健脾疏肝解毒方对大鼠肝癌P53，survivin基因表达影响的研究
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摘要
目的:探讨中药健脾疏肝解毒方对DEN诱发性大鼠肝癌的作用和机制,研究脾虚肝郁刺激对肝癌形成的影响。以期为临床运用健脾疏肝解毒方治疗肝癌特别是预防和控制肝癌形成提供客观的依据。
方法:健康72只SD雄性大鼠,体重110士20g,正常饲养三天,称重后按SPSS随机分为DEN对照组,DEN中药高剂量组,DEN中药低剂量组;病证结合模型组,病证结合中药高剂量组,病证结合中药低剂量组。DEN水溶液按照浓度为1mlDEN:10000ml灭菌蒸馏水的比例配成,给予分组后各组大鼠自由饮用,每天饮用,连续22周,建立诱发性肝癌大鼠实验模型。病证结合模型各组大鼠采用先连续灌胃生大黄煎液,并慢性束缚大鼠四肢,限制活动和断食以施加脾虚肝郁刺激因子,同时给予DEN溶液自由饮用,建立脾虚肝郁大鼠肝癌病证模型。DEN中药高、低剂量组和病证结合中药高、低剂量组,分别按30mg/kg、10mg/kg给予中药,DEN对照组和病证模型组给予生理盐水,连续22周。实验结束后第2天,各组大鼠于乙醚麻醉下处死,观察肝脏表面病变结节,称取肝脏、脾脏重量,计算肝脏指数,脾指数。HE染色,光镜下观察大鼠肝脏组织结构,并运用免疫组化法检测肝脏组织突变型P53,Survivin的表达水平。
结果:大鼠经脾虚肝郁因素刺激和DEN作用下的病证结合模型组,与单纯DEN诱发性大鼠肝癌对照组比较,免疫器官的重量有显著下降,二者有显著性差异(P＜0.05);而健脾疏肝解毒方能显著提高诱癌大鼠免疫器官的重量,各组大鼠肝指数、脾指数结果与DEN对照组相比,DEN中药高、低剂量组有显著统计学意义(P＜0.01),中药高剂量组脾指数显著高于中药低剂量组(P＜0.01);中药高、低剂量组肝指数低于DEN对照组(P＜0.01);病证中药高、低剂量组脾指数均高于病证模型组,尤其是病证中药高剂量组更为显著(P＜0.01);病证中药高、低剂量组肝指数均低于病证模型组(P＜0.01)。健脾疏肝解毒方亦能降低肝癌组织突变型P53,Survivin蛋白的表达量,与DEN对照组相比,DEN中药高、低剂量组有明显差异(P＜0.01);与病证结合模型组相比,病证中药高、低剂量组均有统计学意义(P＜0.05),病证结合中药高剂量组差异显著(P＜0.01)。
结论:健脾疏肝解毒方能明显抑制大鼠肝癌的形成,并能增强肝癌大鼠免疫器官的重量,其作用机制可能与健脾疏肝解毒方下调肝癌组织突变P53,Survivin基因表达以及增强肝癌大鼠的免疫功能有关。并且脾虚肝郁刺激对二乙基亚硝胺诱发大鼠肝癌的的形成有促进作用,对免疫功能有明显抑制作用。
Objective:to observe The Effects of Jianpi Shugan Jiedu Recip on the Expression of p53 and survivin gene During DEN-Induced Hepatocarcinogenesis in Ra,and further explore its mechanism,with a view to effective clinical use of the JPSGJD recipe detoxification of liver cancer,especially cancer of advanced treatment provide an objective basis.
Methods:male healthy rat weighting 110±10g,(n=72).feeding 3 days in normal. Later,rat were randomly divided into 6 groups by SPSS:DEN Control group, Middle-dose of JPSGJD with DEN group,High dose of JPSGJD with DEN group; BINZHENGJIEH modle Control group of JPSGJD,Middle dose of JPSGJD group, High dose of JPSGJD group.the chemotherapy drug was given by intraperitoneal injection for all group to the end.DEN SolutionConcentration is 1mlDEN:10000ml pured Sterile water,the rat drank that Solution everyday,until the end of the experiment.To establishe the DEN-Induced Hepatocarcinogenesis in Rat model. Secondly,we give traditional Chinese medicine DAHUANGConcentrated liquid Ig for BINZHENGJIEH modle Control group's all rat,at the same time,Long-term constraint rat limbs,Restraint rat' food,activtation to anger them.Throught this way to establish PIXUGANYU model on Rat.the dosage of Middle dose of JPSGJD group and High dose of JPSGJD group isl0mg/kg,30mg/kg.At the end of the experiments,the rat were killed by cervical dislocation,we weighed the tumor,the spleen,the liver.Calculated the rate of inhibitory effect of liver carcinoma and the index of the spleen,the liver.HE stain,light microscopy level of metastasis.We measured P53、Survivin expression of the rat through Immunohistochemistry technique method.To explore the mechanism of the JPSGJD recipe on the liver cancer of mechanism.
Results:It has showed that the JPSGJD recipe has remarkably suppressing effects on cancer of the DEN-Induced Hepatocarcinogenesis in Rat.It can significantly increase the inhibition rate of DEN-Induced Hepatocarcinogenesis in rat of liver cancer(compared with the control group,P＜0.01);In addition,the JPSGJD recipe can significantly improve the mice's immunologic function,especially for High dose of the SGJPJD recipe(compared with the control group,P＜0.01).The JPSGJD recipe can decrease the expression of P53 and Survivin protein expression(compared with the control group,P＜0.01),and the JPSGJD recipe with DEN group which compared with the DEN group and model group could have statistic differences(P＜0.01).
