López-Saiz Carmen-María, Velázquez Carlos, Hernández Javier, Cinco-Moroyoqui Francisco-Javier, Plascencia-Jatomea Maribel, Robles-Sánchez Maribel, Machi-Lara Lorena, Burgos-Hernández Armando
Departamento de Investigación y Posgrado en Alimentos, Universidad de Sonora, Apartado Postal 1658, Hermosillo, Sonora 83000, Mexico.
Departamento de Ciencias Químico-Biológicas, Universidad de Sonora, Apartado Postal 1685, Hermosillo, Sonora 83000, Mexico.
Int J Mol Sci. 2014 Dec 17;15(12):23555-70. doi: 10.3390/ijms151223555.
Shrimp is one of the most popular seafood items worldwide, and has been reported as a source of chemopreventive compounds. In this study, shrimp lipids were separated by solvent partition and further fractionated by semi-preparative RP-HPLC and finally by open column chromatography in order to obtain isolated antiproliferative compounds. Antiproliferative activity was assessed by inhibition of M12.C3.F6 murine cell growth using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay. The methanolic fraction showed the highest antiproliferative activity; this fraction was separated into 15 different sub-fractions (M1-M15). Fractions M8, M9, M10, M12, and M13 were antiproliferative at 100 µg/mL and they were further tested at lower concentrations. Fractions M12 and M13 exerted the highest growth inhibition with an IC50 of 19.5 ± 8.6 and 34.9 ± 7.3 µg/mL, respectively. Fraction M12 was further fractionated in three sub-fractions M12a, M12b, and M12c. Fraction M12a was identified as di-ethyl-hexyl-phthalate, fraction M12b as a triglyceride substituted by at least two fatty acids (predominantly oleic acid accompanied with eicosapentaenoic acid) and fraction M12c as another triglyceride substituted with eicosapentaenoic acid and saturated fatty acids. Bioactive triglyceride contained in M12c exerted the highest antiproliferative activity with an IC50 of 11.33 ± 5.6 µg/mL. Biological activity in shrimp had been previously attributed to astaxanthin; this study demonstrated that polyunsaturated fatty acids are the main compounds responsible for antiproliferative activity.
虾是全球最受欢迎的海鲜之一，并且据报道是化学预防化合物的一个来源。在本研究中，虾脂质通过溶剂分配进行分离，然后通过半制备RP-HPLC进一步分级，最后通过开放柱色谱法进行分级，以获得分离的抗增殖化合物。使用MTT（3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐）法通过抑制M12.C3.F6小鼠细胞生长来评估抗增殖活性。甲醇提取物显示出最高的抗增殖活性；该提取物被分离成15个不同的亚级分（M1-M15）。M8、M9、M10、M12和M13级分在100μg/mL时具有抗增殖作用，并在较低浓度下进一步测试。M12和M13级分表现出最高的生长抑制作用，IC50分别为19.5±8.6和34.9±7.3μg/mL。M12级分进一步分离成三个亚级分M12a、M12b和M12c。M12a级分被鉴定为邻苯二甲酸二乙基己酯，M12b级分为至少被两种脂肪酸取代的甘油三酯（主要是油酸并伴有二十碳五烯酸），M12c级分为另一种被二十碳五烯酸和饱和脂肪酸取代的甘油三酯。M12c中含有的生物活性甘油三酯表现出最高的抗增殖活性，IC50为11.33±5.6μg/mL。虾中的生物活性先前归因于虾青素；本研究表明多不饱和脂肪酸是抗增殖活性的主要化合物。
