柳叶亚菊中酚类化合物的抗氧化和抗肿瘤活性及凋亡机制  

A study involving antioxidizability and cytotoxicity of two kinds of phenol from Ajania Salicifolia and their mechanisms of apoptosis

作　　者：张伟[1,2] 巫洪儒[3] 梁乾坤 李云霞 卢研宇[2] 龙瑶[2] 祝英[3] 李红芳[1,2,4] 

机构地区：[1]兰州大学基础医学院生理学教研室 [2]甘肃省新药临床前研究重点实验室 [3]兰州大学应用有机化学国家重点实验室,甘肃兰州730000 [4]兰州大学基础医学院机能实验室

出　　处：《中国应用生理学杂志》2015年第5期422-426,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(21272103)

摘　　要：目的:从柳叶亚菊植物中提取的两种具有显著生物活性的天然苯酚类化合物4-羟基-3,5-二甲氧基-4-(2-丙酮基)环己烷-2,5-二烯-1-酮和6-甲氧基-5,7-二羟基香豆素(分别命名为Ⅰ号和Ⅱ号),分析测定其抗氧化活性和抗肿瘤活性及其促凋亡机制。方法:采用DPPH和ABTS自由基法以及MTT法分别测定两种化合物抗氧化和体外抗肿瘤活性,利用RT-PCR测定两种化合物对人血癌细胞株K562细胞核转录因子NF-κB P65 mRNA的表达以及Western blot测定其对核转录因子NF-κB P65、Fas、β-catenin和E-cadherin四种蛋白表达的影响。结果:1抗氧化实验分析得出Ⅰ号化合物具有强烈的抗氧化活性而Ⅱ号化合物无抗氧化活性;2抗肿瘤(MTT)实验分析数据可知Ⅰ号化合物无抗肿瘤活性而Ⅱ号化合物具有强烈的抗肿瘤活性;3利用实时定量q-PCR测得加入Ⅱ号化合物组与空白组相比,NF-κB P65 mRNA的表达明显上调(P<0.05);4通过Western blot实验得出Ⅱ号化合物组核转录因子NF-κB P65、Fas、β-catenin和E-cadherin四种蛋白的表达明显上调(P<0.05),并且蛋白表达量与药物浓度呈现明显剂量效应关系。结论:两种酚类化合物分别具有抗氧化和抗肿瘤活性,Ⅱ号化合物的抑癌作用机理可能一方面通过上调NF-κB P65、Fas表达,进入经典的Fas凋亡信号通路;另一方面,通过诱导β-catenin连环蛋白以及E-cadherin钙粘蛋白的表达上调,二者参与细胞间的粘连,对防止癌细胞的扩散和肿瘤的转移具有重大意义。Objective： To extract two kinds of phenols 4- hydroxy-3,5-dimethoxy-4-（2-oxopmpyl） cyclohexa-2, 5-dien-l-one and 6- methoxy-5,7-dihydroxy eoumarin （named as I and Ⅱ compounds respectively） from Ajania salicifolia and to investigate their antioxidation and cytotoxicity to tumors and explore their pro-apoptosis mechanism. Methods： The antioxidant activities of two compounds were assessed by ABTS and DPPH radical-scavenging assays. Two compounds were evaluated for their eytotoxicity against human chronic myelogenous leukemia （K562） cells using the MTT assay. The expression of NF-aB P65 mRNA in K562 apoptotic cells was measured by reverse transcription-poly- merase chain reaction （RT-PCR）, real-time quantitative PCR. In addition, protein expression levels of the NF-xB P65, p-Akt, Fas, β-cateni- na and E-cadherin were also measured by Western blot. Results： （1） We found that compound I displayed significant inoxidizability, while compound Ⅱ had no obvious antioxidizability. （2） In cytotoxicity experiments, compound I didn＇t display cytotoxicity while compound Ⅱ displayed obvious cytotoxieity. （3） Compared with the blank group, the expression of NF-xB P65 mRNA in K562 cell after treatment with compound Ⅱ was obviously up-regulated. （4） Compared with the blank group, the expression levels of NF-xB P65, Fas, β-catenina and E-cad- herin were significantly increased in compound n treated groups and it appeared obvious dose-effect relationship between the expression of pro- tein and drug concentration. Conclusion： Two phenols have obvious antioxidizability and cytotoxicity respectively. On the one hand, the tu- mor-suppressing mechanism of compound Ⅱ maybe act by up-regulation the expression of NF-kB 1365 and Fas protein; thereby, affecting the classical Fas apoptosis signaling pathways. On the other hand, it can also up-regulate the expression of protein β-eatenin and E-eadherin, which participate in the adhesion between cells, and accordingly, playing an important rol

关 键 词：酚类化合物 抗氧化活性 抗肿瘤活性 核转录因子-κB P65 FAS受体 Β-连环蛋白 E-钙粘蛋白 

分 类 号：R730.53[医药卫生—肿瘤]