Conclusion:The JPSGJD recipe can inhibit liver cancer in rat,Their role may be related to P53、Survivin gene reduce expression;The JPSGJD recipe can also enviously enhance the immunologic function of liver cancer rat.
方法:健康72只SD雄性大鼠,体重110士20g,正常饲养三天,称重后按SPSS随机分为DEN对照组,DEN中药高剂量组,DEN中药低剂量组;病证结合模型组,病证结合中药高剂量组,病证结合中药低剂量组。DEN水溶液按照浓度为1mlDEN:10000ml灭菌蒸馏水的比例配成,给予分组后各组大鼠自由饮用,每天饮用,连续22周,建立诱发性肝癌大鼠实验模型。病证结合模型各组大鼠采用先连续灌胃生大黄煎液,并慢性束缚大鼠四肢,限制活动和断食以施加脾虚肝郁刺激因子,同时给予DEN溶液自由饮用,建立脾虚肝郁大鼠肝癌病证模型。DEN中药高、低剂量组和病证结合中药高、低剂量组,分别按30mg/kg、10mg/kg给予中药,DEN对照组和病证模型组给予生理盐水,连续22周。实验结束后第2天,各组大鼠于乙醚麻醉下处死,观察肝脏表面病变结节,称取肝脏、脾脏重量,计算肝脏指数,脾指数。HE染色,光镜下观察大鼠肝脏组织结构,并运用免疫组化法检测肝脏组织突变型P53,Survivin的表达水平。
结果:大鼠经脾虚肝郁因素刺激和DEN作用下的病证结合模型组,与单纯DEN诱发性大鼠肝癌对照组比较,免疫器官的重量有显著下降,二者有显著性差异(P＜0.05);而健脾疏肝解毒方能显著提高诱癌大鼠免疫器官的重量,各组大鼠肝指数、脾指数结果与DEN对照组相比,DEN中药高、低剂量组有显著统计学意义(P＜0.01),中药高剂量组脾指数显著高于中药低剂量组(P＜0.01);中药高、低剂量组肝指数低于DEN对照组(P＜0.01);病证中药高、低剂量组脾指数均高于病证模型组,尤其是病证中药高剂量组更为显著(P＜0.01);病证中药高、低剂量组肝指数均低于病证模型组(P＜0.01)。健脾疏肝解毒方亦能降低肝癌组织突变型P53,Survivin蛋白的表达量,与DEN对照组相比,DEN中药高、低剂量组有明显差异(P＜0.01);与病证结合模型组相比,病证中药高、低剂量组均有统计学意义(P＜0.05),病证结合中药高剂量组差异显著(P＜0.01)。
结论:健脾疏肝解毒方能明显抑制大鼠肝癌的形成,并能增强肝癌大鼠免疫器官的重量,其作用机制可能与健脾疏肝解毒方下调肝癌组织突变P53,Survivin基因表达以及增强肝癌大鼠的免疫功能有关。并且脾虚肝郁刺激对二乙基亚硝胺诱发大鼠肝癌的的形成有促进作用,对免疫功能有明显抑制作用。
Objective:to observe The Effects of Jianpi Shugan Jiedu Recip on the Expression of p53 and survivin gene During DEN-Induced Hepatocarcinogenesis in Ra,and further explore its mechanism,with a view to effective clinical use of the JPSGJD recipe detoxification of liver cancer,especially cancer of advanced treatment provide an objective basis.
Methods:male healthy rat weighting 110±10g,(n=72).feeding 3 days in normal. Later,rat were randomly divided into 6 groups by SPSS:DEN Control group, Middle-dose of JPSGJD with DEN group,High dose of JPSGJD with DEN group; BINZHENGJIEH modle Control group of JPSGJD,Middle dose of JPSGJD group, High dose of JPSGJD group.the chemotherapy drug was given by intraperitoneal injection for all group to the end.DEN SolutionConcentration is 1mlDEN:10000ml pured Sterile water,the rat drank that Solution everyday,until the end of the experiment.To establishe the DEN-Induced Hepatocarcinogenesis in Rat model. Secondly,we give traditional Chinese medicine DAHUANGConcentrated liquid Ig for BINZHENGJIEH modle Control group's all rat,at the same time,Long-term constraint rat limbs,Restraint rat' food,activtation to anger them.Throught this way to establish PIXUGANYU model on Rat.the dosage of Middle dose of JPSGJD group and High dose of JPSGJD group isl0mg/kg,30mg/kg.At the end of the experiments,the rat were killed by cervical dislocation,we weighed the tumor,the spleen,the liver.Calculated the rate of inhibitory effect of liver carcinoma and the index of the spleen,the liver.HE stain,light microscopy level of metastasis.We measured P53、Survivin expression of the rat through Immunohistochemistry technique method.To explore the mechanism of the JPSGJD recipe on the liver cancer of mechanism.
Results:It has showed that the JPSGJD recipe has remarkably suppressing effects on cancer of the DEN-Induced Hepatocarcinogenesis in Rat.It can significantly increase the inhibition rate of DEN-Induced Hepatocarcinogenesis in rat of liver cancer(compared with the control group,P＜0.01);In addition,the JPSGJD recipe can significantly improve the mice's immunologic function,especially for High dose of the SGJPJD recipe(compared with the control group,P＜0.01).The JPSGJD recipe can decrease the expression of P53 and Survivin protein expression(compared with the control group,P＜0.01),and the JPSGJD recipe with DEN group which compared with the DEN group and model group could have statistic differences(P＜0.01).
