用户名: 密码: 验证码:
平肺复方对人肺腺癌A549细胞的增殖抑制及作用机制
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
本论文包括文献综述和实验研究两个部分。
     文献综述一回顾了肺癌的中医药治疗进展,主要从病因病机、辨证分型、中医治法研究、中西医结合辨证论治、中医药维持治疗晚期肺癌、中医药治疗肺癌的机制研究六个方面进行论述。文献回顾表明中西医结合治疗肺癌注重辨证施治,标本兼顾,扶正与祛邪相结合,在改善临床症状、减轻放化疗毒副反应、放化疗增敏、提高生活质量、延长生存时间等方面取得了良好的临床疗效。
     文献综述二总结了目前对PPAR-γ信号通路的研究,主要从蛋白结构和传导途径、与肿瘤的关系、对其抗肿瘤机理研究进展等几个方面进行论述,有部分文献表明天然药物可能从PPAR-γ信号通路发挥抗肿瘤作用。
     中医药治疗肿瘤的部分机制不明,在机理方面一直缺乏突破性研究,相关信号通路与药物靶标不明,使中医药走向国际受到了一定阻碍。为探讨中药复方的可能作用靶点,本实验研究采取血清药理学、流式细胞学、蛋白质免疫印迹等方法,观察了平肺复方对人肺腺癌A549细胞的增殖抑制及作用机制。
     目的临床研究发现以养阴清肺为治法的平肺复方是延长肺癌患者生存期的独立预后因素之一,但缺乏深入研究。本实验是建立在以往相关研究的基础上,初步探讨了平肺复方生药及其含药血清对人肺腺癌A549细胞增殖的抑制作用,并观察了平肺复方对A549细胞凋亡及Ca spase-3蛋白表达水平的影响,为之后从细胞、分子角度证明平肺复方治疗肺癌的作用机理,是否影响PPAR-γ信号传导通路奠定基础,揭示中药与化疗药物治疗肿瘤的疗效差异和可能机理。
     方法
     ①平肺复方制剂20g/kg/d灌服大鼠,取含药血清培养A549细胞,设5%、15%和25%3个不同浓度正常血清组和5%、15%和25%3个不同浓度含药血清组,观察培养至0h、24h.48h.72h.96h各组细胞存活率;
     ②将平肺复方生药直接添加入含10%胎牛血清的高糖DMEM,设1mg/ml.5mg/ml和10mg/ml3个不同浓度平肺复方生药组,观察培养24h、48h、72h各组细胞存活率;
     ③将PPAR-γ特异性阻断剂GW9662与10mg/ml平肺复方生药同时作用于A549细胞培养体系,观察PPAR-γ通路阻断后平肺复方对A549细胞生长的影响;
     ④将1mg/m1.5mg/ml和10mg/ml3个不同浓度平肺复方生药作用于A549细胞,应用磷脂酰丝氨酸外翻分析法,于流式细胞仪检测细胞凋亡情况。
     ⑤将10mg/ml平肺复方生药作用于人肺腺癌A549细胞,蛋白质免疫印迹法检测培养24h.48h和72h后A549细胞中Caspase-3蛋白的表达水平,β-actin作内参。
     数据录入Excel软件进行保存,使用SPSS17.0软件进行分析,多个样本均数的比较采用t检验。
     结果
     ①在常规培养体系中添加5%-25%正常大鼠血清,对A549细胞生长存在明显影响,表现为生长抑制作用;但这种生长抑制作用与添加大鼠血清的浓度关系不大。在常规培养体系中添加25%含药大鼠血清培养至48h时即对A549细胞增殖存在影响,表现为生长抑制作用,与同浓度血清对照组的细胞存活率相比存在统计学差异(P<0.05);这种抑制作用随时间增长而增强,培养至72h、96h与对照组相比存在明显差异(P<0.01)。
     ②应用CCK-8法检测人肺腺癌A549细胞的增殖显示,平肺复方生药1mg/ml在体外作用72h内对细胞生长没有明显影响,平肺复方生药5mg/ml和10mg/ml在作用24h、48h和72h表现出对A549细胞增殖的抑制作用,与空白对照组相比存在统计学差异(P<0.05)。
     ③平肺复方生药在体外作用24h后即显现对A549细胞的生长抑制作用,这种抑制作用随时间的延长而增强,与对照组相比具有明显差异(P<0.01);单纯加入GW9662对细胞的生长并未产生影响;平肺+GW组在培养至24h、48h、72h均对细胞生长产生抑制作用,与对照组相比具有统计学差异(P<0.05),但这种抑制作用较平肺组弱(P<0.05)。
     ④平肺复方10mg/ml作用17h可引起A549细胞发生早期凋亡及死亡,与空白对照组相比存在统计学意义。
     ⑤将生药终浓度为10mg/ml的平肺复方生药作用于人肺腺癌A549细胞培养体系,可以提高肺癌细胞中Caspase-3蛋白的表达,这种作用在体外24h后即能显现,与对照组相比具有统计学意义(P<0.05);Caspase-3蛋白的表达随时间的延长有所增强,培养至72h Caspase-3蛋白的表达较24h明显增多(P<0.05)。
     结论平肺复方生药及其含药血清均可抑制人肺腺癌A549细胞的增殖;特异性拮抗剂GW9662可部分逆转平肺复方对人肺腺癌A549细胞的增殖抑制作用,平肺复方的抗肿瘤作用机制可能部分涉及到PPAR-γ信号通路,但并不完全是通过这一条信号通路在发挥作用;平肺复方可明显上调Caspase-3蛋白的表达水平,并诱导肿瘤细胞的早期凋亡。
This thesis includes two parts:two literature reviews and an experimental study.
