León-Sicairos Nidia, López-Soto Fernando, Reyes-López Magda, Godínez-Vargas Delfino, Ordaz-Pichardo Cynthia, de la Garza Mireya
Departamento de Biología Celular, Centro de Investigación y de Estudios Avanzados del I.P.N., Apdo. 14-740, México, D.F. 07000, México.
Clin Med Res. 2006 Jun;4(2):106-13. doi: 10.3121/cmr.4.2.106.
To identify amoebicidal components in human milk and the effect of iron on the amoebicidal activity.
Investigation in axenic cultures of Entamoeba histolytica trophozoites.
Amoebas were treated with 5%-20% of human, bovine and swine milk, with 10% of human milk fractions (i.e., casein, proteins except casein and fat) or with 1 mg/ml of human milk apo-lactoferrin, human secretory immunoglobulin type A (sIgA) and chicken egg-white lysozyme (i.e., purified proteins). Milk proteins were detected using immunoblot. Confocal microscopy was used to define the interaction of milk proteins (100 microM each) and amoebas. Experiments were done at least three times in triplicate, and mean and standard deviations were calculated.
Human and bovine milk were amoebicidal showing a concentration-dependent effect. The amoebicidal effect was increased in the absence of iron. Milk protein fractions, with the exception of casein, were the components responsible for the amoebicidal activity found. Apo-lactoferrin, sIgA and lysozyme were identified in the amoebicidal milk protein fraction. Apo-lactoferrin showed the major amoebicidal effect. These proteins, either alone or in combination, showed a killing effect on the trophozoites. They bound to the amoebic membrane causing cell rounding, lipid disruption and damage.
Milk proteins such as apo-lactoferrin, sIgA and lysozyme are able to kill Entamoeba histolytica trophozoites. This study confirms the importance of feeding breast milk to newborns.
鉴定人乳中的杀阿米巴成分以及铁对杀阿米巴活性的影响。
在溶组织内阿米巴滋养体的无菌培养物中进行研究。
用5%-20%的人乳、牛乳和猪乳,10%的人乳组分(即酪蛋白、除酪蛋白外的蛋白质和脂肪)或1mg/ml的人乳脱铁乳铁蛋白、人分泌型免疫球蛋白A(sIgA)和鸡卵清溶菌酶(即纯化蛋白)处理阿米巴。使用免疫印迹法检测乳蛋白。共聚焦显微镜用于确定乳蛋白(各100μM)与阿米巴之间的相互作用。实验至少重复三次,每次重复三次,并计算平均值和标准差。
人乳和牛乳具有杀阿米巴作用,呈浓度依赖性效应。在无铁的情况下,杀阿米巴作用增强。除酪蛋白外的乳蛋白组分是所发现的杀阿米巴活性的 responsible for 成分。在具有杀阿米巴活性的乳蛋白组分中鉴定出脱铁乳铁蛋白、sIgA和溶菌酶。脱铁乳铁蛋白显示出主要的杀阿米巴作用。这些蛋白质单独或组合使用时,对滋养体均有杀伤作用。它们与阿米巴细胞膜结合,导致细胞变圆、脂质破坏和损伤。
脱铁乳铁蛋白、sIgA和溶菌酶等乳蛋白能够杀死溶组织内阿米巴滋养体。本研究证实了给新生儿喂母乳的重要性。