Conclusion:The JPSGJD recipe can inhibit liver cancer in rat,Their role may be related to P53、Survivin gene reduce expression;The JPSGJD recipe can also enviously enhance the immunologic function of liver cancer rat.
引文
[1]Parkin D M.Global cancer statistics in the year 2000[J].Lancet Oncol,2001,2(9):533-543
[2]叶任高,陆再英.内科学[M].人民卫生出版社,2001,1:471
[3]叶家才,崔书中,巴明臣.原发性肝癌的流行病学特征及其危险因素[J].实用医学杂志,2008,24(10)
[4]叶丽红.抗癌扶正法治疗原发性肝癌的临床及机理研究[M].南京中医药大学博士学位论文,1999,9-13
[5]程学莲,王伯庆.浅谈脾虚与肝癌的相关性[J].吉林中医药,2007,8(27):8.
[6]王家晓.脾虚与肝癌的病因证治关系[J].中国中西医结合消化杂志,2005,2,1(1):63
[7]明·李梃.医学入门[M].南昌:江西科学技术出版社,1988:826
[8]于尔辛.肝癌的健脾理气方治疗[J].中国肿瘤,2000,9(8):344
[9]陈华圣,许爱华,王正兵,等.双苓扶正抗癌胶囊对小鼠肝癌的影响[J].中国实验方剂学杂志.2003,9(1):45
[10]王家晓,荣震,薛庭新.健脾益气中药对晚期肿瘤脾虚病人生活质量的影响[J].中医药临床杂志,2004,16(1):33
[11]韩克起,鲍利霞,黄传继.扶正抗癌方对大鼠移植性肝癌红细胞黏附功能及IL22水平的影响[J].中西医结合肝病杂志,2000,10(1):28
[12]吴万垠,钱耕荪,于尔辛.健脾理气合剂阻抑小鼠HBV协同AFBl致肝癌的机制[J].世界华人消化杂志,1998,6(7):594
[13]杨秉辉,夏景林,黄力文,等.我国肝癌“临床相”30年的变迁-原发性肝癌3250例的对比研究[J].中华医学杂志,2003,83
[14]崔岩.中医药在肝癌治疗中的应用体会[J].吉林中医药,2004,24(1):18-19
[15]蒋梅.健脾法对中晚期肝癌介入治疗后免疫功能的影响[J].江西中医学院学报,2000,12(3):101-124
[16]倪红梅.疏肝理气法在原发性肝癌中的治疗及预防中的应用[J].新中医2003,3,(3):3.777
[17]丛昌盛,于甬华.心理神经免疫学在肿瘤学研究中的进展[J].中华肿瘤防治杂 志.2008,15(8):635-637
[18]杨新中,李金彩,邹银水,张其军,谭晓云.对中医肿瘤病因病机与治疗的思考[J].中国医药学报,1999,14(6):57-60
[19]庞伴池,雷永仲.中医药治疗四例肝瘤的体会[J].中医杂志,1966,(5):33
[20]杨富春,郑树森,蒋天安.改良法大鼠原发性肝癌模型的建立.中华医学杂志,2004,1284(23):2018-2019
[21]徐淑云,卞如濂,陈修.药理实验方法学.北京:人民卫生出版社,2002,第三版:1427-1432
[22]马曾辰.可供临床研究的常用鼠肝癌模型[J].国外医学肿瘤分册,1982,1(1):13
[23]郑杰,武忠弼,阮幼冰,等.二乙基亚硝胺诱发大鼠肝癌过程中三种基因的原位表达[J].中华病理学杂志,1995,24(5):309-311
[24]OgawaK,YokokawaK,TomoyoriTetalInduction of gamma glutamyltranspeptidase positive altered hepatocytic lesions by combination of transplacental initiation and postanal selection [J].Int J Cancer,1982,29(3):333-336
[25]RabesHM,ScholzeP,JantschBG.rowthkineticsof diethylnitrosamineinduced,enzyme deifcient"preneoplastic"liver cell populations in vivo and in vitro[J]Cancer Res,1972,3,32(11):2577-2586
[26]Peraino C,Staffeldt EF,Carnes BA,et al Characte.rization of histo chemically detectable altered hepatocyte foci adn their relationship to hepatic tumorigenesis in rats treated once with diethylnitrosamine or benzo(a) pyrene within one day after birth[J].Cancer Res,1984,44(8):3340-3347
[27]Mitaka T,Tsukada H.Sexua difference in the histochemical characteristics of "altered cell foci"in the liver of aged Fischer 344 rats[J].Jpn J Cancer Res,1987,78(8):785-790
[28]王丹丹,张大方.诱发性大鼠肝癌模型的研究进展[J].吉林中医药,2006,10,26(10):75-76
[29]高进.肿瘤学基础与研究方法[M].北京人民卫生出版社,1999,1(1)
[30]陈志英,严瑞琪,覃国忠,等.建立黄曲霉毒素B1值致肝癌作用短期体内实验模型的进一步研究[J].广西医学院学报,1985,3(1):17
[31]段小娴,覃柳亮,覃国忠,等.单剂量黄曲霉毒素B1致大鼠肝癌作用的短期实验模型[J].癌症,1996,15(1):21-22
[32]齐秉均,李国利.二乙基亚硝胺和黄曲霉毒素B1联合诱发大鼠肝癌的研究[J].中华 病理学杂志,1990,19(3):184
[33]斯坎德尔·白克力,阿地里江·阿布力米提.诱发性肝癌动物模型在中药抗癌作用中的研究进展.[J]新疆医科大学学报,2008,10,31(10):1472
[34]葛锡锐,许河生,施渭康,等.二乙基亚硝胺(DENA)诱发大鼠肝癌过程甲胎蛋白变化的定性和定量观察[J].实验生物学报,1978,11(1):105-108
[35]Tessifore L,Tomasi C,Greo M,et al.A subnecrogenic dose of DENA is able to initiate hepatocarcin ogenesis in t he rat when coupled wit h fasting refeeding[J].Carcinogenesis,1996,17:289-292
