Aguiar Jaciana S, Araújo Rosilma O, Do Desterro Rodrigues Maria, Sena Kêsia X F R, Batista André M, Guerra Maria M P, Oliveira Steno L, Tavares Josean F, Silva Marcelo S, Nascimento Silene C, Da Silva Teresinha Gonçalves
Antibiotics of Departament, Federal University of Pernambuco, 50670-901, Recife, PE, Brasil.
Andrology Laboratory, Veterinary Medicine Department, Rural Federal University of Pernambuco, 52171-900, Recife, PE, Brasil.
Int J Mol Sci. 2012;13(4):4124-4140. doi: 10.3390/ijms13044124. Epub 2012 Mar 29.
In the study, we have examined the antitumor and antimicrobial activities of the methanol extract, the fractions, a fraction of total alkaloids and two alkaloids isolated from the stem of Erythroxylum caatingae Plowman. All test fractions, except the hexane fractions, showed antimicrobial activity on gram-positive bacteria and fungi. The acetate: methanol (95:5), acetate, chloroform and hexane fractions show the highest cytotoxicity activity against the NCI-H292, HEp-2 and K562 cell lines using MTT. The absence of hemolysis in the erythrocytes of mice was observed in these fractions and 6β-Benzoyloxy-3α-(3,4,5- trimethoxybenzoyloxy) tropane (catuabine B). Staining with Annexin V-FITC and JC-1 was used to verify the mechanism of action of the compounds of E. caatingae that showed cytotoxicity less than 30 μg/mL in leukemic cells. After 48 h of incubation, we observed that the acetate: methanol (95:5), acetate, and chloroform fractions, as well as the catuabine B, increased in the number of cells in early apoptosis, from 53.0 to 74.8%. An analysis of the potential of the mitochondrial membrane by incorporation of JC-1 showed that most cells during incubation of the acetate: methanol (95:5) and acetate fractions (63.85 and 59.2%) were stained, suggesting the involvement of an intrinsic pathway of apoptosis.
在该研究中,我们检测了从卡廷加古柯(Erythroxylum caatingae Plowman)茎中分离得到的甲醇提取物、各馏分、总生物碱馏分以及两种生物碱的抗肿瘤和抗菌活性。除己烷馏分外,所有测试馏分均对革兰氏阳性菌和真菌显示出抗菌活性。使用MTT法检测发现,乙酸乙酯:甲醇(95:5)、乙酸乙酯、氯仿和己烷馏分对NCI-H292、HEp-2和K562细胞系具有最高的细胞毒性活性。在这些馏分以及6β-苯甲酰氧基-3α-(3,4,5-三甲氧基苯甲酰氧基)托烷(卡图宾B)中,未观察到小鼠红细胞发生溶血现象。采用膜联蛋白V-FITC和JC-1染色来验证卡廷加古柯中细胞毒性小于30μg/mL的化合物在白血病细胞中的作用机制。孵育48小时后,我们观察到乙酸乙酯:甲醇(95:5)、乙酸乙酯和氯仿馏分以及卡图宾B使早期凋亡细胞数量增加,从53.0%增至74.8%。通过掺入JC-1对线粒体膜电位进行分析表明,在乙酸乙酯:甲醇(95:5)和乙酸乙酯馏分孵育期间,大多数细胞(分别为63.85%和59.2%)被染色,这表明凋亡的内在途径参与其中。