牡蛎共生菌Chaetomium globosum ML4发酵液化学成分及其体外抗肿瘤活性研究.docVIP

牡蛎共生菌Chaetomium globosum ML4发酵液化学成分及其体外抗肿瘤活性研究 　　[摘要]采用硅胶柱色谱、Sephadex LH20凝胶柱色谱、制备薄层色谱和高效液相色谱等方法对牡蛎共生菌Chaetomium globosum ML4发酵液的化学成分进行分离纯化，获得5个化合物。利用高分辨质谱、核磁共振谱及文献比对的方法，将上述化合物分别鉴定为chaetoglobosin V（1），chaetoglobosin Vb（2），tyrosol（3），5methyluracil（4），uracil（5）。采用MTT法测定化合物对人肝癌细胞株SMMC7721的体外细胞毒活性，结果发现，化合物1对SMMC7721细胞具有明显的增殖抑制活性，其IC50为605 mg?L-1，阳性对照药顺铂的IC50为1996 mg?L-1。进一步研究发现，化合物1可引起SMMC7721细胞G2期细胞周期阻滞，并能诱导SMMC7721细胞凋亡；SMMC7721细胞中Bcl2/Bax的比值下降，Caspases3，8，9的表达增加，Akt蛋白的表达和磷酸化水平下降。上述结果表明，化合物1对SMMC7721细胞的体外抗肿瘤活性与诱导细胞周期G2期阻滞以及细胞凋亡相关；其诱导凋亡作用同时涉及线粒体途径和死亡受体途径，且与PI3K/Akt信号通路活性下降相关。 　　[关键词]牡蛎； 共生菌； Chaetomium globosum； 化学成分； 抗肿瘤活性 　　[Abstract]Isolation and purification of chemical constituents of liquid culture of symbiotic Chaetomium globosum ML4 of oyster was performed through silica gel column chromatography， gel filtration over Sephadex LH20， preparative TLC and HPLC Five compounds were obtained and their structures were determined as chaetoglobosin V（1）， chaetoglobosin Vb（2）， tyrosol（3）， 5methyluracil（4）and uracil（5）， respectively， based on HRMS and NMR data and comparison with literatures In vitro cytotoxicity of compounds against human hepatocellular carcinoma cell line SMMC7721 were measured byMTT method， and results showed that compound 1 could obviously inhibit the proliferation of SMMC7721 cells with an IC50 value of 605 mg?L-1， while the IC50 value of positive control cisplatin was 1996 mg?L-1 Further studies discovered that compound 1 could lead to G2 phase arrest in SMMC7721 cells and induce SMMC7721 cells apoptosis The ratio of Bcl2/Bax in SMMC7721 cells was decreased The expression of protein Caspases3，8，9 was improved and the expression and phosphorylation level of Akt were reduced Aforementioned results revealed that in vitro antitumor activity of compound 1 against SMMC7721 cells were related to G2 phase cell cycle arrest and inducedapoptosis The inducedapoptosis was involved in both the mitochondrial pathway and the death receptor pathway and connected