[36]梁晓俐,钟汗光,左声鹤,等.二乙基亚硝胺诱发大鼠肝癌的病理形态学研究[J].第三军医大学学报,1991,13(1):32
[37]葛勇胜,许戈良.大鼠高转移肝癌模型的研究进展[J].国际外科杂志,2006,33(5):331-334
[38]岳利峰,丁杰,陈家旭,岳广欣,梁嫒,霍素坤,李晶晶.肝郁脾虚证大鼠模型的建立与评价[J].北京中医药大学学报,2008,6(31):396-400
[39]赵晓林,李恩,贾漪涛,等.滋补肝肾方药对慢性激怒应激大鼠下丘脑肝脏核糖体聚态的影响[J].中国中医基础医学杂志,1997,3(5):28-30
[40]谷俊朝,余微波,王宇,等.黄芪多糖对TA2小鼠乳腺癌MA2891移植瘤生长及HSP70表达的影响[J].中华肿瘤防治研究杂志,2006,13(20):1534-1537
[41]肖正明,赵联合,邱军,等.黄芪水提物对人肝癌细胞和瘤鼠免疫细胞的影响[J].山东中医药大学学报,2004,28(2):136-139
[42]杨雁,陈敏珠.黄芪总苷对肝癌细胞凋亡及wtp53基因表达的影响[J].中国药理学通报,2001,17(4):447-451
[43]申海涛,张祥宏,郑末,等.经口给予黄芪提取物对H22肝癌荷瘤小鼠影响的实验研究[J].中国肿瘤临床,2006,33(14):821-824
[44]党双锁,张正国,袁利超,等.大黄素和黄芪多糖对大鼠实验性肝癌的抑制作用[J].西安交通大学学报(医学版),2006,27(3):250-253
[45]杨雁,陈敏珠1黄芪总苷对肝癌细胞凋亡及wtp 53基因表达的影响1中国药理学通报,2001,17(4):447-451
[46]杨雁,宋少刚,陈敏珠.黄芪总提物对肝癌细胞生长、甲胎蛋白分泌及72GT活性的影响[J].中国临床药理学与治疗学,2001,6(1):182-201
[47]申志强,孙俊秀,黄立新.黄芪治疗选择性IgA缺乏的疗效[J].新药与临床,1997,16(4):246-247
[48]张艳,梁华平.黄芪多糖对烧伤小鼠细胞免疫功能的作用及机理探讨[J].中国实验临床免疫学杂志,1995,7(2):41
[49]张英,李铁兵,孙茂波.参芪合剂抗疲劳作用的基础与临床研究[J].中医药学报,1998(4):35
[50]侯世荣.黄芪口服液治疗慢性肝炎的疗效观察[J].中草药,2000,31(10):766
[51]马宏秀,张治祥.黄芪的药理研究进展[J].陕西中医学院学报,2004,9(5):27
[52]张宗,张鸿翔,史天良,等.白术挥发油的抗肿瘤作用的研究[J].肿瘤研究与临床,2006,18(12):799-820
[53]孙喜才,张玉五,连文太等.健脾益气方841-A对小鼠S180实体瘤增殖的影响[J].陕西中医,987,8(2):90-91
[54]关晓辉,曲娴,杨志萍等.白术挥发油对小鼠免疫功能的影响[J].北华大学学报(自然科学版),2001,2(2):122-123
[55]毛俊浩.白术多糖对小鼠淋巴细胞功能的调节[J].免疫学杂志,1996,12(4):233-236
[56]宿廷敏,王敏娟,阮时宝.白术的化学成分及药理作用研究概述[J].贵阳学院学报(自然科学版),2008,6(2):32-35
[57]刘仁义,张万峰,刘成德.温郁金1号注射液对急性中毒性肝炎疗效的病理形态学研究.中医药学报,1990(1):44
[58]贾宽.郁金挥发油对小鼠中毒性肝炎模型免疫功能的影响.中国免疫学杂志,1987,5(2):12
[59]李文喆,王勇,姚世芳.丹参药用成分研究进展[J].人民军医,2008,580:180-182
[60]敬静,王修杰,郑鸿等.丹参酮ⅡA对人ER阴性乳腺癌细胞的生长抑制和多耐药逆转作用[J].四川大学学报(医学版),2007,38(3):391-395
[61]张洪泉,柳丽.丹参活性成分的现代中药药理研究进展[J].中国野生植物资源,2003,22(6):12-41
[62]颜平,罗心平,施海明,等1丹参多酚酸治疗心绞痛患者的临床疗效及对血小板功能的影响[J]1介入放射学杂志,2004,12(2):55-59
[63]吴百灵.丹参对CCl4所致急性肝损伤保护作用的研究[J].中医药学刊,2002,20(3): ?361-381
[64]胡凯文,侯丽,陈信义等.大黄酸、大黄素抗多药耐药肿瘤细胞研究[J].中国中医基础药学杂志,1998,4(11):19
[65]洪照友,高毅,詹兴海.中药虎杖对大鼠肝脏缺血性损伤保护的形态学观察[J].世界华人消化杂志,2000,8(2):162-164
[66]刘志红,黎磊石,胡伟新.大黄素对肾小球系膜细胞c-myc原癌基因表达的影响[J].中国药理学报,1996,17(1):61
[67]姜晓峰,永苏.大黄素逆转肿瘤细胞多药抗药性作用[J].药学学报,1999,34(3):164
[68]Raymond F.Schinazi,Chuay K.Chu,J.Kamesh Babu,et al.Anthra guinones as a new class of antiviral agents against human immuno deficiency virus[J].Antiviral Research,1990,(13):265
[69]何球藻,吴厚生.医学免疫学.上海:上海医科大学出版社,1997,第二版:7-59
[70]吴咸中,高金亮.脾虚证的现代研究[M].天津科技译证出版社,1992:1-2:255-256
[71]彭桂英,顾立刚,王庆国,王济,李瑞生,王秀茜.肝郁脾虚因素刺激对DEN诱发肝癌大鼠免疫功能的影响[J].北京中医药大学学报,2002,11,25(6)41
[72]张海,刘秀芳,赵增虎,丁瑞亮,王炳胜,李成云.疏肝健脾法对肝癌肝动脉化疗栓塞后免疫功能的影响[J].临床军医杂志,2007,12,35(6):857-859
[73]XU Y,Regulation of p53 responses bypost-translationalmodifications[J].Cell Death Differ,2003,10(4):400-403
[74]SAGAR SENGUPTA,CURTIS C.Harris.P53:Traffic cop at the crossroads of repair and recombination[J].Nat Rev Mol Cell Biol,2005,6(1):44-55
[75]ATTARDI LD.The role of p53- mediated apoptosis as a crucial anti- tumor response to genomic instability:lessons from mouse models[J].Mutat Res,2005,569(1-2):145-157
[76]Hsu IC,Metcaff RA,Sun T,et al.Mutation hotspot in the p53 gene in human hepatocellular carcinona[J].Nature,1991,350(2):427
