菊花活性成分Parthenolide对人鼻咽癌CNE2细胞线粒体功能和半胱天冬酶活性的影响(英文)  

Influence of active ingredient of chrysanthemum Parthenolide on mitochondrial function of CNE2 cell of human nasopharyngeal carcinoma and caspase activity

作　　者：林忠宁 [1] 林育纯 [1] SHEN Han-ming  YANG Cheng-feng  Ong Choon-Nam  

机构地区：[1]中山大学公共卫生学院预防医学系,广东省 广州市 510089 [2]Department of Community,Occupational and Family Medicine,Faculty of Medicine,Faculty of Medcine,MD3,National University of Singapore,Singapore 117597

出　　处：《中国临床康复》2004年第5期979-981,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金课题(30070649);广东省自然科学基金课题(001369);美国中华医学基金会(CMB)课题;广东省医药管理局基金(103054)~~

摘　　要：背景:鼻咽癌是我国南方重点防治的恶性肿瘤之一,探讨天然草本菊花活性成分在NPC化学预防的作用,对建立现代康复体系具有重要意义。目的:研究菊花活性成分倍半萜烯内酯(SLs)对人鼻咽癌细胞中线粒体功能和半胱天冬酶活化途径的影响。设计:完全随机对照分组设计。地点和对象:由中山大学公共卫生学院预防医学系完成,对象为低分化CNE2细胞株。干预:对parthenolide(PN)进行剂量-反应和时间效应作用观察,采用噻唑蓝(MTT)颜色反应检测细胞线粒体功能,底物荧光光谱法测定细胞内caspase-9和-3活性,蛋白免疫印迹法检测线粒体内细胞色素C释放和caspase-3酶原降解片段;并应用特异性抑制剂进行细胞内cas-pase途径阻断实验。主要观察指标:细胞线粒体功能、细胞内caspase-9和caspase-3活性、线粒体细胞色素C的释放和caspase-3酶原的降解。结果:PN(1～100μmol/L)作用12h和24h后,MTT颜色反应抑制率随剂量显著增高,呈明显剂量依赖性(Pearson'sr=0.7322,0.7703,P<0.05),IC50分别为252.94μmol/L和49.63μmol/L;但caspase-9和-3活性未见升高,未见细胞色素C释放和caspase-3酶原降解片段形成。PN与caspase抑制剂联合作用后,caspase-9和-3活性明显低于对照和单独PN作用(t=9.146,8.280,27.325,27.450,P<0.05)。BACKGROUND:Parthenolide(PN) is the pricipal component of sesquiterpene lactones contained in some aromatic herbs.Nasopharyngeal carcinoma(NPC) occurs worldwide is one of highest incidence malignant tumor in south China.It is essential that using the PN as the therapy of health preserving of traditional Chinese medicine to develop the modern rehabilitation for NPC. OBJECTIVE:To study the effect of sesquiterpene lactones(SLs),the active ingredient of chrysanthemum,on the mitochondrial function of NPC cell and activating passage of caspase. DESIGN:Randomized controlled trial was conducted in this study. SETTING AND PARTICIPANTS:Completed by the Department of Preventive Medicine, College of Public Health, Sun Yat Sen University with object of poorly differentiated CNE2 cell strain. INTERVENTION:Parthenolide is given and the dose reaction and time effect are observed. The cellular mitochondrial function is detected with MTT color reaction, cellular caspase 9 and 3 activity were measured with substrate fluorescence spectrum, the release of mitochondrial cytochromic C(CytC) and cleavage segment of caspase 3 proenzyme were detected with protein immunity blotting; and specific inhibitor was applied for the blocking passage experiment of cellular caspase. MAIN OUTCOME MEASURE: cellular mitochondrial function, cellular caspase 9 and caspase 3 activity, release of mitochondrial CytC and cleavage of caspase 3 proenzyme. RESULTS:After acted by PN(1-100 μmol/L) for 12 hours and 24 hours, MTT color reaction inhibition rate grows obviously with the dose,indicating dose dependence (Pearson 's γ=0.7322,0.7703, P< 0.05),IC50 was 252.94 μmol/L and 49.63 μmol/L respectively; but without rising of caspase 9 and 3 activity, release of CytC and formation of caspase 3 proenzyme cleavage. Affected by PN and caspase inhibitor at same time,caspase 9 and 3 activity were apparently lower than that of control and single PN effect.(t=9.146, 8.280, 27.325, 27.450,P >0.05). CONCLUSION:PN may induce lowered function and deplet

关 键 词：菊花 活性成分 PARTHENOLIDE 人鼻咽癌 CNE2细胞 线粒体功能 半胱天冬酶活性 倍半萜烯内酯 

分 类 号：R273.96[医药卫生—中西医结合] R282.71[医药卫生—中医肿瘤科]