皂荚提取物对小鼠肝癌细胞TGF-β_1信号传导通路的影响
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摘要
目的：据近年来研究发现：转化生长因子-β(TGF-β)作为一个多功能的生长因子,与肝癌等多种肿瘤的发生发展有着密切的关系。TGF-β信号通路的异常是肝癌发生发展及浸润和转移的重要因素之一.中医在治疗肝癌方面有其特色性,其中中药皂荚常作为治疗肿瘤的常用药物之一,近年来研究进展证实,中药皂荚提取物浓缩液具有抗癌的特性,可以有效抑制多种人类固体肿瘤,包括抑制肝癌细胞的生长。本实验主要探讨皂荚提取物对小鼠肝癌细胞TGF-β1信号通路系统调节的影响,为临床中药抗肿瘤治疗提供实验依据。
方法：本研究取清洁级昆明种小鼠60只,除空白对照组小鼠10只外,其余50只小鼠均予肝癌细胞株H22移植,建立小鼠右上肢皮下移植肿瘤模型,成瘤后50只小鼠被随机分为5组进行灌胃处理,分别为：皂荚提取物组(高剂量组：18.02g/次；中剂量组：9.01g/次；低剂量组：4.505g/次)、金龙胶囊组(阳性对照组：9.01g)、生理盐水组(模型组：9.01ml)上述5组均按剂量给药,每日三次、另空白对照组10只(正常喂养,不做任何处理)。用专用小鼠灌胃针持续灌胃给药15天以后,于第16天处死所有小白鼠,并采取样本(肝脏组织,眼球取血等),进行肝癌细胞生化学(肝功能,a-L-岩藻糖苷酶(AFU))及分子生物学(TGF-β1表达)等指标检测。
结果：根据实验数据结果进行分析：给药后,皂荚提取物高、中、低各组小鼠与金龙胶囊组小鼠TGF-β1表达阳性率明显高于模型组和空白对照组；皂荚提取物组小鼠AFU和肝功能检测指标降低,与模型组小鼠进行统计学分析比较,有统计学意义(P<0.05)。
结论：皂荚提取物能降低肝癌小鼠AFU的水平；减轻对肝癌细胞对小鼠肝脏的损害；增强TGF-β1的表达,达到抑制小鼠肝癌细胞的增殖,并促进肝癌细胞的凋亡。
Objective: According to recent study found that: transforming growth factor-β(TGF-β) as a multifunctional growth factor, and liver cancer and othertumor development and are closely related. TGF-βsignaling pathway is abnormal liver cancer invasion and metastasis development and an important factor. Chinese medicine in the treatment of liver cancer has its own characteristics, which traditional Chinese medicine as a treatment for tumors of Gleditsia often one of the commonly used drugs, recent research confirmed that the Chinese Gleditsia extract concentrate with anti-cancer properties, can inhibit a variety of human solid tumors, including liver cancer cell growth inhibition. This study, we investigated extract on liver cancer cells Gleditsia TGF-β1 signaling system regulating effects of anti-tumor therapy in clinical medicine to provide experimental basis.
Methods: Kunming mice of clean grade were obtained 60, in addition to 10 control mice, the remaining 50 mice were correspondingly transplanted hepatoma cell line H22, the establishment of the right upper limb subcutaneous mouse tumor model, tumor hfter the 50 mice were randomly divided into 5 groups fed treatments were: Gleditsia extract group (high dose group: 18.02g / times; middie dose group: 9.01g / times; low dose group: 4.505g / times ), Jinlong Capsule group (positive control group: 9.01g), saline group (model group: 9.01ml) above 5 groups all give three times a day, and the other 10 control group (normal feeding, not any treatment), After 15 days of continuous oral administration, in the first 16 days all mice were sacrificed, and to take samples (the liver, eyes, blood, etc.), to liver cell biochemistry [liver function, a-L-fucosidase (AFU)], and Molecular Biology (TGF-β1 expression) and so on detection.