[77]Breasac B,Kew M,Wands J,et al.Abnormal structure and expression of p53 gene in human hepatocellular carcinoma[J].Proc Natl Acad Sci USA,1990,87(5):1973
[78]Chang SC,Lin JK,Lin TC,et al.Genetic alteration of p53,but not over expression of intratumoral p53 p rotein,or serum p53 antibody is a p rognostic factor in sporadic colorectal adenocarcinoma[J].nt JOncol,2005,26(1):65-75
[79]BergqvistM,Brattstrom D,Larsson A,et al.The role of circulating anti - p53 antibodies in patientswith advanced non - small cell lung cancer and their correlation to clinical parameters and survival[J].BMC Cancer,200414(4):66
[80]卢勤,滕皋军.p53基因治疗肿瘤的研究进展[J].中华放射学杂志,2000,34(10):659-661
[81]]任勇,陈寿松,李擒龙,等.肝细胞肝癌VEGF与p53的表达及其意义[J].华南国防医学杂志,2006,20(6):25-26
[82]Li FZ.Survivin study:what is the next wave?[J].Cell Physiol,2003,197(1):8-29
[83]Ambrosini G,Adida C,Altieri DC.A novel anti-apoptosisgene survivin expressed in cancer and lymphoma[J].NatMed,1997,3(8):917-921
[84]Ito T,Shiraki K,Sugimoto K,et al.Hepatocellularcareinoma associated wit h adult type cit rullinemia[J].DigDis Sci,2000,45(11):2203-2206
[85]Crossman D,Mc Niff JM,Li F.Expression of the apoptosis inhibitor,survivin,in nonmelanoma skin cancer and gene targeting in a keratinoyte cell line[J].Lab Invest,1999,79(9):1121-1126
[86]Suzuki A,Ito T,Kawano H,et al.Survivin initiates proeaspase p21 complex formation as a result of interaction with CDK4 to resist Fas-mediated cell death[J].Oncogene,2000,19(10):1346-1353
[87]Uren AG,Wang L,Pakusch M,et al.Survivin and the innercentromere protein INCENP show similar cell-cycle loealizationand gene knockout phenotype[J].Curt Biol,2000,10(21):1319-1328
[88]Muchmore SW,Chen J,Jakob C,et al.Crystal structure and mutagenic analysis of the inhibitor of apoptosis protein survivin[J].Mol Cell,2000,6(1):173-182
[89]计柳玉.Survivin在肝癌中的研究与应用[J].中国肿瘤,2006,15(3):180-182
[90]Elsayed YA,Sausville EA.Selected novel anticancer treatments targeting cell signaling proteins[J].Oncologist,2001,6(6):517-537
[91]王颖,王家.Survivin在原发性肝细胞癌中的表达及意义[J].中华消化杂志,2002,2(8):463-466
[92]Altieri DC.Validating survivin as a cancer therapeutic target[J].Nat Rev Cancer,2003,3(1):46-54
[93]Hoffman WH,Biade S,Zilfou JT,et al.Transcriptional repressionof the anti-apoptosic survivin gene by wild typep21[J].Biol Chem,2002,277(5):3247-3257
[94]陈明,伊新,林木生,缪辉来,陈念平,包仕廷.Survivin在肝癌中的表达及与p53表达的关系[J].中华肝胆外科杂志,2004,5(10):233
[95]O'Connor DS,Schechner S,Adida C,et al.Control ofapoptosis during angiogenesis by survivin expression inendothelial cells[J].Am J Pathol,2000,156(2):393-398
[96]Lu CD,AltieriDC,Tanigawa N,et al.Exp ression of a novel antiapop to sis gene,surviving,correlated with tumor cell apop tosis and p53 accumulation in gastric carcinomas [J].Cancer Res,1998,58(9):1808-1812
[97]曾建平,李衷,易继林.Survivin在肝癌中的表达及其与PCNA和P53表达的关系[J].Journal of Chinese Physician,2006,10(1 8):14-18。
[2]叶任高,陆再英.内科学[M].人民卫生出版社,2001,1:471
[3]叶家才,崔书中,巴明臣.原发性肝癌的流行病学特征及其危险因素[J].实用医学杂志,2008,24(10)
[4]叶丽红.抗癌扶正法治疗原发性肝癌的临床及机理研究[M].南京中医药大学博士学位论文,1999,9-13
[5]程学莲,王伯庆.浅谈脾虚与肝癌的相关性[J].吉林中医药,2007,8(27):8.
