瑞香素体外抗肿瘤及毒理学实验研究
摘要
目的:初步探讨金边瑞香活性成分瑞香素的体外抗肿瘤活性,并对其进行安全性评价,为金边瑞香活性成分的药用开发提供重要依据。
方法: 1.采用MTT法进行瑞香素体外抑瘤试验,检测不同浓度瑞香素对人肝癌SMMC-7721细胞和小鼠S180细胞增殖的影响。2.通过急性毒性试验、鼠伤寒沙门氏菌/哺乳动物微粒体酶试验(Ames试验)、骨髓细胞微核试验对瑞香素进行安全性评价。
结果:1.瑞香素浓度(μg/ml)为:1.95、3.90、7.81、15.62、31.25、62.5、125、250时,作用于人肝癌SMMC-7721细胞,瑞香素抑制率(%)分别为:51.48±1.65、60.32±2.49、68.97±2.51、74.23±2.56、78.51±0.61、79.35±1.22、83.6±1.47、87.1±2.35;作用于小鼠S180细胞时,瑞香素抑制率(%)分别为7.52±3.70、14.96±4.90、38.17±12.22、40.64±12.52、52.01±2.67、69.10±4.31、76.28±10.62、86.54±4.24。瑞香素对SMMC-7721和S180细胞增殖抑制作用明显,并呈现良好的剂量依赖性。2.以超过人体有效剂量175倍剂量100mg/kg.BW,一次性灌胃小鼠,连续观察7天未见中毒体征,体重增长正常。3.瑞香素Ames试验各剂量组回变菌落数均未超过阴性对照组菌落数2倍,亦无剂量反应关系。瑞香素Ames试验为阴性。4.瑞香素骨髓微核实验各剂量组微核率与阴性对照组比较无显著性差异(P>0.05),而环磷酰胺阳性对照组与阴性对照组及各样品剂量比较统计学有显著性差异(P<0.05)。表明瑞香素对小鼠骨髓细胞无抑制作用和遗传毒性,实验结果为阴性。
结论:1.瑞香素浓度(μg/ml)为:1.95、3.90、7.81、15.62、31.25、62.5、125、250范围内对SMMC-7721和S180细胞生长均有不同程度的抑制作用,并且呈现良好的剂量依赖性。
2.瑞香素在本试验剂量范围内未见明显的毒副作用,无致突变作用,安全性较高。
Objective: To evaluate the toxicity of daphnetin, to study its antitumor activity furtherly, and to provide the theory foundation for the development of daphnetin as medicine.
Method: 1. Developing the MTT assay in the vitro of daphnetin, and studying the effect of daphnetin with different concentration to the SMMC-7721 and S180 cells when were proliferating. 2.Evaluating the toxicity of daphnetin by acute toxicity tests, Ames tests, and micronucleus test.
Results: 1. When the daphnetin worked on SMMC-772 cells with the concentration of (μg/ml) 1.95、3.90、7.81、15.62、31.25、62.5、125、250, the daphnetin inhibitory rates were51.48±1.65、60.32±2.49、68.97±2.51、74.23±2.56、78.51±0.61、79.35±1.22、83.6±1.47、87.1±2.35; when the daphnetin worked on S180 cells, the daphnetin inhibitory rates were7.52±3.70、14.96±4.90、38.17±12.22、40.64±12.52、52.01±2.67、69.10±4.31、76.28±10.62、86.54±4.24 respectively.
2. Mice were treated with the daphnetin with 100/kg.bw over 175 times to human’dose which were lasting 7 days without detecting any toxic signs and change of weights.
3. The reverse mutation of daphnetin with different concentration were less than twice times to the control group without dose-dependent action and the Ames array were negative.
4. There was no significance between the daphnetin groups and the control groups(p>0.05), thus significance observed between the CY group and negative group(p<0.05), which indicated that daphnetin had no inhibitory and inherent toxicity activity on morrow cells.
Conclusion: 1. The daphnetin had inhibitory activity to SMMC-7721和S180 cells in some range with dose-dependent effect. 2. The daphnetin had no significantly toxicity activity without mutagenesis.
