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为研究沙蚕提取物的体外抗肿瘤活性与机制,取海洋环节动物沙蚕(Nereis virens)冻干粉,经乙醇浸提、乙酸乙酯萃取,得到萃取物A1;经硅胶柱分段梯度洗脱分离得组分Bi,其中石油醚∶乙酸乙酯(体积比3∶7)洗脱的组分B4具有较好的抗肿瘤活性。采用制备型HPLC对B4进一步梯度洗脱分离得到Ci等5个组分。噻唑蓝(MTT)法测定表明,组分C4具有显著的体外抗A375细胞生长的活性,其IC50(48 h)值达到52.67μg/m L。AO/EB染色法观察细胞凋亡,显示药物处理48 h后细胞明显出现凋亡现象;通过流式细胞术及测定相对酪氨酸酶活力和黑色素含量,发现组分C4的抗肿瘤细胞A375机制以诱导细胞凋亡为主。
In order to study the anti-tumor activity and mechanism of Nereis extract in vitro, Nereis virens freeze-dried powder was extracted by ethanol and extracted with ethyl acetate to obtain extract A1. The residue was fractionated by silica gel column gradient The fraction B4, which is fractionated off Bi, has a good anti-tumor activity, with fraction B4 eluted with petroleum ether: ethyl acetate (3: 7 by volume). Preparative HPLC was used to separate and purify B4 to obtain 5 components such as Ci. The results of MTT assay showed that component C4 had significant anti-A375 cell growth activity in vitro, and its IC50 (48 h) reached 52.67μg / m L. Apoptosis was observed by AO / EB staining, which showed that the cells were obviously apoptotic after 48 h of treatment. The anti-tumor cell A375 mechanism of component C4 was found by flow cytometry and relative tyrosinase activity and melanin content To induce apoptosis mainly.