[6]王家晓.脾虚与肝癌的病因证治关系[J].中国中西医结合消化杂志,2005,2,1(1):63
[7]明·李梃.医学入门[M].南昌:江西科学技术出版社,1988:826
[8]于尔辛.肝癌的健脾理气方治疗[J].中国肿瘤,2000,9(8):344
[9]陈华圣,许爱华,王正兵,等.双苓扶正抗癌胶囊对小鼠肝癌的影响[J].中国实验方剂学杂志.2003,9(1):45
[10]王家晓,荣震,薛庭新.健脾益气中药对晚期肿瘤脾虚病人生活质量的影响[J].中医药临床杂志,2004,16(1):33
[11]韩克起,鲍利霞,黄传继.扶正抗癌方对大鼠移植性肝癌红细胞黏附功能及IL22水平的影响[J].中西医结合肝病杂志,2000,10(1):28
[12]吴万垠,钱耕荪,于尔辛.健脾理气合剂阻抑小鼠HBV协同AFBl致肝癌的机制[J].世界华人消化杂志,1998,6(7):594
[13]杨秉辉,夏景林,黄力文,等.我国肝癌“临床相”30年的变迁-原发性肝癌3250例的对比研究[J].中华医学杂志,2003,83
[14]崔岩.中医药在肝癌治疗中的应用体会[J].吉林中医药,2004,24(1):18-19
[15]蒋梅.健脾法对中晚期肝癌介入治疗后免疫功能的影响[J].江西中医学院学报,2000,12(3):101-124
[16]倪红梅.疏肝理气法在原发性肝癌中的治疗及预防中的应用[J].新中医2003,3,(3):3.777
[17]丛昌盛,于甬华.心理神经免疫学在肿瘤学研究中的进展[J].中华肿瘤防治杂 志.2008,15(8):635-637
[18]杨新中,李金彩,邹银水,张其军,谭晓云.对中医肿瘤病因病机与治疗的思考[J].中国医药学报,1999,14(6):57-60
[19]庞伴池,雷永仲.中医药治疗四例肝瘤的体会[J].中医杂志,1966,(5):33
[20]杨富春,郑树森,蒋天安.改良法大鼠原发性肝癌模型的建立.中华医学杂志,2004,1284(23):2018-2019
[21]徐淑云,卞如濂,陈修.药理实验方法学.北京:人民卫生出版社,2002,第三版:1427-1432
[22]马曾辰.可供临床研究的常用鼠肝癌模型[J].国外医学肿瘤分册,1982,1(1):13
[23]郑杰,武忠弼,阮幼冰,等.二乙基亚硝胺诱发大鼠肝癌过程中三种基因的原位表达[J].中华病理学杂志,1995,24(5):309-311
[24]OgawaK,YokokawaK,TomoyoriTetalInduction of gamma glutamyltranspeptidase positive altered hepatocytic lesions by combination of transplacental initiation and postanal selection [J].Int J Cancer,1982,29(3):333-336
[25]RabesHM,ScholzeP,JantschBG.rowthkineticsof diethylnitrosamineinduced,enzyme deifcient"preneoplastic"liver cell populations in vivo and in vitro[J]Cancer Res,1972,3,32(11):2577-2586
[26]Peraino C,Staffeldt EF,Carnes BA,et al Characte.rization of histo chemically detectable altered hepatocyte foci adn their relationship to hepatic tumorigenesis in rats treated once with diethylnitrosamine or benzo(a) pyrene within one day after birth[J].Cancer Res,1984,44(8):3340-3347
[27]Mitaka T,Tsukada H.Sexua difference in the histochemical characteristics of "altered cell foci"in the liver of aged Fischer 344 rats[J].Jpn J Cancer Res,1987,78(8):785-790
[28]王丹丹,张大方.诱发性大鼠肝癌模型的研究进展[J].吉林中医药,2006,10,26(10):75-76
[29]高进.肿瘤学基础与研究方法[M].北京人民卫生出版社,1999,1(1)
[30]陈志英,严瑞琪,覃国忠,等.建立黄曲霉毒素B1值致肝癌作用短期体内实验模型的进一步研究[J].广西医学院学报,1985,3(1):17
[31]段小娴,覃柳亮,覃国忠,等.单剂量黄曲霉毒素B1致大鼠肝癌作用的短期实验模型[J].癌症,1996,15(1):21-22
[32]齐秉均,李国利.二乙基亚硝胺和黄曲霉毒素B1联合诱发大鼠肝癌的研究[J].中华 病理学杂志,1990,19(3):184
[33]斯坎德尔·白克力,阿地里江·阿布力米提.诱发性肝癌动物模型在中药抗癌作用中的研究进展.[J]新疆医科大学学报,2008,10,31(10):1472
[34]葛锡锐,许河生,施渭康,等.二乙基亚硝胺(DENA)诱发大鼠肝癌过程甲胎蛋白变化的定性和定量观察[J].实验生物学报,1978,11(1):105-108
[35]Tessifore L,Tomasi C,Greo M,et al.A subnecrogenic dose of DENA is able to initiate hepatocarcin ogenesis in t he rat when coupled wit h fasting refeeding[J].Carcinogenesis,1996,17:289-292
[36]梁晓俐,钟汗光,左声鹤,等.二乙基亚硝胺诱发大鼠肝癌的病理形态学研究[J].第三军医大学学报,1991,13(1):32
[37]葛勇胜,许戈良.大鼠高转移肝癌模型的研究进展[J].国际外科杂志,2006,33(5):331-334
[38]岳利峰,丁杰,陈家旭,岳广欣,梁嫒,霍素坤,李晶晶.肝郁脾虚证大鼠模型的建立与评价[J].北京中医药大学学报,2008,6(31):396-400
[39]赵晓林,李恩,贾漪涛,等.滋补肝肾方药对慢性激怒应激大鼠下丘脑肝脏核糖体聚态的影响[J].中国中医基础医学杂志,1997,3(5):28-30