方法: 1.采用MTT法进行瑞香素体外抑瘤试验,检测不同浓度瑞香素对人肝癌SMMC-7721细胞和小鼠S180细胞增殖的影响。2.通过急性毒性试验、鼠伤寒沙门氏菌/哺乳动物微粒体酶试验(Ames试验)、骨髓细胞微核试验对瑞香素进行安全性评价。
结果:1.瑞香素浓度(μg/ml)为:1.95、3.90、7.81、15.62、31.25、62.5、125、250时,作用于人肝癌SMMC-7721细胞,瑞香素抑制率(%)分别为:51.48±1.65、60.32±2.49、68.97±2.51、74.23±2.56、78.51±0.61、79.35±1.22、83.6±1.47、87.1±2.35;作用于小鼠S180细胞时,瑞香素抑制率(%)分别为7.52±3.70、14.96±4.90、38.17±12.22、40.64±12.52、52.01±2.67、69.10±4.31、76.28±10.62、86.54±4.24。瑞香素对SMMC-7721和S180细胞增殖抑制作用明显,并呈现良好的剂量依赖性。2.以超过人体有效剂量175倍剂量100mg/kg.BW,一次性灌胃小鼠,连续观察7天未见中毒体征,体重增长正常。3.瑞香素Ames试验各剂量组回变菌落数均未超过阴性对照组菌落数2倍,亦无剂量反应关系。瑞香素Ames试验为阴性。4.瑞香素骨髓微核实验各剂量组微核率与阴性对照组比较无显著性差异(P>0.05),而环磷酰胺阳性对照组与阴性对照组及各样品剂量比较统计学有显著性差异(P<0.05)。表明瑞香素对小鼠骨髓细胞无抑制作用和遗传毒性,实验结果为阴性。
结论:1.瑞香素浓度(μg/ml)为:1.95、3.90、7.81、15.62、31.25、62.5、125、250范围内对SMMC-7721和S180细胞生长均有不同程度的抑制作用,并且呈现良好的剂量依赖性。
2.瑞香素在本试验剂量范围内未见明显的毒副作用,无致突变作用,安全性较高。
Objective: To evaluate the toxicity of daphnetin, to study its antitumor activity furtherly, and to provide the theory foundation for the development of daphnetin as medicine.
Method: 1. Developing the MTT assay in the vitro of daphnetin, and studying the effect of daphnetin with different concentration to the SMMC-7721 and S180 cells when were proliferating. 2.Evaluating the toxicity of daphnetin by acute toxicity tests, Ames tests, and micronucleus test.
Results: 1. When the daphnetin worked on SMMC-772 cells with the concentration of (μg/ml) 1.95、3.90、7.81、15.62、31.25、62.5、125、250, the daphnetin inhibitory rates were51.48±1.65、60.32±2.49、68.97±2.51、74.23±2.56、78.51±0.61、79.35±1.22、83.6±1.47、87.1±2.35; when the daphnetin worked on S180 cells, the daphnetin inhibitory rates were7.52±3.70、14.96±4.90、38.17±12.22、40.64±12.52、52.01±2.67、69.10±4.31、76.28±10.62、86.54±4.24 respectively.
2. Mice were treated with the daphnetin with 100/kg.bw over 175 times to human’dose which were lasting 7 days without detecting any toxic signs and change of weights.
3. The reverse mutation of daphnetin with different concentration were less than twice times to the control group without dose-dependent action and the Ames array were negative.
4. There was no significance between the daphnetin groups and the control groups(p>0.05), thus significance observed between the CY group and negative group(p<0.05), which indicated that daphnetin had no inhibitory and inherent toxicity activity on morrow cells.
Conclusion: 1. The daphnetin had inhibitory activity to SMMC-7721和S180 cells in some range with dose-dependent effect. 2. The daphnetin had no significantly toxicity activity without mutagenesis.
引文
[1] Shahid A. Khan1, Simon D.Taylor-Robinson1,Mireille B.Toledano , et al. Changing international trends in mortality rates for liver,biliary and pancreatic tumours[J]. Journal of Hepatology, 2002,37:806-813.
[2] Li Lian-di,Ru Feng-zhu, Zhang Si-wei,et al.Analysis and predicative analysis of Malignant tumor distribution and incidence in China in recent twenties[J].Chinese Journal of oncology, 1997;19(1):3-9.
[3] Dong Zhi-wei,Qiao You-lin,Li Lian-di,et al.Report of Chinese Cancer Control Stratey[J]. Bulletin of Chinese Cancer,2002,11(5):250-260.
[4] Cragg GM, NewmanDJ,Weiss RB.Coral reefs,forests,and the thermal vents:the worldwide exploration of nature for novel antitumor agents[J].SeminOneol,1997,24(2):156-163.