     The first part of the literature review was the progression of the TCM therapy of lung cancer in recent years. It was including six parts:etiology and pathogenesis, syndrome differentiation, the TCM treatment research, the syndrome differentiation and treatment of integrated traditional Chinese and western medicine, the TCM maintenance therapy in advanced lung cancer, and the mechanism of Chinese medical treatment for lung cancer. This literature review showed that integration of traditional and western medicine treatment of lung cancer emphasizes the syndrome differentiation, specimens consideration, integration of strengthening healthy energy and eliminating evil. And this kind of treatment obtained satisfied clinical effect on improving clinical symptoms, reducing chemoradiotherapy adverse reaction, enhancing the quality of life, prolonging the survival time.
     The second part of the literature review summarized the research in PPAR-γ signal path at present from following several aspects:protein structure and pathway, the connection with carcinoma, the investigation progress of ant i-tumor mechanism. Some of papers showed that the natural medicine may could against the cancer cells through the PPAR-γ signal path.
     But part of mechanism of TCM therapy for cancer is still unknown. This would be likely to create difficulties on globalizing. In order to found the potential mechanism of the Pingfei Compound, this research observed the inhibitory action of the medicinal serum/apozem of the compound to the proliferation of the human lung adenocarcinoma cell line A549, and the apoptosis of the tumor cells, the expression of Caspase-3.
     Purpose This research discussed the inhibitory action of the medicinal serum/apozem of the Pingfei Compound to the proliferation of the human lung adenocarcinoma cell line A549, the apoptosis and the expression of Caspase-3, through serum pharmacology methods, flow cytometer, western blot, on the basis of previous studies.
     Methods
     ①erum containing Pingfei Compound was prepared by serologic pharmacology method. Pingfei Compound was administered orally to SD rats at doses of20g/kg/d. The proliferation of A549cells after exposure to the serum was measured by CCK-8assay at Oh,24h,48h,72h and96h.
     ②Groups were blank control, liquid extract of Pingfei prescription1mg/mL,5mg/mL and10mg/mL group. Prol iferat ion of A549cells after exposure to the apozem was measured by CCK-8assay at24h,48h, and72h.
     ③The PPAR-y specificity blocker GW9662and the Pingfei Compound were added in the cell cultures. Observe if the Pingfei Compound could influence the growth of the A549cells or not.
     ④For apoptosis study, it was detected by analyzing plasma membrane translocation of phosphatidylserine, using FACS.
     ⑤Caspase-3in A549cells was measured with Western Blot. Statistic analysis for quantitative data is student t-test.
     Results
     ⑥The growth of A549could be suppressed at the48h after exposure to the serum from rats (P<0.05). The inhibi t ion was enhanced over time. The results exists significant difference compared with the control group at the72h and96h (P<0.01)
     ⑦5mg/mL and10mg/mL groups presented inhibitive effect on A549growth at24h,48h and72h time points{P<0.01).
     ⑧GW9662reversed partly the effect of Pingfei Compound on the proliferations of A549cells.
     ⑨10mg/ml group could induse the early and late apoptosis induction (P<0.01).
     ⑩The expression of Caspase-3was significantly increased by Pingfei Compound.
     Conclusion Apozem of Pingfei Compound, a TCM formula, could suppress A549growth in vitro in dose and time dependent manner. This inhibitory effect was related to its apoptosis induction. Perhaps the increase of the expression of Caspase-3induced the apoptosis. The mechanism of Pingfei Compound may could be involved in PPAR-y signal pathway.
引文
[1]Estimated Cancer Incidence, Mortality and Prevalence Worldwide in 2012, GLOBOCAN 2012, WHO.
    [2]赫捷.陈万青.2012年中国肿瘤登记年报[M].北京:军事医学科学出版社,2012.
    [3]陈万青.张思维.曾红梅等.中国2010年恶性肿瘤发病与死亡.中国肿瘤[J],2014,23(1):1-10.
    [4]Henschke CI. Yankelevitz DF. Libby DM. et al. Survival of patients with stage I lung cancer detected on CT screening. N Engl J Med[J],2006,355:1763-1771.
    [5]NCCN Guidelines Version 3.2014 Non-small cell lung cancer.
    [6]李佩文.崔慧娟.实用中西医结合肿瘤内科学.中国中医药出版社[M],2007:18.
    [7]Robert A. Gatenby. A change of strategy in the war on cancer. Nature[J],2009,459 (7246):508-509.
    [8]薛青.尤建良.肺癌中医药诊疗现状分析.辽宁中医药大学学报[J],2012,14(8):260-262.
    [9]崔慧娟.张培宇.张代钊教授治疗肺癌经验.中日友好医院学报[J],2011,25(1):57-58.
    [10]郑红刚.花宝金.朴炳奎辨治肺癌学术思想与经验探析.中医杂志[J],2010,51(4):304-306.
    [11]郑红刚.花宝金.朴炳奎辨证治疗肺癌的学术思想.北京中医[J],2007,26(5):273-275.
    [12]张霆.从伏气学说探讨肺癌之病因病机.中医研究[J],2007,20(3):5-7.
    [13]张霆.肺癌转移病机探析.山西中医[J],2007,23(1):76-77.
    [14]唐引引.徐立然.原发性支气管肺癌中医证型和用药规律分析.中医学报[J],2012,27(166):278-280.
    [15]李丛煌.花宝金.283例中晚期非小细胞肺癌患者证候分布及证候要素组合特点分析.北京中医药大学学报[J],2009,32(10):712-715.
    [16]黎敬波.张征.林丽珠等.非小细胞肺癌417例证候分布的临床研究.广州中医药大学学报[J],2009,26(1):73-77.