Results: According to the results of analysis of experimental data, after the administration, Gleditsia extract high, medium and low groups Jinlong Capsule group mice and TGF-β1 expression in mice was significantly higher than model group and control group; Gleditsia extract group AFU and liver function tests in mice decreased with the model index for statistical analysis and comparison of mice, there was significant (P<0.05).
Conclusion: Gleditsia extract on liver cancer cells TGF-β1 signaling system to regulate, can enhance the expression of TGF-β1 to inhibit the proliferation of hepatoma ceils and promote apoptosis of hepatoma cells.
方法：本研究取清洁级昆明种小鼠60只,除空白对照组小鼠10只外,其余50只小鼠均予肝癌细胞株H22移植,建立小鼠右上肢皮下移植肿瘤模型,成瘤后50只小鼠被随机分为5组进行灌胃处理,分别为：皂荚提取物组(高剂量组：18.02g/次；中剂量组：9.01g/次；低剂量组：4.505g/次)、金龙胶囊组(阳性对照组：9.01g)、生理盐水组(模型组：9.01ml)上述5组均按剂量给药,每日三次、另空白对照组10只(正常喂养,不做任何处理)。用专用小鼠灌胃针持续灌胃给药15天以后,于第16天处死所有小白鼠,并采取样本(肝脏组织,眼球取血等),进行肝癌细胞生化学(肝功能,a-L-岩藻糖苷酶(AFU))及分子生物学(TGF-β1表达)等指标检测。
结果：根据实验数据结果进行分析：给药后,皂荚提取物高、中、低各组小鼠与金龙胶囊组小鼠TGF-β1表达阳性率明显高于模型组和空白对照组；皂荚提取物组小鼠AFU和肝功能检测指标降低,与模型组小鼠进行统计学分析比较,有统计学意义(P<0.05)。
结论：皂荚提取物能降低肝癌小鼠AFU的水平；减轻对肝癌细胞对小鼠肝脏的损害；增强TGF-β1的表达,达到抑制小鼠肝癌细胞的增殖,并促进肝癌细胞的凋亡。
Objective: According to recent study found that: transforming growth factor-β(TGF-β) as a multifunctional growth factor, and liver cancer and othertumor development and are closely related. TGF-βsignaling pathway is abnormal liver cancer invasion and metastasis development and an important factor. Chinese medicine in the treatment of liver cancer has its own characteristics, which traditional Chinese medicine as a treatment for tumors of Gleditsia often one of the commonly used drugs, recent research confirmed that the Chinese Gleditsia extract concentrate with anti-cancer properties, can inhibit a variety of human solid tumors, including liver cancer cell growth inhibition. This study, we investigated extract on liver cancer cells Gleditsia TGF-β1 signaling system regulating effects of anti-tumor therapy in clinical medicine to provide experimental basis.
Methods: Kunming mice of clean grade were obtained 60, in addition to 10 control mice, the remaining 50 mice were correspondingly transplanted hepatoma cell line H22, the establishment of the right upper limb subcutaneous mouse tumor model, tumor hfter the 50 mice were randomly divided into 5 groups fed treatments were: Gleditsia extract group (high dose group: 18.02g / times; middie dose group: 9.01g / times; low dose group: 4.505g / times ), Jinlong Capsule group (positive control group: 9.01g), saline group (model group: 9.01ml) above 5 groups all give three times a day, and the other 10 control group (normal feeding, not any treatment), After 15 days of continuous oral administration, in the first 16 days all mice were sacrificed, and to take samples (the liver, eyes, blood, etc.), to liver cell biochemistry [liver function, a-L-fucosidase (AFU)], and Molecular Biology (TGF-β1 expression) and so on detection.
Results: According to the results of analysis of experimental data, after the administration, Gleditsia extract high, medium and low groups Jinlong Capsule group mice and TGF-β1 expression in mice was significantly higher than model group and control group; Gleditsia extract group AFU and liver function tests in mice decreased with the model index for statistical analysis and comparison of mice, there was significant (P<0.05).