[5] Jimeno JM,Faircloth G,Gameron L,et al.Progress in the acquisition of new marine-derived anticancer compounds:development of ecteinascidin-743(ET-743) [J].Drugs Future,1996, 21:1155-1165Future,1996,21:1155-1165
[6] Row ley JD,Le Beau M M.Treatment related acutemyelid leukemia:new insight in Leukemogensis[J]. Mutat Res,1991,252:167
[7] KantarjianHM, Keating MJ.TheraPy-related leukemia and myelodysplastic syndrome[J]. Seymin Oncol,1987,14(4):435-443
[8] Povirk LF,Austin MJ.Genotoxicity of bleomycin[J].Mutat Res,1991(273):127-143
[9] Morita S,OhashiY,Hirakawa K,etal.PhaseⅠ/Ⅱstudy of combination therapy of oral fluorinated pyrimidine compound s-1with low- dose cisplatin in patients with unresectable and/or recurrent advanced gastric cancer:a novel study design to evaluate Safety and efficacy[J].Jpn J Clin Oncol,2004,34(3):159
[10]叶和杨,曾靖,杨敬格,等.金边瑞香的抗炎症作用[J].赣南医学院学报,2002,22(l):7.
[11]叶和杨,马廉兰,张瑞其,等.金边瑞香的抑菌实验[J].赣南医学院学报,2002,22(5):470-471.
[12]叶和杨,熊小琴,王中平等.金边瑞香提取液镇痛作用的实验研究[J].赣南医学院学报, 2004,24(5):505-507
[13]陈水亲,徐静,黄彬红,等.植物药金边瑞香对小鼠Peyer’s结的影响[J].中国临床康复, 2006, 10(3):85-89
[14]叶和杨,周俐,曾靖,等.金边瑞香对小鼠SOD活性及MDA含量的影响[J].赣南医学院学报, 2003, 23(3):246.
[15]徐静,傅颖媛.金边瑞香对小鼠免疫细胞的影响[J].广东医学,2005,26(9):1207-1208.
[16]徐静,傅颖媛.金边瑞香对小鼠免疫功能影响的量效关系[J].中国临床康复,2005,9 (43):103.
[17]黄彬红,傅颖媛,徐静,等.金边瑞香对荷S180瘤小鼠免疫功能的影响[J].时珍国医国药,2008,19(10):2376-2377
[18]舒奇,傅颖媛,刘婷,余燕影.金边瑞香活性成分瑞香黄烷-Ⅰ,瑞香素对SMMC-7721和MCF-7细胞作用的研究[J].时珍国药医药,2009,20(9):2149-2150.
[19] Zhang W. Studies on the active constituents of three medicinal plants of the Genus Daphne.[D], 2006
[20] Zhang W. Shen Y, Liu R, et al. Coumarins from the bark of Daphne marginata[J].Chemistry of Natural Compounds,2007,3(43):317-318
[21] W Zhang, W D Zhang, R Liu, et al. Two new chemical constituents from Daphne odora Thunb. var. marginata[J].Nat Prod Res,2006,20(14):1290
[22]陈瑞杰,金边瑞香化学成分研究[D] 2007,4:80-86
[23] Cottiglia F, Loy G, Garau D, Floris C,et al. Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L[J].Phytomedicine,2001,8 (4):302-305
[24] Qin Gao, Jinjun Shan, Liuqing Di,et al.Therapeutic effects of daphnetin on adjuvant-induced arthritic rats. [J]Journal of Ethnopharmacology.2008,(120) 259–263
[25] Zheng WF,Cao XW,Gu Q,et al,Antitumor acivity of daphnodorins from Daphen genkwa roots.[J]International Immunophar macology,2007,7:128-134
[26]毕跃峰,郑晓珂.卷柏属植物化学成分与药理活性[J].国外医药植物药分册,2002,17(3):97
[27] Misty Prince, Yan Li, Asper Childers, et al.Comparison of citrus coumarins on carcinogen-detoxifying enzymes in Nrf knockout mice[J].Toxicology Letters,2009,185:180– 186.
[28]司徒镇强,吴军正主编.细胞培养[M].世界图书出版公司,2007.
[29]路向前,张磊,蒋敬庭等.乳腺癌浸润淋巴细胞的细胞毒活性研究[J].江西医学检验,2002,12(6):361.
[30] Chiba T, Yokosuka O, Fukai K, et al. Cell growt h inhibition and gene expression induced by the histone deacetylase inhibitor, trichostatin A, on human hepatoma cell. [J]Oncology, 2004, 66:481-491.
[31] GB15193.3-2003,食品安全性毒理学评价程序和方法[S].
[32] GB15193.4-2003,食品安全性毒理学评价程序和方法[S].
[33] GB15193.5-2003,食品安全性毒理学评价程序和方法[S].
[34] Mosmann T.Rapid colorimetric assay for cellular growth and survival:Application to proliferation and cytotoxicity assay[J]. J Immunol Methods,1983,65:55-63
[35]新药(西药)临床前研究指导原则汇编.中华人民共和国卫生部药政局,1993,137-139.
[36] SehmidC,RotenbergJS.Neurodevelopmentaltoxieology[J].NeurolClin,2005,23(2):321-336.