    [17]游捷.李泰峰.汪霞等.303例老年原发性支气管肺癌中医证候分型.中医杂志[J],2012,53(16):1404-1407.
    [18]王少墨.董志毅等.388例原发性肺癌中医证候分布状况分析.上海中医药大学学报[J],2013,27(5):30-34.
    [19]黄东华.周俊琴.鲁琴等.140例中晚期肺癌患者中医证型分布规律研究.新中医[J],2013,45(6):105-106.
    [20]谢长生.王东建.潘磊等.561例肺癌中医证型与TNM分期及病理类型的相关探索.浙江中医杂志[J],2010,45(6):398-399.
    [21]张明萍.王建军.李志宏.中晚期非小细胞肺癌中医证型的临床观察.世界中西医结合杂志[J],7(8):706-708.
    [22]尤杰.刘嘉湘教授辨治肺癌经验.长春中医药大学学报[J],2011,27(2):185-186.
    [23]吴继.刘嘉湘扶正治疗肺癌用药经验.辽宁中医杂志[J],2012,39(4):617-619.
    [24]王雄文.周蓓.周岱翰教授应用温补法治疗肺癌经验介绍.新中医[J],2009,41(6):11-12.
    [25]兰智慧.朱良春辨治肺癌经验.上海中医药杂志[J],2010,44(9):1-2.
    [26]李际强.刘伟胜攻毒法治疗肺癌经验简介.辽宁中医杂志[J],2009,36(3):340-341.
    [27]田静.丁金芳.施志明教授用虫类药治疗肺癌经验介绍.新中医[J],2004,36(3):12-13.
    [28]程海波.吴勉华.周红光.周仲瑛从癌毒辨治恶性肿瘤的经验.北京中医药[J],2009,28(11):844-846.
    [29]程海波.吴勉华.周仲瑛教授“癌毒”学术思想探析.中华中医药杂志[J],25(6):866-869.
    [30]张霆.肺癌治痰刍议.中医药临床杂志[J],2007,19(1):77-78.
    [31]张霆.肺癌治痰使用药对分析[J].陕西中医,2007,28(4):459-460.
    [32]张霆.张建勇.肺癌分期治痰探析.中医药临床杂志[J],2007,19(3):299-300.
    [33]郭海涛.温阳化痰法治疗肺癌刍议[J].光明中医,2010,25(4):680-681.
    [34]李麒.陈泽涛.活血化瘀法改善原发性支气管肺癌高凝状态的临床观察.辽宁中医药大学学报[J],15(8):182-185.
    [35]陈永东.王远东.邵中夫.康莱特联合新辅助化疗对ⅢA期非小细胞肺癌术后并发症的临床观察.现代肿瘤医学[J],2004,12(4):326-327.
    [36]覃国亮.冼磊.陈铭伍.参芪扶正注射液对肺癌患者围术期免疫功能的影响.广西医学[J].2013,35(2):170-171.
    [37]蒋泽华.王达飞.陈胜东等.参冬犀黄汤配合西药治疗放射性肺炎82例.南京中医药大学学报[J],2011,27(6):533-535.
    [38]陈连生.杨光华.付尚志等.银翘温胆汤治疗放射性肺病180例临床疗效观察.临床军医杂志[J],2013,41(10):1010-1012.
    [39]何萍.袁红.邵艳欣等.益气化淤汤对Ⅲ其非小细胞肺癌放疗患者生存质量的临床观察.新中医[J],2013,45(4):98-99.
    [40]黄金昶.张代钊.李佩文等.扶正增效方配合肺癌放疗远期疗效分析.中国医刊[J],2000,35(1):48.
    [41]尹晓东.吴晓静.牛静秀等.中药联合放射治疗非小细胞肺癌的观察.中国中西医结合外科杂志[J],2013,19(6):621-623.
    [42]孙建立.刘嘉湘.李春杰等.扶正法辨证结合化疗治疗晚期非小细胞肺癌近期疗效和对生活质量影响的临床研究.辽宁中医杂志[J],38(6):1039-1041.
    [43]盛晶.吉西他滨配合中药治疗老年晚期非小细胞肺癌的疗效观察.中国实用医药[J],2011,6(11):175-176.
    [44]韩承镇.扶正增效方联合化疗治疗晚期非小细胞肺癌对照观察.实用中医内科杂志[J],2012,26(8):26-27.
    [45]王莉.张祺箐.冯正权.中医药对中晚期非小细胞肺癌生存期的影响.云南中医学院学报[J],2013,36(4):47-49.
    [46]李平.中医辨证配合化疗治疗晚期肺癌30例观察.中医药临床杂志[J],2009,21(6):529.
    [47]朱丽华.李和根.史美育等.非小细胞肺癌根治术后无瘤生存期影响因素分析及中药干预效果评价.上海中医药杂志[J],2013,47(2):11-15.
    [48]李秀荣.周延峰.焦中华等.消瘤平移合剂抗肿瘤术后转移的临床观察.中国中西医结合杂志[J],2001,21(5):335-338.
    [49]李秀荣.齐元富.周延峰等平移合剂抗肿瘤术后转移的临床研究.中国中医药信息杂志[J],2008,15(10),9-11.
    [50]王中奇.徐振晔.邓海滨等.中医药联合化疗防治非小细胞肺癌术后复发转移的临床研究.上海中医药杂志[J],2011,45(5):36-39.
    [51]张志娣.黄挺.杨少山等.益肺颗粒联合化疗预防肺癌术后转移疗效观察.中医药学刊[J],2005,23(4):643-644.
    [52]杨国良.中晚期非小细胞肺癌中医证型与吉非替尼治疗敏感性关系研究.辽宁中医杂志[J],2009,,36(6):867-869.