[40]谷俊朝,余微波,王宇,等.黄芪多糖对TA2小鼠乳腺癌MA2891移植瘤生长及HSP70表达的影响[J].中华肿瘤防治研究杂志,2006,13(20):1534-1537
[41]肖正明,赵联合,邱军,等.黄芪水提物对人肝癌细胞和瘤鼠免疫细胞的影响[J].山东中医药大学学报,2004,28(2):136-139
[42]杨雁,陈敏珠.黄芪总苷对肝癌细胞凋亡及wtp53基因表达的影响[J].中国药理学通报,2001,17(4):447-451
[43]申海涛,张祥宏,郑末,等.经口给予黄芪提取物对H22肝癌荷瘤小鼠影响的实验研究[J].中国肿瘤临床,2006,33(14):821-824
[44]党双锁,张正国,袁利超,等.大黄素和黄芪多糖对大鼠实验性肝癌的抑制作用[J].西安交通大学学报(医学版),2006,27(3):250-253
[45]杨雁,陈敏珠1黄芪总苷对肝癌细胞凋亡及wtp 53基因表达的影响1中国药理学通报,2001,17(4):447-451
[46]杨雁,宋少刚,陈敏珠.黄芪总提物对肝癌细胞生长、甲胎蛋白分泌及72GT活性的影响[J].中国临床药理学与治疗学,2001,6(1):182-201
[47]申志强,孙俊秀,黄立新.黄芪治疗选择性IgA缺乏的疗效[J].新药与临床,1997,16(4):246-247
[48]张艳,梁华平.黄芪多糖对烧伤小鼠细胞免疫功能的作用及机理探讨[J].中国实验临床免疫学杂志,1995,7(2):41
[49]张英,李铁兵,孙茂波.参芪合剂抗疲劳作用的基础与临床研究[J].中医药学报,1998(4):35
[50]侯世荣.黄芪口服液治疗慢性肝炎的疗效观察[J].中草药,2000,31(10):766
[51]马宏秀,张治祥.黄芪的药理研究进展[J].陕西中医学院学报,2004,9(5):27
[52]张宗,张鸿翔,史天良,等.白术挥发油的抗肿瘤作用的研究[J].肿瘤研究与临床,2006,18(12):799-820
[53]孙喜才,张玉五,连文太等.健脾益气方841-A对小鼠S180实体瘤增殖的影响[J].陕西中医,987,8(2):90-91
[54]关晓辉,曲娴,杨志萍等.白术挥发油对小鼠免疫功能的影响[J].北华大学学报(自然科学版),2001,2(2):122-123
[55]毛俊浩.白术多糖对小鼠淋巴细胞功能的调节[J].免疫学杂志,1996,12(4):233-236
[56]宿廷敏,王敏娟,阮时宝.白术的化学成分及药理作用研究概述[J].贵阳学院学报(自然科学版),2008,6(2):32-35
[57]刘仁义,张万峰,刘成德.温郁金1号注射液对急性中毒性肝炎疗效的病理形态学研究.中医药学报,1990(1):44
[58]贾宽.郁金挥发油对小鼠中毒性肝炎模型免疫功能的影响.中国免疫学杂志,1987,5(2):12
[59]李文喆,王勇,姚世芳.丹参药用成分研究进展[J].人民军医,2008,580:180-182
[60]敬静,王修杰,郑鸿等.丹参酮ⅡA对人ER阴性乳腺癌细胞的生长抑制和多耐药逆转作用[J].四川大学学报(医学版),2007,38(3):391-395
[61]张洪泉,柳丽.丹参活性成分的现代中药药理研究进展[J].中国野生植物资源,2003,22(6):12-41
[62]颜平,罗心平,施海明,等1丹参多酚酸治疗心绞痛患者的临床疗效及对血小板功能的影响[J]1介入放射学杂志,2004,12(2):55-59
[63]吴百灵.丹参对CCl4所致急性肝损伤保护作用的研究[J].中医药学刊,2002,20(3): ?361-381
[64]胡凯文,侯丽,陈信义等.大黄酸、大黄素抗多药耐药肿瘤细胞研究[J].中国中医基础药学杂志,1998,4(11):19
[65]洪照友,高毅,詹兴海.中药虎杖对大鼠肝脏缺血性损伤保护的形态学观察[J].世界华人消化杂志,2000,8(2):162-164
[66]刘志红,黎磊石,胡伟新.大黄素对肾小球系膜细胞c-myc原癌基因表达的影响[J].中国药理学报,1996,17(1):61
[67]姜晓峰,永苏.大黄素逆转肿瘤细胞多药抗药性作用[J].药学学报,1999,34(3):164
[68]Raymond F.Schinazi,Chuay K.Chu,J.Kamesh Babu,et al.Anthra guinones as a new class of antiviral agents against human immuno deficiency virus[J].Antiviral Research,1990,(13):265
[69]何球藻,吴厚生.医学免疫学.上海:上海医科大学出版社,1997,第二版:7-59
[70]吴咸中,高金亮.脾虚证的现代研究[M].天津科技译证出版社,1992:1-2:255-256
[71]彭桂英,顾立刚,王庆国,王济,李瑞生,王秀茜.肝郁脾虚因素刺激对DEN诱发肝癌大鼠免疫功能的影响[J].北京中医药大学学报,2002,11,25(6)41
[72]张海,刘秀芳,赵增虎,丁瑞亮,王炳胜,李成云.疏肝健脾法对肝癌肝动脉化疗栓塞后免疫功能的影响[J].临床军医杂志,2007,12,35(6):857-859
[73]XU Y,Regulation of p53 responses bypost-translationalmodifications[J].Cell Death Differ,2003,10(4):400-403
[74]SAGAR SENGUPTA,CURTIS C.Harris.P53:Traffic cop at the crossroads of repair and recombination[J].Nat Rev Mol Cell Biol,2005,6(1):44-55
[75]ATTARDI LD.The role of p53- mediated apoptosis as a crucial anti- tumor response to genomic instability:lessons from mouse models[J].Mutat Res,2005,569(1-2):145-157
[76]Hsu IC,Metcaff RA,Sun T,et al.Mutation hotspot in the p53 gene in human hepatocellular carcinona[J].Nature,1991,350(2):427