[37] Holstege CP,DobmeierS. Cardiovascular challenges in toxieology[J].Emerg Med Clin NorthAm,2005,23(4):1195一1217
[38]陈奇主编.中药药理研究方法学[M].第二版.北京,人民卫生出版社,2006.131
[39] DostFN.Aeute toxieology of components of vegetation smoke[J]. Rev Environ Contam Tox1col,1991(119):l-46
[40] Bruekner,J.V.,KyleG.M.,Luthra,R.,etal.Aeute,short-term,and subchronic oraltoxieity1 of 1,1,l–trichloroethane in rats[J].Toxieol.Sei,2001(60):363-372
[41]周立国.药物毒理学[M].武汉:湖北科学技术出版社,2001.88-120
[42] Ames BN,Mccann J,Yamasaki E.Methods for detecting careinogens and mutagens with the Salmonella/mammalian-microsome mutagenieity test[J].MutatReS,1975(31):347一364
[43]王治乔,袁伯俊.新药临床安全性评价与实践[M].军事医学科学出版社,1997
[44] Rota PA,Obserste MS Monroe ss,etal.Characterization of a novel eoronavirus associated with severe acute respiratory syndrome[J]. Scienee,2003,300(5024):1394-1399
[45] Dayan J,Deguingand S,Truzman C,Study of the mutagenic activity of 6 hepatotoxic Pharmaeeutical drugs in the Salmonella typhimmurium microsome test,and the HGPRT and Na/K ATPase system incultured mammalian cells[J].MutatRes,1985(157):1-12
[46] Maron DM, Ames BN.Revised methods for the Sa1mon11a mutagenicity test[J].Mutation Res,1983(113):173
[47] Preston RJ,Au W,Bender, MA; et al. Mammalian in vivo and in vitro cytogenetic assays: a report of the U.S. EPA's Gene-Tox program[J].Mutat Res,1981(87):143-188
[48] Cottiglia F,Loy G,Garau D,et al.Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L[J].phytomedicine,2001,8(4):302-305.
[49]叶和杨,马廉兰,张瑞其,等.金边瑞香的抑菌试验[J]赣南医学院学报,2002,22(5):470-471
[50] Mohanmed AH.Cytological effect of Sudium fluoride on mice[J].Fluoride, 1982,15(3): 110-112
[51] HongSK,Kitano H,Satoh H,Nagato Y.How is embryo size genetically regulated in rice[J].Development,1996,122(7):2051-2058
[52]吕晓华.体外微核试验方法研究进展[J].癌变、畸变、突变,1999,11(2):109-112
[53] Mobamly MG.Cytogenetic testing of mutagenic Potential of the blue-green bacteriam,pleetonema boryanum in experimentally treated miee[J].Cytologia,1993,58(2):161-169
[1] Curtis D Klaassen. Casarett & Doull’s Toxicology :the Basic Science of Poisons[M] . McGraw2Hill ,2001. 1-104.
[2]沈芸,陆荣柱,陈自强.应用生态化标志物评价神经毒物的效应[J] .职业卫生与应急救援,2001 ,19 (3) :124 - 125.
[3] Letz R. Continuing Challenges for Computer2based Neuropsycho2 logical Tests[J ] . Neurotoxiclogy ,2003 ,24 (4 - 5) :479 - 489.
[4] Liu B ,Gao HM ,Hong J S. Parkinson s disease and exposure to infectious agents and pesticides and the occurrence of brain injuries :role of veuroinflammation[J] . Environ Health Perspect ,2003 ,111 (8) :1065 - 1073.
[5] Rohlman DS ,Gimenes LS , Eckerman DA ,et al. Development of the Behavioral Assessment and Research System (BARS) to Detect and Characterize neurotoxicity in Humans [J ] . Neurtoxicology ,2003 ,24 (4 - 5) :523 - 531.
[6] Bellinger DC. Perspectives on incorporating human neurobehavioral end points in risk assessments [ J ] . Risk Anal ,2002 ,22 ( 3) : 487 - 498.
[7]张晓玲.浅谈中药毒副作用[J].河南中药,2002,22 (2):10—71.
[8]袁何永.由马兜铃酸致肾病谈中药对肾的毒副作用[J].广东药学,2003,13(3):5961.
[9]陈慧袱.雷公藤的毒副作用及防治[J].浙江中医学院学报,1998,22(1):46.
[10]王新红.长春新碱引起神经毒副反应l2例临床分析[J].河南神经疾病药理学杂志,2002,5(4):68.
[11]钱伟平.蟾酥对实验动物循环系统毒副作用的研究[J].浙江中医学院学报,2001,25(6):46—47.