    [53]孙建立.中医辨证结合吉非替尼治疗晚期非小细胞肺癌临床疗效及证候变化分析.四川中医[J],2009,,27(11):64-66.
    [54]吴万垠.杨小兵.杨昌卫等.中药联合吉非替尼一线治疗晚期非小细胞肺癌[C],第三届国际中医、中西医结合肿瘤学术交流大会暨第十二届全国中西医结合肿瘤学术大会论文汇编.浙江宁波.2010.
    [55]张琦.袁海.康莱特注射液联合吉非替尼治疗晚期非小细胞肺癌78例临床观察.肿瘤[J],2011,31(1):89-90.
    [56]梁翠微.杨兵.杜均祥等.吉非替尼相关皮疹的中医辨证论治.中国实用医药[J], 2011,6(16):22-23.
    [57]吴益萍.凉血消风散加减治疗吉非替尼相关皮疹临床观察.新中医[J],2013,45(2):53-54.
    [58]柴小姝.何春霞.吴万垠等.消积饮维持治疗晚期非小细胞肺癌对生存期的影响.时珍国医国药[J],2011,22(10):2547-2548.
    [59]曾冬香.毕延智.盛桂凤等.参一胶囊在晚期非小细胞肺癌维持治疗中的疗效观察.癌症进展[J],2013,11(6):554-557.
    [60]王璐.孙智霞.冯光强等.中医辨证维持治疗对晚期非小细胞肺癌化疗后生存质量及无进展生存期的影响.中国实验方剂学杂志[J],2013,19(13):319-322.
    [61]潘玉真.殷东风.周立江等.柴胡龙牡汤对Lewis肺癌小鼠细胞免疫功能的影响.临床肿瘤学杂志[J],2011,16(3):211-215.
    [62]郭净.王菊勇.郑展等.肺炎宁方对肺癌小鼠CD4+ CD25+调节性T细胞比例及Foxp3表达的影响.中西医结合学报[J],2012,10(5):584-590.
    [63]杨国良.张学进.胡丹丹.肺积方联合化疗对Lewis肺癌移植小鼠免疫逃逸相关细胞因子及CD4+ CD25+ Treg细胞的影响.中华中医药学刊[J],2012,,30(12):2713-2716.
    [64]郑璐玉.熊飞.詹臻等.麦门冬汤合千金苇茎汤提取部位对非小细胞肺癌H460细胞毒作用的研究,中国实验方剂学杂志[J],2010,16(3):60-63.
    [65]胡家才.李清泉.川芎嗪对肺癌A549细胞增殖与凋亡的影响及其机制.武汉大学学报(医学版)[J],2010,31(1):19-21.
    [66]张涛.陆素青.欧阳新平等.冬凌草甲素对体外培养的前列腺癌PC-3细胞生长的影响.中国医学工程[J],2012,20(10):160-161.
    [67]Folkman J. Tumor angiogenesis:therapeutic implications [J]. N Engl J Med, 1971,285(21):1182-1186.
    [68]陈培丰.潘磊.金莹祺.清热消积方对人肺腺癌细胞诱导的人脐静脉内皮细胞迁移、趋化和成管能力的影响.中国中西医结合杂志[J],2013,33(4):497-501.
    [69]王中奇.邓海滨.张丽曼等.中药肺岩宁方及其拆方对小鼠Lewis肿瘤生长、肺转移和微血管密度的影响[J].中华中医药杂志,2012,27(2):477-480.
    [70]赵连梅.孙佳玮.颜晰等.中药升陷汤抑制肺癌A549细胞增殖和侵袭转移作用研究.中华中医药杂志[J],2011,26(9):2147-2150.
    [71]赵晶.姜达.恶性肿瘤高凝状态及其干预.癌症进展[J],2013,11(1):48-52.
    [72]Farge D. Durant C. Villiers S, et al. Lessons from French National Guidelines on the treatment of venous thrombosis and central venous catheter thrombosis in cancer patients. Thromb Res [J],2010,125 (suppl 2):s108-116.
    [73]Noble S. Pasi J. Epidemiology and pathophysiology of cancer-associated thrombosis. Br J Cancer [J],2010,102:s2-29.
    [74]胡艳.侯安继.周莉等.益气活血方对中晚期恶性肿瘤高凝状态的影响.上海中医药杂志[J],2011,45(8):36-38.
    [75]毛德奎.秦海峰.于爱平等.肺癌患者化疗前后凝血、纤溶功能与血小板参数的变化及其临床意义.军事医学[J],2012,36(2):141-146.
    [76]李麒.陈泽涛.活血化瘀法改善原发性支气管肺癌高凝状态临床观察.辽宁中医药[J],2013,15(8):182-185.
    [77]于明薇.杨国旺.王笑民.化瘀丸对小鼠Lewis肺癌生长及血栓形成相关因子的干预作用.中国实验方剂学杂志[J],2011,17(17):163-166.
    [78]Xuguang Zhang. Xiangwen Peng. Weiping Yu, et al. Alpha-tocopheryl succinate enhances doxorubicin-induced apoptosis in human gastric cancer cells via promotion of doxorubicin influx and suppression of doxorubicin efflux. Cancer Letter [J],2011,307(2):174-181.
    [79]农丽.伍钢.戴晓芳等.吴茱萸碱逆转人肺癌细胞株A549/DDP耐药机理的实验研究.临床肿瘤学杂志[J].2010,15(6):487-492.
    [80]张莹.贾英杰.杨洁等.消岩汤药物血清对A549/DDP多药耐药逆转作用的研究.天津中医院[J].2010.27(4):334-336.
    [81]吴智春.于华芸.季旭明等.温下方联合顺铂对A549/DDP裸鼠移植瘤的抑制作用及对药物转运蛋白表达的影响[J].中华临床医师杂志:电子版,2011,5(20):5884-5888.