Conclusion: Gleditsia extract on liver cancer cells TGF-β1 signaling system to regulate, can enhance the expression of TGF-β1 to inhibit the proliferation of hepatoma ceils and promote apoptosis of hepatoma cells.
引文
[1]Sherman M. Hepatocellular carcinoma:epidemiology, risk factors, and screeing[J].Semin Liver Dis, 2005,25:143-154.
[2]Subramanian G, Schwarz R E, aiggins L, et al. Targeting endogenous transforming growth factor beta receptor signaling in SMAD4 deficient pancreatic carcinoma cells inhibits their invaaive phenotypel [J]. Cancer Res, 2004,64(15):5200-5211.
[3]Akiko Hatal. Molecular Cardiology Research Institute. TGF-β signaling and Cancer. Expreimental Cell Research 264,111-116(2001)
[4]Aristidis Moustakas,Serhiy Souchelnytskyi. Smad regulation in TGF-β signal tranaduction. Journal of Cell Science 114,4359-4369(2001)
[5]申景岭,张小玲.TGF-β/Smads信号转导途径与肿瘤发生发展的研究进展.国外医学遗传学分册[J],2003,26(3)：163-166.
[6]张振宇,张赤志,许汉林.等.皂荚提取物对人肝癌细胞相关基因表达的调控作用[J].中西医结合肝病杂志,2008,18(2)：93-95.
[7]香港理工大学.中药皂荚浓缩液具有抗癌特性[J].山东中医药大学学报,2005,31(4)：286.
[8]佘小湟,戴西湖.中药在原发性肝癌治疗中的作用.中西医结合肝病杂志,1998,8(1)：55.
[9]孔丽,姚树坤,马英,等.TGF-β1/信号转导通路的变化在原发性肝癌发病机制中的作用.临床肝胆病杂志,2007；23(4)：270-272
[10]李艳目.皂荚树的利用价值与栽培技术[J].现代农业科技,2008；(13)：85-86
[11]黄雯霞.肝内注射华蟾素治疗肝癌18例初步观察.癌症,1990；9(3)：239
[12]李茂,李伟芳,覃良,等.九节龙皂甙体内的抗肿瘤作用[J].广西中医学院学报,2004；7(2)：11-12.
[13]胡秀兰,胡柏林,韩世亮,等.武汉所昆明小鼠生产性能试验研究.中国实验动物学杂志,2002；4(1)：14-16.
[14]杨甲梅.肝脏占位性病变患者血清a-L-岩藻糖苷酶活性测定及其临床意义.中华医学检验杂志,1990；(1)
[15]钱伯文.治疗原发性肝癌经验.《中医杂志》,1999；(8)
[16]陈友芝.晚期肝癌的中药治疗.《浙江中医学院学报》,1990；(14)：1
[1]、谢怡庄,叶新苗.肝癌的中医治疗原则探讨.中国医药学报,2004；(19)： 3.
[2]、李智,杜杰.原发性肝癌的中医治疗研究进展.实用中医内科杂志,1997；(11)：1.
[3]、魏琳,杨晨光.中医辨证与辨病相结合治疗晚期原发性肝癌52例.陕西中医,2002；(23)
[4]、王文宝.肝癌的中医治疗呈现三大变化.现代医院报,2005；(2).
[5]、陈树森,李智.原发性肝癌中医治疗研究进展概述.实用中医内科杂志,1997；(11)：1.
[6]、李佐清.中药治疗原慢性肝癌的概况.中医杂志,1996；37(5)：308-310.
[7]、乔红梅,贺新怀,席孝贤.中药抗肿痛机制研究浅析.陕西中医,2003；24(5)：454-456.
[8]、张银娣,沈建平等.黄芪皂苷甲对血浆cAMP及再生肝DNA合成的影响.药学学报,1984；(8).
[9]、徐进.SMAD4研究的新进展.国外医学生理病理科学与临床分册,2004；22(3)：310-312.
[10]、申景岭,张小玲.TGF-β/Smads信号转导途径与肿瘤发生发展的研究进展.国外医学遗传学分册[J],2003,；26(3)：163-166.