[12]高芳.中药的毒副作用及安全用药[J].中兽医医药-47-杂志,2000,(2):43—44.
[13]胡志祥,肖金莲主编.含毒中药毒理分析[J].时珍国医国药,1999,10(2):96.
[14]陈敬炳.天然药物中的毒蛋白、多肽及氨基酸的研究概况[J].中成药,2001,23(10):751—753.
[15]张东玲,张秀英.中药毒理学的研究进展[J].中兽医医药杂志,2005,1:51
[16]孙圆嫒,高月.中药毒性的研究概况[J].航空航天医药,2003,14(4):253
[17]马利华.浅析有毒中药及毒副作用[J].时珍国医国药,2004,15(6):377.
[18]马晓彤,张翠萍,张俊刚.松花粉片的急性毒性及致突变性研究[J].医学研究与教育,2009,26(3):16-17.
[19]舒奇,傅颖媛,刘婷,余燕影.金边瑞香活性成分瑞香黄烷-Ⅰ,瑞香素对SMMC-7721和MCF-7细胞作用的研究[J].时珍国药医药,2009,20(9):2149-2150.
[20]周爱香.复方马钱子片的镇痛作用和毒理试验[J].中国实验方剂学杂志,1998,4(4):59—60.
[21] Humphries ML , Olden K.Aspraginelineked oligosaccha—fide and tumor metastatis. Pharmac[J].Ther,1989,44:85.
[22] Denms Jw,Koch K,Youselfis,eta1.Growth hibition of humaelanoma tumor xenograftesi nathymic nude mice by swainso Cancer [J].Res,1990,50:1867～1872.
[23]刘锦霞.苦豆子、铁棒锤生物碱对菜青虫的毒力测定[J].甘肃科学学报,2003,15(2):38～39.
[24]鲁小青.蓖麻毒蛋白糖脂质体对肝癌细胞的杀伤作用及其理化特性[J].上海铁道大学学报,1998,19(9):24～25.
[25]高宝岩.蓖麻毒蛋白的提取及分析[J].光谱实验室,2001,l8(4):126～127.
[26]施大钊.毒芹生物碱的提取及对害鼠的毒性实验[J].植保技术与推广,2002,22(6):27～29.
[27]马丽.毒性中药与以毒攻毒[J].现代中西医结合杂志,2001,1o(23):2324～2325.
[28]刘红云.蜂毒的研究进展及其临床应用[J].中药材,2003,26(6):457～458.
[29]张坤娟.瑞香狼毒甲醇提取物抗瘤免疫机理初探[J].中国免疫学杂志.2003,19:544.
[30]吴正杰.“十八反”“十九畏”新探—兼论中药配伍-禁忌[J].中兽医医药杂志,2001,(3):21.
[31]吴慧英.谈十九畏用于临床[J].江西中医药,1997,28(2):44.
[32] Purchase IF,Botham PA,Bmner LH,eta1. workshopoverview:scientificandregula tory challenges for the reduction,refine ment,and replacement of animals in toxic ity testing[J].Toxicol Sci,1998,43(2): 86～101.
[33] Cheek B J,Stel A B,Torres M P,eta1.Chemiluminescence detection for hybridization assays on the flow—thru chip,a three dimensional microchannel biochip[J].Anal Chem,2001,15(24):5777-5783.
[34] Pennie W D.Use of cDNA microarray to probe and understand The toxicological consequences of altered gene expression [J]. Toxicological Le tters,2000,112—113:473-474.
[35] Nuwaysir E F,Bittner M,Trent J,et a1.Microarrays and toxicology:the advent of toxicogenomics[J].Mol Carcinog,1999, 24(3):153—159.
[36] Kiela P R Midura A J,Kuscuoglu N,et a1.Efects of Boswellia serrata in mouse models of chemically induced colitis[J].Am J Physiol Gastrointest Liver Physiol,2005,288(4):G798一G808.
[37]曹煜冲药复方物质基础和药效、毒性相关性研究[J]世界科学技术-中药现代化·基础研究,1999,1(2):36.
[2] Li Lian-di,Ru Feng-zhu, Zhang Si-wei,et al.Analysis and predicative analysis of Malignant tumor distribution and incidence in China in recent twenties[J].Chinese Journal of oncology, 1997;19(1):3-9.
[3] Dong Zhi-wei,Qiao You-lin,Li Lian-di,et al.Report of Chinese Cancer Control Stratey[J]. Bulletin of Chinese Cancer,2002,11(5):250-260.
[4] Cragg GM, NewmanDJ,Weiss RB.Coral reefs,forests,and the thermal vents:the worldwide exploration of nature for novel antitumor agents[J].SeminOneol,1997,24(2):156-163.