    [1]Boitier E, Gautier JC, Roberts R. Advances in understanding the regulation of apoptosis and mitosis by peroxisome-proliferator activated receptors in pre-clinical models:relevance for human health and disease. Comp Hepatol 2003, 2 (1):3.
    [2]Issemam I. Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature[J],1990,347:645-650.
    [3]Willson TM, Brown PJ, Strenbach DD, et al. The PPARs:From orphan receptors tu drug discovery, J Med Chem[J],2000,43:527-550.
    [4]Berger J, Moller DE. The mechanisms of action of PPARs. Ann Rev Med[J],2002,53:409-435.
    [5]Sher T, Yi H F, McBride OW, et al. cDNA cloning, chromosomal mapping, and functional characterization of the human peroxisome proliferator activated receptor. Biochemistry 1993,32,5598-5604.
    [6]MarxN, Duez H, Fruchart JC, et al. Peroxisome proliferator-activated receptors and atherogenesis:regulators of gene exp ression in vascular cells [J]. Circ Res,2004,94 (9):1168-1178.
    [7]Sheu SH, Kaya T, Waxman DJ, Vajda S: Exploring the binding site structure of the PPAR gamma ligand-binding domain by computational solvent mapping. Biochemistry (Mosc) 2005,44 (4):1193-1209.
    [8]Lehrke M, Lazar MA: The many faces of PPARgamma. Cell 2005,123 (6):993-999.
    [9]Feige JN, Gelman L, Michalik L, Desvergne B, Wahli W: From molecular action to physiological outputs:peroxisome proliferator-activated receptors are nuclearreceptors at the crossroads of key cellular functions. Prog Lipid Res 2006, 45(2):120-159.
    [10]Margeli A, Kouraklis G, Theocharis S: Peroxisome proliferator activated receptor-gamma (PPAR-gamma) ligands and angiogenesis. Angiogenesis 2003,6(3):165-169.
    [11]Panigrahy D, Huang S, Kieran MW, et al. PPARgamma as a therapeutic target for tumor angiogenesis and metastasis. Cancer Biol Ther 2005,4(7):687-693.
    [12]Greene ME. Blumberg B. McBride OW, et al. Isolation of the human peroxisome proliferator activated receptor gamma cDNA: expression in hematopoietic cells and chromosomal mapping. Gene Expression 1995,4,281-299.
    [13]Fajas L. Auboeuf, D.; Raspe, E. et al. The organization, promoter analysis, and expression of the human PPARy gene. J. Biol. Chem. 1997,272,18779-18789.
    [14]Owen G. I. and Zelent A. (2000):Origins and evolutionary diversification of nuclear receptor superfamily. CMLS Cell. Mol. Life Sci.,57,809-827.
    [15]Werman A., Hollenberg A., Solanes G., et al. (1997):Ligand-independent activation domain in the N terminus of peroxisome proliferator-activated receptor gamma (PPARy). J. Biol. Chem.,272,20230-20235.
    [16]Shao D., Rangwala S. M., Bailey S. et al. (1998):Interdomain communication regulating ligand binding by PPAR-gamma. Nature, 396,377-380.
    [17]Nolte RT, Wisely GB, Westin S, et al. Ligand binding and co-activator assembly of the peroxisome proliferators activated receptor-y Nature[J],1998,395:137-143.
    [18]Fajas L, Auboeuf D, Raspe E, et al. The organization, promoter analysis and expression of the human PPAR-y gene. J Biol Chem[J],1997,272:1877-1878.
    [19]Feige JN, Gelman L, Michalik L, et al. From: molecular action to physiological outputs:Peroxisome proliferator-activated receptors are nuclear receptors at the crossroads of key cellular functions. Prog Lipid Res [J],2006,45:120-159.
    [20]Lehmann, J. M.; Moore, L. B.; Smith-Oliver, T. A. et al. An antidiabetic thiazolidinedioneis a high affinity ligand for peroxisome proliferator-activated receptor c (PPARc). J. Biol. Chem. 1995,270, 12953-12956.
    [21]HaroldMW, Clary BC. A synthetic antagonist for the peroxisome proliferator-activited receptory inhibits adipocyte differentiation [J]. J Biol Chem,2000; 275:1873-1877.
    [22]Rieusset J, Touri F. A new selective peroxisome proliferators-activated receptor y antagonist with antibosity and antidiabetic activity[J]. Mol Endocrinol,2002; 16:2628-2644.
    [23]余晶晶.朱小萍.张伟.PPAR-γ在非小细胞肺癌中的表达及临床相关性研究.实用临床医学[J],2006,7(12):5-8.
    [24]何晓燕,张敏,陈智超,等.PPAR-γ在肺癌中的表达及在肺癌凋亡中的作用研究[J].中国肺癌杂志,2006,9(1):35-39
    [25]ZHANG Min. YOU Yong. ZOU Ping. Et al. Expression of PPAR-y in Lung Cancer and the Role of PPAR-y in Apoptosis of Lung Cancer. The Chinese-German Journal of Clinical Oncology [J],2005,4(1):36-39.
    [26]李虹.林箐.宋琳琳等.PPAR-γ在非小细胞肺癌中的表达及其在预后评估中的意义[J],2012,22(5):35-38.
    [27]Giaginis C. Politi E. Alexandrou P. et al. Expression of Perosisome Proliferator activated receptor-gamma in human non-small cell lung carcinoma: correlation with clinicopathological parameters, proliferation and apoptosis related moleculs and patients'survival. Pathol.Oncol. Res. [J],2012,18:875-883.
