Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, 59 Mucai Street, Xiangfang District, Harbin 150030, People's Republic of China.
Biometals. 2010 Jun;23(3):555-61. doi: 10.1007/s10534-010-9324-2. Epub 2010 Mar 17.
Bovine lactoferricin (LfcinB) is a cationic peptide that selectively induces apoptosis in Jurkat cells. However less is known about the influence of this kind of apoptosis on the intra-cellular ceramide metabolism and the structure-function relationship between the loop structure of LfcinB and its action of inducing apoptosis in Jurkat cells. In the present study, the artificially synthesized LfcinB and LfcinB-derived peptide (Cys 19 residue in LfcinB was replaced by Ala) was added in Jurkat cells, the nucleolus shape was observed by fluorescent microscopy, the ceramide concentration in Jurkat cells was determined by reversed phase high performance liquid chromatography (RP-HPLC). The results of MTT assay showed that LfcinB inhibited proliferation of Jurkat cells, and the inhibition rate was approximately 18.90%. Moreover, the inhibition rate of LfcinB together with MAPP was upto approximately 59.89%. The RP-HPLC result showed that LfcinB improved the ceramide level in Jurkat cells. By using the DNA fragmentation assay and observing the nucleolus shape, the result displayed deficiency of the loop structure could cause LfcinB losing the biological activity of inducing apoptosis in Jurkat cells.
牛乳铁蛋白衍生抗菌肽(LfcinB)是一种阳离子肽,能选择性诱导 Jurkat 细胞凋亡。然而,对于这种凋亡对细胞内神经酰胺代谢的影响,以及 LfcinB 环结构与其诱导 Jurkat 细胞凋亡的作用之间的结构-功能关系,人们知之甚少。在本研究中,将人工合成的 LfcinB 和 LfcinB 衍生肽(LfcinB 中的 Cys19 残基被丙氨酸取代)添加到 Jurkat 细胞中,通过荧光显微镜观察核仁形态,用反相高效液相色谱法(RP-HPLC)测定 Jurkat 细胞中的神经酰胺浓度。MTT 检测结果显示,LfcinB 抑制 Jurkat 细胞增殖,抑制率约为 18.90%;同时添加 LfcinB 和 MAPP 时,抑制率约为 59.89%。RP-HPLC 结果显示,LfcinB 提高了 Jurkat 细胞中的神经酰胺水平。通过 DNA 片段化检测和观察核仁形态,结果显示环结构的缺失导致 LfcinB 丧失诱导 Jurkat 细胞凋亡的生物学活性。