[5] Jimeno JM,Faircloth G,Gameron L,et al.Progress in the acquisition of new marine-derived anticancer compounds:development of ecteinascidin-743(ET-743) [J].Drugs Future,1996, 21:1155-1165Future,1996,21:1155-1165
[6] Row ley JD,Le Beau M M.Treatment related acutemyelid leukemia:new insight in Leukemogensis[J]. Mutat Res,1991,252:167
[7] KantarjianHM, Keating MJ.TheraPy-related leukemia and myelodysplastic syndrome[J]. Seymin Oncol,1987,14(4):435-443
[8] Povirk LF,Austin MJ.Genotoxicity of bleomycin[J].Mutat Res,1991(273):127-143
[9] Morita S,OhashiY,Hirakawa K,etal.PhaseⅠ/Ⅱstudy of combination therapy of oral fluorinated pyrimidine compound s-1with low- dose cisplatin in patients with unresectable and/or recurrent advanced gastric cancer:a novel study design to evaluate Safety and efficacy[J].Jpn J Clin Oncol,2004,34(3):159
[10]叶和杨,曾靖,杨敬格,等.金边瑞香的抗炎症作用[J].赣南医学院学报,2002,22(l):7.
[11]叶和杨,马廉兰,张瑞其,等.金边瑞香的抑菌实验[J].赣南医学院学报,2002,22(5):470-471.
[12]叶和杨,熊小琴,王中平等.金边瑞香提取液镇痛作用的实验研究[J].赣南医学院学报, 2004,24(5):505-507
[13]陈水亲,徐静,黄彬红,等.植物药金边瑞香对小鼠Peyer’s结的影响[J].中国临床康复, 2006, 10(3):85-89
[14]叶和杨,周俐,曾靖,等.金边瑞香对小鼠SOD活性及MDA含量的影响[J].赣南医学院学报, 2003, 23(3):246.
[15]徐静,傅颖媛.金边瑞香对小鼠免疫细胞的影响[J].广东医学,2005,26(9):1207-1208.
[16]徐静,傅颖媛.金边瑞香对小鼠免疫功能影响的量效关系[J].中国临床康复,2005,9 (43):103.
[17]黄彬红,傅颖媛,徐静,等.金边瑞香对荷S180瘤小鼠免疫功能的影响[J].时珍国医国药,2008,19(10):2376-2377
[18]舒奇,傅颖媛,刘婷,余燕影.金边瑞香活性成分瑞香黄烷-Ⅰ,瑞香素对SMMC-7721和MCF-7细胞作用的研究[J].时珍国药医药,2009,20(9):2149-2150.
[19] Zhang W. Studies on the active constituents of three medicinal plants of the Genus Daphne.[D], 2006
[20] Zhang W. Shen Y, Liu R, et al. Coumarins from the bark of Daphne marginata[J].Chemistry of Natural Compounds,2007,3(43):317-318
[21] W Zhang, W D Zhang, R Liu, et al. Two new chemical constituents from Daphne odora Thunb. var. marginata[J].Nat Prod Res,2006,20(14):1290
[22]陈瑞杰,金边瑞香化学成分研究[D] 2007,4:80-86
[23] Cottiglia F, Loy G, Garau D, Floris C,et al. Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L[J].Phytomedicine,2001,8 (4):302-305
[24] Qin Gao, Jinjun Shan, Liuqing Di,et al.Therapeutic effects of daphnetin on adjuvant-induced arthritic rats. [J]Journal of Ethnopharmacology.2008,(120) 259–263
[25] Zheng WF,Cao XW,Gu Q,et al,Antitumor acivity of daphnodorins from Daphen genkwa roots.[J]International Immunophar macology,2007,7:128-134
[26]毕跃峰,郑晓珂.卷柏属植物化学成分与药理活性[J].国外医药植物药分册,2002,17(3):97
[27] Misty Prince, Yan Li, Asper Childers, et al.Comparison of citrus coumarins on carcinogen-detoxifying enzymes in Nrf knockout mice[J].Toxicology Letters,2009,185:180– 186.
[28]司徒镇强,吴军正主编.细胞培养[M].世界图书出版公司,2007.
[29]路向前,张磊,蒋敬庭等.乳腺癌浸润淋巴细胞的细胞毒活性研究[J].江西医学检验,2002,12(6):361.
[30] Chiba T, Yokosuka O, Fukai K, et al. Cell growt h inhibition and gene expression induced by the histone deacetylase inhibitor, trichostatin A, on human hepatoma cell. [J]Oncology, 2004, 66:481-491.
[31] GB15193.3-2003,食品安全性毒理学评价程序和方法[S].
