavras (UFLA), Lavras, MG, 37200-900, Brazil.
Curr Microbiol. 2020 Nov;77(11):3603-3611. doi: 10.1007/s00284-020-02170-5. Epub 2020 Aug 31.
In the present work, ethyl acetate extracts, consisting of non-volatile compounds, from the culture of endophytic fungi isolated from coffee plants, Induratia coffeana and Induratia yucatanensis, were prospected in enzyme modulation tests that act in human hemostasis. Dry extracts of the fungi were diluted in dimethyl sulfoxide p.a. 99.9% (DMSO), and then tested. Bothrops atrox venom was used as an enzyme source and tool to induce the activities. Prior to the evaluation of the activities, incubations of the extracts with the venom were performed in the proportions 1: 0.01, 1: 0.25, 1: 0.5, and 1: 1 (venom: extract; mass: mass). The extracts of all fungi promoted a significant increase in the clotting time induced by the venom, which was even longer when the extracts were previously incubated with the citrated plasma. The activity of phospholipases A did not significantly change when evaluated in the presence of fungal extracts. However, the evaluated extracts inhibited proteases by 73% and 30% in the thrombolytic and caseinolytic tests, respectively. In addition, the extracts did not induce cytotoxicity on human erythrocytes when evaluated in the absence of the venom. Thus, it is possible to suggest the presence of specific interactions between molecules present in extracts of Induratia spp. and venom proteases, highlighting non-volatile metabolites as promising sources of compounds of medical and scientific interest.
在本研究中，从咖啡植物Induratia coffeana和Induratia yucatanensis中分离出的内生真菌培养物中的由非挥发性化合物组成的乙酸乙酯提取物，在作用于人体止血的酶调节试验中进行了探索。将真菌的干提取物用99.9%的分析纯二甲基亚砜（DMSO）稀释，然后进行测试。以矛头蝮蛇毒作为酶源和诱导活性的工具。在评估活性之前，将提取物与蛇毒按1:0.01、1:0.25、1:0.5和1:1（蛇毒:提取物；质量:质量）的比例进行孵育。所有真菌的提取物均使蛇毒诱导的凝血时间显著延长，当提取物预先与枸橼酸盐血浆孵育时，凝血时间甚至更长。在真菌提取物存在的情况下评估时，磷脂酶A的活性没有显著变化。然而，在溶栓和酪蛋白溶解试验中，所评估的提取物分别抑制蛋白酶73%和30%。此外，在没有蛇毒的情况下评估时，提取物对人红细胞没有诱导细胞毒性。因此，可以推测Induratia spp.提取物中的分子与蛇毒蛋白酶之间存在特异性相互作用，突出了非挥发性代谢物作为具有医学和科学意义的化合物的有前景来源。
