Center for Proteomic and Biochemical Analyses, Post-Graduate Program in Genomic Sciences and Biotechnology, Catholic University of Brasilia, SGAN Quadra 916, Modulo B, Av. W5, 70.790-160 Brasília, DF, Brazil.
Peptides. 2010 Aug;31(8):1426-33. doi: 10.1016/j.peptides.2010.05.003. Epub 2010 May 16.
Plants have contributed over the years to the discovery of various pharmacological products. Amongst the enormous diversity of herbs with remarkable medicinal use and further pharmacological potential, here in this report we evaluated pulp extracts from Eugenia dysenterica fruits and further identified the active principle involved in such laxative activity in rats. For protein isolation, fruits were macerated with an extraction solution following precipitation with (NH(4))(2)SO(4) (100%). After dialysis, the peptide was applied onto a reversed-phase semi-preparative HPLC column, and the major fraction was eluted with 26% and 66% acetonitrile. The evaluation of molecular masses by MALDI-TOF and Tris/Tricine SDS-PAGE of HPLC fractions showed the presence of a major peptide with approximately 7 kDa. The N-terminal amino acid peptide sequence was determined and showed no similarity to other proteins deposited in the Data Bank. Peptide from E. dysenterica was able to enhance rats' intestinal motility by approximately 20.8%, probably being responsible for laxative activity. Moreover, these proteins were non-toxic to mammals, as observed in histopathology and hemolytic analyses. In conclusion, results here reported indicate that, in the near future, proteins synthesized by E. dysenterica fruits could be utilized in the development of novel biotechnological pharmaceutics with laxative properties for use in chronic constipation and irritable bowel syndrome treatment.
多年来,植物为发现各种药理产品做出了贡献。在具有显著药用价值和进一步药理潜力的大量草药多样性中,在此报告中,我们评估了醉果果实的果肉提取物,并进一步确定了其在大鼠中具有通便活性的活性成分。为了进行蛋白质分离,将果实用提取溶液浸渍,然后用(NH(4))(2)SO(4)(100%)沉淀。透析后,肽被应用于反相半制备 HPLC 柱,主要馏分用 26%和 66%乙腈洗脱。通过 MALDI-TOF 和 HPLC 馏分的 Tris/Tricine SDS-PAGE 评估分子量,显示存在约 7 kDa 的主要肽。确定了 N 末端氨基酸肽序列,与数据库中存储的其他蛋白质没有相似性。来自醉果的肽能够使大鼠的肠道蠕动增加约 20.8%,可能是通便活性的原因。此外,这些蛋白质对哺乳动物没有毒性,如组织病理学和溶血分析所观察到的那样。总之,本报告的结果表明,在不久的将来,醉果果实合成的蛋白质可用于开发具有通便特性的新型生物技术药剂,用于治疗慢性便秘和肠易激综合征。