[32] GB15193.4-2003,食品安全性毒理学评价程序和方法[S].
[33] GB15193.5-2003,食品安全性毒理学评价程序和方法[S].
[34] Mosmann T.Rapid colorimetric assay for cellular growth and survival:Application to proliferation and cytotoxicity assay[J]. J Immunol Methods,1983,65:55-63
[35]新药(西药)临床前研究指导原则汇编.中华人民共和国卫生部药政局,1993,137-139.
[36] SehmidC,RotenbergJS.Neurodevelopmentaltoxieology[J].NeurolClin,2005,23(2):321-336.
[37] Holstege CP,DobmeierS. Cardiovascular challenges in toxieology[J].Emerg Med Clin NorthAm,2005,23(4):1195一1217
[38]陈奇主编.中药药理研究方法学[M].第二版.北京,人民卫生出版社,2006.131
[39] DostFN.Aeute toxieology of components of vegetation smoke[J]. Rev Environ Contam Tox1col,1991(119):l-46
[40] Bruekner,J.V.,KyleG.M.,Luthra,R.,etal.Aeute,short-term,and subchronic oraltoxieity1 of 1,1,l–trichloroethane in rats[J].Toxieol.Sei,2001(60):363-372
[41]周立国.药物毒理学[M].武汉:湖北科学技术出版社,2001.88-120
[42] Ames BN,Mccann J,Yamasaki E.Methods for detecting careinogens and mutagens with the Salmonella/mammalian-microsome mutagenieity test[J].MutatReS,1975(31):347一364
[43]王治乔,袁伯俊.新药临床安全性评价与实践[M].军事医学科学出版社,1997
[44] Rota PA,Obserste MS Monroe ss,etal.Characterization of a novel eoronavirus associated with severe acute respiratory syndrome[J]. Scienee,2003,300(5024):1394-1399
[45] Dayan J,Deguingand S,Truzman C,Study of the mutagenic activity of 6 hepatotoxic Pharmaeeutical drugs in the Salmonella typhimmurium microsome test,and the HGPRT and Na/K ATPase system incultured mammalian cells[J].MutatRes,1985(157):1-12
[46] Maron DM, Ames BN.Revised methods for the Sa1mon11a mutagenicity test[J].Mutation Res,1983(113):173
[47] Preston RJ,Au W,Bender, MA; et al. Mammalian in vivo and in vitro cytogenetic assays: a report of the U.S. EPA's Gene-Tox program[J].Mutat Res,1981(87):143-188
[48] Cottiglia F,Loy G,Garau D,et al.Antimicrobial evaluation of coumarins and flavonoids from the stems of Daphne gnidium L[J].phytomedicine,2001,8(4):302-305.
[49]叶和杨,马廉兰,张瑞其,等.金边瑞香的抑菌试验[J]赣南医学院学报,2002,22(5):470-471
[50] Mohanmed AH.Cytological effect of Sudium fluoride on mice[J].Fluoride, 1982,15(3): 110-112
[51] HongSK,Kitano H,Satoh H,Nagato Y.How is embryo size genetically regulated in rice[J].Development,1996,122(7):2051-2058
[52]吕晓华.体外微核试验方法研究进展[J].癌变、畸变、突变,1999,11(2):109-112
[53] Mobamly MG.Cytogenetic testing of mutagenic Potential of the blue-green bacteriam,pleetonema boryanum in experimentally treated miee[J].Cytologia,1993,58(2):161-169
[1] Curtis D Klaassen. Casarett & Doull’s Toxicology :the Basic Science of Poisons[M] . McGraw2Hill ,2001. 1-104.
[2]沈芸,陆荣柱,陈自强.应用生态化标志物评价神经毒物的效应[J] .职业卫生与应急救援,2001 ,19 (3) :124 - 125.
[3] Letz R. Continuing Challenges for Computer2based Neuropsycho2 logical Tests[J ] . Neurotoxiclogy ,2003 ,24 (4 - 5) :479 - 489.
[4] Liu B ,Gao HM ,Hong J S. Parkinson s disease and exposure to infectious agents and pesticides and the occurrence of brain injuries :role of veuroinflammation[J] . Environ Health Perspect ,2003 ,111 (8) :1065 - 1073.
[5] Rohlman DS ,Gimenes LS , Eckerman DA ,et al. Development of the Behavioral Assessment and Research System (BARS) to Detect and Characterize neurotoxicity in Humans [J ] . Neurtoxicology ,2003 ,24 (4 - 5) :523 - 531.
[6] Bellinger DC. Perspectives on incorporating human neurobehavioral end points in risk assessments [ J ] . Risk Anal ,2002 ,22 ( 3) : 487 - 498.