[77]Breasac B,Kew M,Wands J,et al.Abnormal structure and expression of p53 gene in human hepatocellular carcinoma[J].Proc Natl Acad Sci USA,1990,87(5):1973
[78]Chang SC,Lin JK,Lin TC,et al.Genetic alteration of p53,but not over expression of intratumoral p53 p rotein,or serum p53 antibody is a p rognostic factor in sporadic colorectal adenocarcinoma[J].nt JOncol,2005,26(1):65-75
[79]BergqvistM,Brattstrom D,Larsson A,et al.The role of circulating anti - p53 antibodies in patientswith advanced non - small cell lung cancer and their correlation to clinical parameters and survival[J].BMC Cancer,200414(4):66
[80]卢勤,滕皋军.p53基因治疗肿瘤的研究进展[J].中华放射学杂志,2000,34(10):659-661
[81]]任勇,陈寿松,李擒龙,等.肝细胞肝癌VEGF与p53的表达及其意义[J].华南国防医学杂志,2006,20(6):25-26
[82]Li FZ.Survivin study:what is the next wave?[J].Cell Physiol,2003,197(1):8-29
[83]Ambrosini G,Adida C,Altieri DC.A novel anti-apoptosisgene survivin expressed in cancer and lymphoma[J].NatMed,1997,3(8):917-921
[84]Ito T,Shiraki K,Sugimoto K,et al.Hepatocellularcareinoma associated wit h adult type cit rullinemia[J].DigDis Sci,2000,45(11):2203-2206
[85]Crossman D,Mc Niff JM,Li F.Expression of the apoptosis inhibitor,survivin,in nonmelanoma skin cancer and gene targeting in a keratinoyte cell line[J].Lab Invest,1999,79(9):1121-1126
[86]Suzuki A,Ito T,Kawano H,et al.Survivin initiates proeaspase p21 complex formation as a result of interaction with CDK4 to resist Fas-mediated cell death[J].Oncogene,2000,19(10):1346-1353
[87]Uren AG,Wang L,Pakusch M,et al.Survivin and the innercentromere protein INCENP show similar cell-cycle loealizationand gene knockout phenotype[J].Curt Biol,2000,10(21):1319-1328
[88]Muchmore SW,Chen J,Jakob C,et al.Crystal structure and mutagenic analysis of the inhibitor of apoptosis protein survivin[J].Mol Cell,2000,6(1):173-182
[89]计柳玉.Survivin在肝癌中的研究与应用[J].中国肿瘤,2006,15(3):180-182
[90]Elsayed YA,Sausville EA.Selected novel anticancer treatments targeting cell signaling proteins[J].Oncologist,2001,6(6):517-537
[91]王颖,王家.Survivin在原发性肝细胞癌中的表达及意义[J].中华消化杂志,2002,2(8):463-466
[92]Altieri DC.Validating survivin as a cancer therapeutic target[J].Nat Rev Cancer,2003,3(1):46-54
[93]Hoffman WH,Biade S,Zilfou JT,et al.Transcriptional repressionof the anti-apoptosic survivin gene by wild typep21[J].Biol Chem,2002,277(5):3247-3257
[94]陈明,伊新,林木生,缪辉来,陈念平,包仕廷.Survivin在肝癌中的表达及与p53表达的关系[J].中华肝胆外科杂志,2004,5(10):233
[95]O'Connor DS,Schechner S,Adida C,et al.Control ofapoptosis during angiogenesis by survivin expression inendothelial cells[J].Am J Pathol,2000,156(2):393-398
[96]Lu CD,AltieriDC,Tanigawa N,et al.Exp ression of a novel antiapop to sis gene,surviving,correlated with tumor cell apop tosis and p53 accumulation in gastric carcinomas [J].Cancer Res,1998,58(9):1808-1812
[97]曾建平,李衷,易继林.Survivin在肝癌中的表达及其与PCNA和P53表达的关系[J].Journal of Chinese Physician,2006,10(1 8):14-18。