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【目的】分析蛹拟青霉(Paecilomyces militaris)菌株RCEF0927发酵液中的抗中国仓鼠卵巢瘤(CHO)细胞活性及活性成分的具体组成。【方法】用刃天青(Resazurin)法测定样品对CHO细胞的抑制率;用高效液相色谱-高分辨质谱和活性测定联用的方法进行活性成分分析和鉴定。【结果】活性测定结果表明菌株RCEF0927的发酵液具有较强的抗CHO细胞活性;提取实验结果表明抗肿瘤活性物质能较好地被乙酸乙酯提取出来;液相色谱-质谱-活性测定分析表明提取物中活性组分的可能分子式分别为C10H13N5O3、C22H22O9、C23H24O10、C22H22O10、C15H10O4、C15H10O5、C36H70O11、C36H68O12、C38H74O11、C40H76O13和C44H82O14。【结论】根据细胞毒性、色谱特性、紫外光谱特征和质谱碎片,可初步推断发酵物中抗肿瘤活性成分分别为虫草素、4′-羟基异黄酮-7-O-(4″-O-甲基)-β-D-葡萄糖甙、6-甲氧基-4′-羟基异黄酮-7-O-(4″-O-甲基)-β-D-葡萄糖甙、5,4′,-二羟基异黄酮-7-O-(4″-O-甲基)β--D-葡萄糖甙、7,4′,-二羟基异黄酮、5,7,4′,-三羟基异黄酮、油酸酯聚氧乙烯醚、聚氧乙烯山梨醇酐单硬脂酸酯脱水化合物、聚氧乙烯山梨醇酐单油酸酯,以及含聚氧丙烯或聚氧丁烯结构单元的聚氧乙烯山梨醇酐脂肪酸酯衍生物等化合物。上述化合物对CHO细胞的毒性为首次发现。
【Objective】 The purpose of this study was to analyze the activity of Chinese hamster ovary ovary (CHO) cells in the fermentation broth of Paecilomyces militaris strain RCEF0927 and the specific composition of the active components. 【Method】 Resazurin method was used to determine the inhibition rate of CHO cells. The active ingredient was analyzed and identified by high performance liquid chromatography-high resolution mass spectrometry coupled with activity assay. 【Result】 The results of the activity assay showed that the fermentation broth of strain RCEF0927 had strong anti-CHO cell activity. The results of the extraction showed that the antitumor active substance could be extracted by ethyl acetate. The analysis by liquid chromatography-mass spectrometry The possible formulas of the active components in the extract are C10H13N5O3, C22H22O9, C23H24O10, C22H22O10, C15H10O4, C15H10O5, C36H70O11, C36H68O12, C38H74O11, C40H76O13 and C44H82O14, respectively. 【Conclusion】 According to the results of cytotoxicity, chromatographic characteristics, UV spectrum and mass spectrum, the antitumor activity of the fermented material can be inferred as cordycepin, 4’-hydroxyisoflavone-7-O- Β-D-glucoside, 6-methoxy-4’-hydroxyisoflavone-7-O- (4 “-O-methyl) Dihydroxyisoflavone-7-O- (4 ”-O-methyl) β-D-glucoside, 7,4 ’, - dihydroxyisoflavone, 5,7,4’ Oleate polyoxyethylene ether, polyoxyethylene sorbitan monostearate dehydrating compound, polyoxyethylene sorbitan monooleate, and polyoxyethylene sorbitan polyoxypropylene or polyoxybutylene containing structural units Alcohol anhydride fatty acid ester derivatives, etc. The above compound was found to be toxic to CHO cells for the first time.