[7]张晓玲.浅谈中药毒副作用[J].河南中药,2002,22 (2):10—71.
[8]袁何永.由马兜铃酸致肾病谈中药对肾的毒副作用[J].广东药学,2003,13(3):5961.
[9]陈慧袱.雷公藤的毒副作用及防治[J].浙江中医学院学报,1998,22(1):46.
[10]王新红.长春新碱引起神经毒副反应l2例临床分析[J].河南神经疾病药理学杂志,2002,5(4):68.
[11]钱伟平.蟾酥对实验动物循环系统毒副作用的研究[J].浙江中医学院学报,2001,25(6):46—47.
[12]高芳.中药的毒副作用及安全用药[J].中兽医医药-47-杂志,2000,(2):43—44.
[13]胡志祥,肖金莲主编.含毒中药毒理分析[J].时珍国医国药,1999,10(2):96.
[14]陈敬炳.天然药物中的毒蛋白、多肽及氨基酸的研究概况[J].中成药,2001,23(10):751—753.
[15]张东玲,张秀英.中药毒理学的研究进展[J].中兽医医药杂志,2005,1:51
[16]孙圆嫒,高月.中药毒性的研究概况[J].航空航天医药,2003,14(4):253
[17]马利华.浅析有毒中药及毒副作用[J].时珍国医国药,2004,15(6):377.
[18]马晓彤,张翠萍,张俊刚.松花粉片的急性毒性及致突变性研究[J].医学研究与教育,2009,26(3):16-17.
[19]舒奇,傅颖媛,刘婷,余燕影.金边瑞香活性成分瑞香黄烷-Ⅰ,瑞香素对SMMC-7721和MCF-7细胞作用的研究[J].时珍国药医药,2009,20(9):2149-2150.
[20]周爱香.复方马钱子片的镇痛作用和毒理试验[J].中国实验方剂学杂志,1998,4(4):59—60.
[21] Humphries ML , Olden K.Aspraginelineked oligosaccha—fide and tumor metastatis. Pharmac[J].Ther,1989,44:85.
[22] Denms Jw,Koch K,Youselfis,eta1.Growth hibition of humaelanoma tumor xenograftesi nathymic nude mice by swainso Cancer [J].Res,1990,50:1867～1872.
[23]刘锦霞.苦豆子、铁棒锤生物碱对菜青虫的毒力测定[J].甘肃科学学报,2003,15(2):38～39.
[24]鲁小青.蓖麻毒蛋白糖脂质体对肝癌细胞的杀伤作用及其理化特性[J].上海铁道大学学报,1998,19(9):24～25.
[25]高宝岩.蓖麻毒蛋白的提取及分析[J].光谱实验室,2001,l8(4):126～127.
[26]施大钊.毒芹生物碱的提取及对害鼠的毒性实验[J].植保技术与推广,2002,22(6):27～29.
[27]马丽.毒性中药与以毒攻毒[J].现代中西医结合杂志,2001,1o(23):2324～2325.
[28]刘红云.蜂毒的研究进展及其临床应用[J].中药材,2003,26(6):457～458.
[29]张坤娟.瑞香狼毒甲醇提取物抗瘤免疫机理初探[J].中国免疫学杂志.2003,19:544.
[30]吴正杰.“十八反”“十九畏”新探—兼论中药配伍-禁忌[J].中兽医医药杂志,2001,(3):21.
[31]吴慧英.谈十九畏用于临床[J].江西中医药,1997,28(2):44.
[32] Purchase IF,Botham PA,Bmner LH,eta1. workshopoverview:scientificandregula tory challenges for the reduction,refine ment,and replacement of animals in toxic ity testing[J].Toxicol Sci,1998,43(2): 86～101.
[33] Cheek B J,Stel A B,Torres M P,eta1.Chemiluminescence detection for hybridization assays on the flow—thru chip,a three dimensional microchannel biochip[J].Anal Chem,2001,15(24):5777-5783.
[34] Pennie W D.Use of cDNA microarray to probe and understand The toxicological consequences of altered gene expression [J]. Toxicological Le tters,2000,112—113:473-474.
[35] Nuwaysir E F,Bittner M,Trent J,et a1.Microarrays and toxicology:the advent of toxicogenomics[J].Mol Carcinog,1999, 24(3):153—159.
[36] Kiela P R Midura A J,Kuscuoglu N,et a1.Efects of Boswellia serrata in mouse models of chemically induced colitis[J].Am J Physiol Gastrointest Liver Physiol,2005,288(4):G798一G808.
[37]曹煜冲药复方物质基础和药效、毒性相关性研究[J]世界科学技术-中药现代化·基础研究,1999,1(2):36.