    [28]李强.党诚学.胃癌组织中PPAR-γ的表达.山东医药[J],2011,51(34):40-41.
    [29]李强.党诚学. PPAR-γ mRNA在胃癌组织中的表达及其与胃癌预后的关系.实用癌症杂志[J],2012,27(1):18-20.
    [30]刘鹏飞.刘顺英.钱建忠等.PPAR-γ和MMP-7与胃癌及癌前病变的关系.中华消化内镜杂志[J],2005,22(3):171-173.
    [31]马秀梅.左连富.刘江惠等.人胃癌组织中PPAR-γ的表达及其配体对胃癌细胞生长的影响.基础医学与临床[J],2006,26(3):294-301.
    [32]吴旭东.陈卫昌.张素青.PPAR-γ在肝细胞癌组织中的表达及其临床意义.江苏医药[J],2007,33(11):1106-1109.
    [33]张凤新.彭利.张萌等.PPAR-γ和p27、Skp2在肝细胞癌中的表达及临床意义.天津医药[J],2012,40(7):654-662.
    [34]何坤.胡泽民.周载平等.过氧化物酶体增殖物激活受体γ在肝癌组织中的表达及意义.肿瘤防治研究[J],2009,36(12):1047-1053.
    [35]沈丹.邓长生.过氧化物酶体增殖物激活受体γ在结肠癌肿的表达及其激动剂对结肠癌细胞的生长抑制作用.临床内科杂志[J],2008,25(1):57-60.
    [36]李慧艳.谢贤和.侯梅.PPAR-γ在结直肠癌中的表达及临床意义.海南医学[J],2011,22(12):12-14.
    [37]舒振波.操海萍.盖宝东等.PPAR在结直肠癌中的表达及意义.中国实验诊断学[J],2008,122(3):327-329.
    [38]陈功波.夏杰.曹杰等.PPAR-γ、urvivin在直肠癌中的表达及其临床意义.中国医学工程[J],2009,17(2):85-87.
    [39]何英.邬玉辉.乳腺癌PPAR-γ和 COX-2的表达及临床意义.中国现代医学杂志[J],2011,21(20):2421-2424.
    [40]Zhang GY,Ahmed N, Riley C. et al. Enhanced expression of peroxisome proliferator-activated receptor gamma in epithelial ovarian carcinoma [J]. Br
    J Cancer. 2005; 92 (1):113-119.
    [41]任黔川.谭欣.彭芝兰等.PPAR-Y在上皮性卵巢癌的表达及其意义.中国现代医学杂志[J],2009,19(21):3286-3289.
    [42]孔蕾.张家文.于甬华等.PPAR-γ、PTEN. P27在子宫内膜癌中的表达及其相关性研究.四川大学学报(医学版)[J],2007,38(2):334-336.
    [43]陆谔梅.叶劲军.黄健等.PPAR-γ和 P27在官颈癌组织中的表达及其临床意义.临床肿瘤学杂志[J],2011,16(5):433-436.
    [44]梁少强.涂青松.黄荣等.宫颈癌中PPAR-γ的表达及其意义.中国医药科学[J],2012,2(15):18-19,25.
    [45]祝海.翁博文.徐珞等.前列腺癌组织PPAR-γ及其配体表达和作用机制的研究.中华肿瘤防治杂志[J],2012,19(11):840-843.
    [46]娄金峰.牛光明.娄季宇等.PPAR-γ在侵袭性垂体瘤中的表达.中国实用神经疾病杂志[J],2010,13(24):30-32.
    [47]卢瑶.罗佐杰.秦映芬等.肾上腺皮质肿瘤中PPAR-γ与 PTEN蛋白的表达及相互关系.第二军医大学学报[J],2008,29(11):1398-1399.
    [48]赵颖海.黄坊.景志亮.鼻咽癌细胞中PPAR-γ、p-ERK、cyclinD1、Ki-67蛋白的表达及意义.临床与实验病理学杂志[J],2010,26(2):196-199.
    [49]林裕茂.张乃崇.赖廷海等.PPAR-γ和Kiss-1在脑胶质瘤的表达及其与预后的关系.广东医学[J],2011,32(12):1584-1585.
    [50]沈波.聂玉强.过氧化物酶体增殖物激活受体Y抑制肝癌细胞的增殖生长和侵袭转移.临床肝胆病杂志[J],2013,29(9):702-706.
    [51]张克惠.伍仕敏.杨华芬等.PPAR-γ依赖和非依赖途径参与吡格列酮对人肝癌细胞的增殖抑制作用.军事医学[J],2011,35(7):521-526.
    [52]朱理辉.张琍等.PPAR-γ激活剂罗格列酮对人胃癌MGC803细胞周期及其PTEN表达的影响.实用癌症杂志[J],2006,21(3):238-240.
    [53]Takeuchi S, Okumura T, Mot omura W, et al. Troglitazone induces G1 arrest by p27 (Kipl) induction that is mediated by inhibition of proteasome in human gastric cancer cells[J]. Jpn J Cancer Res, 2002,93(7):774.
    [54]Han S, Sidell N, Fisher PB, et al. Up-regulation of p21 gene expression by peroxisome proliferators activated receptor gamma in human lung carcinoma cells. Clin Cancer Res [J],2004; 10(6):1911.
    [55]欧阳义.过氧化物酶体增殖物激活受体对结肠癌细胞增殖与凋亡的影响.中外医学研究[J],2011,9(9):35-36.
    [56]张萌.彭利.乔治斌等.罗格列酮激活P38MAPK通路调控P53及P21影响人肝癌HepG2 细胞周期.第三军医大学学报[J],2013,12.(网络优先出版)
    [57]张敏.邹萍.白明等.活化的过氧化物酶体增生物激活受体-Y诱导人类肺癌细胞凋亡及其机制的研究.中华医学杂志[J],2003,83(13):1169-1172.