[11]、刘政,季国忠,缪林,等.人肝癌及癌旁组织中Smad4、Smad7 mRNA 和蛋白表达研究[J].天津医药,2004；32(12)：721-723.
[12]、张振宇,张赤志,许汉林等.皂荚提取物对人肝癌细胞相关基因表达的调控作用[J].中西医结合肝病杂志,2008；18(2)：93-95.
[13]、胡玲,王洪琦等.白花蛇舌草诱导HSP-(70)表达对H22肝癌细胞移植瘤的干预作用.中药新药与临床药理,2006；(6)
[14]、吴万垠,于尔辛.中医药对肿痛阻断作用的研究进展.中医杂志,1998；39(5)：308-310.
[15]、黄金昶.原发性肝癌中医治疗体会.中国临床医生,2006；(34)：10.
[16]、钱伯文.治疗原发性肝癌经验.中医杂志,1999；(8)
[17]、陈友芝.晚期肝癌的中药治疗.浙江中医学院学报,1990；(14)：1.
[18]、马伯亭.60例原发性肝癌临床疗效观察.中医药学报,1992；(5)：21-24.
[19]、孙亚林,于连荣.齐刺刘针法治疗肝癌疼痛80例疗效观察.中国针灸,2000；20(4)：211-212.
[20]、李金昌.中医辨证配合介入治疗原发性肝癌的临床观察.广东医学学,2001；3(22)：3.
[21]、吕爱林,张岳正.中医辨证配合介入疗法治疗中晚期肝癌30例.陕西中医,2004；(25)：9.
[22]、陈乃杰,金源.中医辨证配合放射治疗原发性肝癌的临床观察.肿瘤,1998；7(18)：4.
[23]、蛇毒胶囊合中医辨证治疗原发性肝癌疗效观察.浙江中西医结合杂 志,2005；(15)：9.
[24]、张丽,张华.中医姑息治疗肝癌.现代中西医结合杂志,2007；(11).
[25]、何兴翌,韩文.肝癌化疗配合中医-扶正祛邪-疏肝解郁-治疗护理体会.内蒙古中医药,2006；2(46)：1.
[2]Subramanian G, Schwarz R E, aiggins L, et al. Targeting endogenous transforming growth factor beta receptor signaling in SMAD4 deficient pancreatic carcinoma cells inhibits their invaaive phenotypel [J]. Cancer Res, 2004,64(15):5200-5211.
[3]Akiko Hatal. Molecular Cardiology Research Institute. TGF-β signaling and Cancer. Expreimental Cell Research 264,111-116(2001)
[4]Aristidis Moustakas,Serhiy Souchelnytskyi. Smad regulation in TGF-β signal tranaduction. Journal of Cell Science 114,4359-4369(2001)
[5]申景岭,张小玲.TGF-β/Smads信号转导途径与肿瘤发生发展的研究进展.国外医学遗传学分册[J],2003,26(3)：163-166.
[6]张振宇,张赤志,许汉林.等.皂荚提取物对人肝癌细胞相关基因表达的调控作用[J].中西医结合肝病杂志,2008,18(2)：93-95.
[7]香港理工大学.中药皂荚浓缩液具有抗癌特性[J].山东中医药大学学报,2005,31(4)：286.
[8]佘小湟,戴西湖.中药在原发性肝癌治疗中的作用.中西医结合肝病杂志,1998,8(1)：55.
[9]孔丽,姚树坤,马英,等.TGF-β1/信号转导通路的变化在原发性肝癌发病机制中的作用.临床肝胆病杂志,2007；23(4)：270-272
[10]李艳目.皂荚树的利用价值与栽培技术[J].现代农业科技,2008；(13)：85-86
[11]黄雯霞.肝内注射华蟾素治疗肝癌18例初步观察.癌症,1990；9(3)：239
[12]李茂,李伟芳,覃良,等.九节龙皂甙体内的抗肿瘤作用[J].广西中医学院学报,2004；7(2)：11-12.
