罗非鱼头长链碱的分离纯化及抗肿瘤活性

Extraction, Purification and Anti-Tumor Activity in Vivo of Long Chain Bases from Tilapia Head

目的:探讨罗非鱼头长链碱(tilapia head long chain bases,TH-LCB)的提取纯化方法及其体外抗肿瘤活性。方法:以罗非鱼头为原料,采用有机溶剂提取法,提取纯化得到TH-LCB;采用四甲基偶氮唑盐法、细胞凋亡率和细胞周期的测定和蛋白免疫印迹等方法,探讨了TH-LCB对人白血病K562细胞的增殖抑制作用和凋亡诱导作用。结果:TH-LCB可显著抑制人K562细胞的增殖,TH-LCB作用24、48 h后的半数抑制浓度分别为42.207、39.494μg/m L;随着TH-LCB质量浓度的增加,细胞凋亡率逐渐增加,sub-G0/G1峰(凋亡峰)也随之升高,表明TH-LCB通过诱导细胞凋亡来抑制K562细胞增殖;Western blot结果表明,TH-LCB可提高细胞内Caspase-3的蛋白表达量,呈剂量依赖效应。结论:TH-LCB可通过诱导K562细胞凋亡抑制其增殖,这一过程可能与Caspase-3的激活有关。

Purpose:To investigate the extraction and purification of long chain bases from tilapia head(TH-LCB)and to assess their anti-tumor activity in vivo.Methods:The long chain bases were extracted into an organic solvent and purified.In addition,the anti-tumor activity of TH-LCB was detected by3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assay,Annexin V assay,cell cycle analysis and Western blot.Results:TH-LCB significantly reduced the viability of K562cells in a dose-dependent manner.After treatment with TH-LCB for24and48h,its half maximal inhibitory concentration were42.207and39.494μg/mL,respectively.TH-LCB increased the apoptotic rate,indicating that it reduced cell viability by inducing apoptosis in K562cells.Flow cytometric analysis confirmed that TH-LCB caused accumulation cells in the sub-G0/G1phase,being an indicative of apoptosis.Furthermore,TH-LCB also significantly increased the expression of caspase-3in a dose-dependent manner.Conclusion:TH-LCB can inhibit the proliferation of K562cells through inducing apoptosis,which may be correlated with the activation of caspase-3.