    [58]翟福林.庄英帜.曹建国.罗格列酮下调肺腺癌细胞血管内皮细胞生长因子的表达.中国药理学通报.2006,22(10):1254-1258.
    [59]李敏.程南生.熊先泽等.PPAR-γ配体吡格列酮对胆管癌细胞体外侵袭力的影响及其机制.中国现代医学杂志[J],2007,17(16):1962-1965.
    [60]陈宏辉.张琍.刘灿等.罗格列酮对人胃癌细胞系迁移侵袭的抑制及其机制.世界华人消化杂志[J].2011,19(21):2207-2213.
    [61]B. Shen, ESH Chu, G Zhao, et al. PPARgamma inhibits hepatocellular carcinoma metastases in vitro and in mice. British Journal of Cancer [J].2012,106:1486-1494.
    [62]张涛.周洁.陈敏等.姜黄素通过PPAR-γ途径促进人肾癌细胞的凋亡.现代泌尿外科杂志[J],2013,1(18):13-18.
    [63]骆殊.沈洪.朱学军等.黄芪、莪术配伍方对胃癌COX-2、PPAR-γ及转移相关因子表达的影响.药学与临床研究[J],2009,17(1):14-17.
    [64]Kristiansen G, Jacob J, Buckendahl A C, Grutzmann R, Al ldinger I, Sipos B, et al. Peroxisome proliferator-activated receptor gamma is highly expressed in pancreatic cancer and is associated with shorter overall survival times [J]. Cl in Cancer Res,2006,12:6444-6451.
    [65]H.Li, M. C. M. Weiser-Evans, R. Nemenoff, Anti-and Protumorigenic effects of PPAR-γ in lung cancer progression: a double-edged sword. PPAR Research[J], 2012,1-12.
    [1]唐引引.徐立然.原发性支气管肺癌中医证型和用药规律分析.中医学报[J],2012,27(166):278-280.
    [2]王少墨.董志毅等.388例原发性肺癌中医证候分布状况分析.上海中医药大学学报[J],2013,27(5):30-34.
    [3]蔡光蓉.王雪玉.李佩文滋阴润肺法在肺癌治疗中的应用.中国中西医结合外科杂志[J],2001,7(3):179.
    [4]杨宪勇.北沙参对免疫抑制C57BL/6J小鼠T淋巴细胞亚群影响的实验研究.泰山医学院学报[J],2012,33(4):247-249.
    [5]姚岚,盛丽,王莉,等.沙参对肺纤维化大鼠FN、LN的影响.中国工业医学杂志[J],2007,20(2):118-119.
    [6]金香男,郑明昱.北沙参乙醇提取物对四氯化碳诱导急性肝损伤的保护作用.长春中医药大学学报[J],2010,26(6):828-829.
    [7]刘西岭,辛华,谭玲玲,等.北沙参水提法不同提取物体外抗肿瘤的研究. 安徽农业科学[J],2009,37(20):9481-9482,9490.
    [8]冯佩佩.李忠祥.原忠.党参属药用植物化学成分和药理研究进展.沈阳药科大学学报[J],2012,29(4):307-311.
    [9]柏长青.宋颖芳.王德堂等.黄芪、党参提取物抑制肺癌细胞诱导血管内皮细胞迁移的实验研究.武警医学[J],2008,19(6):505-508.
    [10]弥曼.李汾.任利君等.百合多糖的分离纯化及抗肿瘤作用.西安交通大学学报(医学版)[J],2009,30(2):177-180.
    [11]何纯莲,杨小红,黄浩,等.百合多糖的抗疲劳作用.湖南师范大学学报:医学版[J],2009,6(3):9.
    [12]唐明.百合多糖的提取、纯化及抑菌活性研究[D].长沙:湖南农业大学,2010.
    [13]许秋菊.侯莉莉.胡国强等.麦冬皂苷B诱导人宫颈癌HeLa细胞自噬的机制.药学学报[J],2013,48(6):855-859.
    [14]Min HY, Park EJ, Hong JY, et al. Antiproliferative effects of dibenzocyclooctadiene lignans isolated from Schisandra chinensis in human cancer cells. Bioorg Med Chem Lett [J],2008,18 (2):523-6.
    [15]Nishida H, Tatewaki N, Nakajima Y, et al. Inhibition of ATR protein kinase activity by schisandrin B in DNA damage response. Nucleic Acids Res [J], 2009, 37 (17):5678-89.
    [16]Oh EY, Jang JY,Choi YH, et al. Inhibitory effect of 1-0-methyl-fructofuranose from Schisandra chinensis fruit on melanogenesis in B16F0 melanoma cells. J Ethnopharmacol [J],2010,132 (1):219-24.
    [17]Li L, Fan QX. Reversal effect of Schisandrin A on MRPl-mediated multidrug resistance. J Chin Pract Diagn Ther [J], 2009,23 (9):857-9.
    [18]Sun M, Xu X, Lu QH, et al. Schisandrin B: a dual inhibitor of Pglycoprotein and multidrug resistance-associated protein. Cancer Lett [J],2007 246(1-2):300-7.
    [19]张明发.沈雅琴.桑白皮的药理研究进展.上海医药[J],2006,27(4):164-167.
    [20]邹丽宜.陈忻.吴铁等.桑白皮低聚壳聚糖体内抗肿瘤作用研究.现代中西医结合杂志[J],2007,16(1):28-29.
    [21]周颖,季晖,李萍,等.五种贝母甾体生物碱对豚鼠离体气管条M受体的拮抗作用.中国药科大学学报[J],2003,34(1):58-60.