[13]胡秀兰,胡柏林,韩世亮,等.武汉所昆明小鼠生产性能试验研究.中国实验动物学杂志,2002；4(1)：14-16.
[14]杨甲梅.肝脏占位性病变患者血清a-L-岩藻糖苷酶活性测定及其临床意义.中华医学检验杂志,1990；(1)
[15]钱伯文.治疗原发性肝癌经验.《中医杂志》,1999；(8)
[16]陈友芝.晚期肝癌的中药治疗.《浙江中医学院学报》,1990；(14)：1
[1]、谢怡庄,叶新苗.肝癌的中医治疗原则探讨.中国医药学报,2004；(19)： 3.
[2]、李智,杜杰.原发性肝癌的中医治疗研究进展.实用中医内科杂志,1997；(11)：1.
[3]、魏琳,杨晨光.中医辨证与辨病相结合治疗晚期原发性肝癌52例.陕西中医,2002；(23)
[4]、王文宝.肝癌的中医治疗呈现三大变化.现代医院报,2005；(2).
[5]、陈树森,李智.原发性肝癌中医治疗研究进展概述.实用中医内科杂志,1997；(11)：1.
[6]、李佐清.中药治疗原慢性肝癌的概况.中医杂志,1996；37(5)：308-310.
[7]、乔红梅,贺新怀,席孝贤.中药抗肿痛机制研究浅析.陕西中医,2003；24(5)：454-456.
[8]、张银娣,沈建平等.黄芪皂苷甲对血浆cAMP及再生肝DNA合成的影响.药学学报,1984；(8).
[9]、徐进.SMAD4研究的新进展.国外医学生理病理科学与临床分册,2004；22(3)：310-312.
[10]、申景岭,张小玲.TGF-β/Smads信号转导途径与肿瘤发生发展的研究进展.国外医学遗传学分册[J],2003,；26(3)：163-166.
[11]、刘政,季国忠,缪林,等.人肝癌及癌旁组织中Smad4、Smad7 mRNA 和蛋白表达研究[J].天津医药,2004；32(12)：721-723.
[12]、张振宇,张赤志,许汉林等.皂荚提取物对人肝癌细胞相关基因表达的调控作用[J].中西医结合肝病杂志,2008；18(2)：93-95.
[13]、胡玲,王洪琦等.白花蛇舌草诱导HSP-(70)表达对H22肝癌细胞移植瘤的干预作用.中药新药与临床药理,2006；(6)
[14]、吴万垠,于尔辛.中医药对肿痛阻断作用的研究进展.中医杂志,1998；39(5)：308-310.
[15]、黄金昶.原发性肝癌中医治疗体会.中国临床医生,2006；(34)：10.
[16]、钱伯文.治疗原发性肝癌经验.中医杂志,1999；(8)
[17]、陈友芝.晚期肝癌的中药治疗.浙江中医学院学报,1990；(14)：1.
[18]、马伯亭.60例原发性肝癌临床疗效观察.中医药学报,1992；(5)：21-24.
[19]、孙亚林,于连荣.齐刺刘针法治疗肝癌疼痛80例疗效观察.中国针灸,2000；20(4)：211-212.
[20]、李金昌.中医辨证配合介入治疗原发性肝癌的临床观察.广东医学学,2001；3(22)：3.
[21]、吕爱林,张岳正.中医辨证配合介入疗法治疗中晚期肝癌30例.陕西中医,2004；(25)：9.
[22]、陈乃杰,金源.中医辨证配合放射治疗原发性肝癌的临床观察.肿瘤,1998；7(18)：4.
[23]、蛇毒胶囊合中医辨证治疗原发性肝癌疗效观察.浙江中西医结合杂 志,2005；(15)：9.
[24]、张丽,张华.中医姑息治疗肝癌.现代中西医结合杂志,2007；(11).
[25]、何兴翌,韩文.肝癌化疗配合中医-扶正祛邪-疏肝解郁-治疗护理体会.内蒙古中医药,2006；2(46)：1.