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目的:分析木蹄层孔菌(Fomes fomentarius)石油醚组分中甾醇和三萜类等成分及其主要抗肿瘤活性物质。方法:采用95%乙醇回流提取法及拌硅胶淋洗方法得到木蹄层孔菌的石油醚组分。用气相色谱-质谱联用技术对石油醚组分甾体和三萜类成分进行分析和鉴定并计算出其含量。用硅胶柱色谱法分离纯化含量最高的7,22-二烯麦角甾-3-酮。采用Alamar Blue法进行体外抗肿瘤活性检测。结果:从木蹄层孔菌石油醚组分中检测出37个成分,鉴定了其中的35个成分,包括10种甾体和三萜类成分。其主要成分为:7,22-二烯麦角甾-3-酮(37.67%)和5,24(28)-二烯麦角甾-3-醇(27.01%)。石油醚组分对细胞株NCI-H460的IC50为75μg.mL-1,7,22-二烯麦角甾-3-酮对细胞株NCI-H460的IC50为30μg.mL-1,抗肿瘤活性与单体浓度存在明显的量效关系。结论:木蹄层孔菌石油醚组分主要成分为7,22-二烯麦角甾-3-酮和5,24(28)-二烯麦角甾-3-醇,抗肿瘤活性物质最高的为7,22-二烯麦角甾-3-酮。研究结果为木蹄层孔菌的进一步开发提供了依据。
OBJECTIVE: To analyze the components of sterols and triterpenes in the petroleum ether fraction of Fomes fomentarius and its main anti-tumor active substances. Methods: Petroleum ether components were obtained from 95% ethanol reflux extraction and silica gel elution. Gas chromatography-mass spectrometry (GC-MS) was used to analyze and identify the components of the petroleum ether component steroids and triterpenoids. The highest content of 7,22-dienic ergosterol-3-one was isolated and purified by silica gel column chromatography. In vitro anti-tumor activity was detected by Alamar Blue method. Results: Thirty-seven components were detected from the petroleum ether fractions of Pseudomonas stomatalis and 35 of them were identified, including ten steroids and triterpenoids. Its main components are: 7,22-diene ergosterol-3-one (37.67%) and 5,24 (28) -dienylegasteride-3-ol (27.01%). The IC50 of the petroleum ether component to the cell line NCI-H460 was 75 μg.mL-1,7,22-dienicotetra-3-one had an IC50 of 30 μg.mL-1 against the cell line NCI-H460, the antitumor activity was Monomer concentration has a significant dose-effect relationship. CONCLUSION: The main constituents of the petroleum ether component of Pseudomonas stomatalis are 7,22-dienone ergosten-3-one and 5,24 (28) -dienyl ergostat-3-ol. The highest antitumor activity is 7,22-diene ergosterol-3-one. The results provided the basis for the further development of Fusarium.