    [22]李仝,胡凯文,陈信义,等.浙贝母对呼吸系统耐药金黄色葡萄球菌逆转作用的临床研究.北京中医药大学学报[J],2001,24(5):51-52.
    [23]唐晓勇.浙贝母碱逆转肺癌A549/DDP细胞株多药耐药及其机理研究[D].山东中医药大学,2012.
    [24]张霄翔,王艳苹,王玉凤等.瓜蒌皮对环磷酰胺致免疫功能低下小鼠免疫功能的影响[J].中国药房,2009,20(9):648-650.
    [25]秦林,高伟良.瓜蒌对子宫颈癌细胞和巨噬细胞的影响[J].山东中医学院学报,1995,19(6):414-416.
    [26]陆波,徐亚敏,张汉明,等.白芨不同提取部位对家兔血小板聚集的影响.解放军药学学报[J],2005,21(5):330-332.
    [27]刘明志,唐建洲,张建社,等.白芨中萜类化合物对人脐静脉内皮细胞凋亡和细胞骨架的作用.生命科学研究[J],2009,13(6):482-486.
    [28]张颖.周歧新.赖舒.白芨多糖对骨髓造血功能的影响.中药药理与临床[J],2009,25(4):35-37.
    [29]薛兴阳.付腾飞.邵方元等.鱼腥草总黄酮对人肿瘤细胞的抗肿瘤活性作用.现代中西医结合杂志[J],2013,22(23):2509-2511.
    [30]李涛.余旭亚.韩本勇.白花蛇舌草抑菌作用研究.时珍国医国药[J],2008,19(6):1335-6.
    [31]王宇翎.张艳.方明等.白花蛇舌草总黄酮的免疫调节作用.中国药理学通报[J],2005,21(4):444-7.
    [32]曾永长.梁少瑜.罗佳波等.白花蛇舌草提取物(HDNF)体内外抗肿瘤实验研究.中药 材[J],2011,34(4):594-597.
    [33]李洁.李玉基.李博.白花蛇舌草抑制Lewis肺癌小鼠自发转移的实验研究.中医药导报[J],2012,18(9):78-82.
    [34]朱世杰.李佩文.贾立群.平肺口服液对荷瘤小鼠细胞增殖的双向调节作用.中西医结合学报[J],2003,1(3):202-204.
    [35]朱世杰.李佩文.贾立群.养阴清肺方治疗肺癌的机理研究.北京中医药大学学报[J],2004,2(27):64-67.
    [36]程志强.张嘉.刘轩等.平肺口服液在非小细胞肺癌放疗中的应用.中日友好医院学报[J],2009,23(6):334-336.
    [37]刘轩.李红艳.夏启胜等.平肺口服液抑制肺成纤维细胞生长和诱导凋亡.中药新药与临床药理[J],2010,21(2):127-130.
    [38]许灿龙.养阴清肺法治疗肺癌的PPAR-γ信号通路研究.2010年北京中医药大学硕士学位论文.
    [39]崔海兰.李佩文.贾立群.平肺口服液对肺癌恶病质小鼠的免疫调节作用.中华中医药学刊[J],2007,25(4):714-715.
    [40]崔海兰.李佩文.贾立群.平肺口服液调节肺癌小鼠血清瘦素水平的实验研究.中日友好医院学报[J],2006,20(4):222-224.
    [41]Iwama H. Amagaya S. Ogihara Y. et al.Effect of shosaikoto a Japanese and Chinese traditional herbal medical mixture on the mitogentic activity of lipopolysaccharide: a new pharmacological testing model, J Ethonpharmacol [J], 1987,21 (1):45.
    [43]潘卫松.刘美风.石钺等.血清药理学、血清化学和中药药代动力学.世界科学技术一中药现代化[J],2002,4(3):53-55.
    [44]葛金文,朱惠斌,王宇红等.关于中药血清药理学方法的再思考[J].世界科学技术——中医药现代化,2008,10(6):16.
    [45]李平,楚更五,周华等.蜂胶左金丸含药血清抗幽门螺杆菌的实验研究[J].中国实验方剂学杂志,2008,14(3):59.
    [46]韩俭,于红娟,吴勇杰等.中药血清药理学方法学研究——抗菌试验含药血清处理方案的研究[J].中药药理与临床,2002,18(1):47.
    [47]周明眉,杨奎,姜远平等.中药血清药理学的方法学研究——含药血清低温保存和血清灭活的影响[J].中药药理与临床,1999,15(2):44.
    [48]许炜茹.林洪生.陈信义等.中药复方体外药理研究的思考.中华中医药学刊[J], 2011,,29(1):55-56.
    [49]金惠铭.王建枝.王万铁等.病理生理学[M].2008.
    [50]孙曼.蒋军广.张晓燕等.凋亡相关基因caspase-3在非小细胞肺癌中的表达及临床意义.中国老年学杂志[J],2014,34(1):17-19.
    [51]赵阳.李晓军.隋昕等PCNA和caspa se-3在肺癌组织中的表达及意义.细胞与分子免疫学杂志[J],2010,26(2):154-156.
    [52]赵雅瑞.张立凡.刘特.姜黄素上调caspase-3和P53表达后对人肺癌干细胞增殖与侵袭的影响.中国临床医学[J],2013,20(3):259-262.
    [53]李秀荣.李慧杰.王秀娟.六神丸对肺癌A549细胞caspase-3及Survivin表达的影响.中医学报[J],2013,28(179):469-471.
    [54]柴可群.赵同伟.卢丽琴等.中药抑肺饮对肺腺癌细胞A549裸鼠移植瘤的抑瘤作用及相关机制研究.中华中医药杂志[J],2010,25(3),442-446.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700