中医五脏相关理论探讨及补脾益肾法对大鼠MSCs增殖和分化的影响
|
摘要
研究目的:
对五脏相关理论进行梳理和探讨,在此基础上,对子系统“脾肾相关”理论进行更为深入的研究,提出“骨髓间充质干细胞(MSCs)为脾肾相关细胞层次表达的存在形式”的工作假说,探讨补脾益肾法对大鼠MSCs增殖和分化的影响,从微观角度和干细胞水平研究脾肾相关。
研究方法:
一、五脏相关理论探讨
(一)五脏相关学说的缘起:
1.五脏相关源流探讨:经考证,词义最接近、年代最早的文献是《素问·玉机真脏论》:“五脏相通,移皆有次”。古人通过阐述五脏功能互用和脏腑相合相通理论、气机升降出入、疾病的传变、五脏旁通之说、天人相应及药性归经理论体现五脏之气相通。
2.中医五脏相关学说的形成与发展:邓铁涛教授在几十年的临床实践中,把五行学说和脏腑学说结合起来,提出了“五脏相关”学说。
(1)理论研究继续深化:90年代末本世纪初,邓铁涛教授指导两位博士研究生赵氏、郑氏对五脏相关学说科学内涵进行探讨。认为五脏相关学说是对中医经典理论五行学说的继承和发展,具有跨学科的特点。
(2)五脏相关学说临床研究:五脏相关临床研究是目前研究的重点,主要有两个层次的研究,一是五脏相关总体层面的研究,二是深入到子系统如两脏相关的研究。面上的研究显示五脏相关理论已经广泛应用于临床各个系统疾病的治疗,与此同时,各个子系统的研究也逐渐开展。目前,以两两相关的研究较多。如脾(胃)肾相关,心脾(胃)相关,肝脾(胃)相关,胆胃相关,肺脾(胃)相关,心肺相关,心胆相关,心肾相关,肺肾相关,肝肾相关,肝肺相关等。
(3)五脏相关学说实验研究:部分学者进行了实验研究,揭示了五脏中的肝、脾(这里指空肠绒毛)、肾之间的相生、相克关系,“肺肾相关”的物质基础,中医“心肾相关”理论的科学内涵等。
(二)脾肾相关子系统研究:
中医脾肾相关子系统以阐发脾和肾相关的生理功能、病理变化机理及其证治方药为中心内容,体现了脏腑同病病机和整体观念。与传统的中医理论比较,脾肾相关吸取了五行学说中“土制水”的精神内核,并综合了历代医家对脾肾相互关系的不断发展,而将二者之间动态的、错综复杂的相互关系用“相关”来概括。近年来,国内有
Objective:This topic discussed and systematized the theory of "spleen-kidney related" on the basic of literature. Which was one of subsystem of "the doctrine of the correlation of five viscera" put forward by Professor Tietao Deng, the famous TCM scientist of the contemporary. We would induce the frequent type and medication rule of Flaccidity syndrome in clinical part. While in experimental part, we approach the effect on multiplication and differentiation of Mesenchymal stem cells (MSCs) resulting from "Qiangjijianli" fluid for oral use. On the other hand, we' d like to discuss the mechanism of action of the medicine in curing Flaccidity syndrome.Methods:1. Theory researchThe original literature of the doctrine of the correlation of five viscera is "plain questions" .The ancients explain the communication of QI of five ZANG-organs. The theory are put forward by Professor tietao deng on the basic of clinical practice, who integrate theory of five-element and zang-fu organs. The theory are developed by his students, while are widely studied on clinical application and empirical study. one of subsystem of "the doctrine of the correlation of five viscera" was the theory of spleen-kidney related.2. Empirical study: Isolation and culture of MSCsHeparinized bone marrow was obtained from SD rat. The marrow samples were diluted with Is cove' s modified Dovecot' s medium (IMDM) containing 5% fetal bovine serum (FBS) and 5% horse serum (HS). The mononuclear cells were prepared by gradient centrifugation at 2000rpm. The cells were washed, counted and
plated at 10000 cells/75cm2 on Petri dishes in IMDM supplemental with 5% FBS and 5% HS which was selected for proper recovery and expansion of MSCs. The cells were incubated in a humidified incubator in 5% co2 at 37°C. Medium was replaced and the cells in suspension were removed at 48 hours and every three days thereafter. MSCs formed as confluent layers were detached by treatment with 0.25% trypsin(sigma) and seeded into cultural flasks. MSCs after the third subculture were used for the described experiments. Analysis of cell surface moleculesThe fixed cells were harvested from tissue culture flasks. Aliquots were incubated for one night at refrigerator 4°C with Multiclone antibody against rabbit CD44, CD54 purchased from Boston. After washing thrice in phosphate-buffered saline (PBS), cells were reacted with secondary antibodies and then coloration with Bromine chlorine benzpyrrole phosphonic /nitroblue tetrazolium (BCTP/NBT). After blocked with block fluid, cells were incubated with monoclonal antibodies Brdu for 40 minutes. Then reacted with secondary antibodies and counterstain with BAD. MTT testMSCs culture was started at 1000 cells per well in 96-well tissue culture plates. Culture medium was IMDM supplemented with "Qiangjijianli" fluid, pooled serum consist of Qiangjijianli and tortoise, tortoise plastron, 5-azacytidine or blank serum. After two days' incubation, the MTT test was performed as routinely described. Biokinetics reader detected the values of absorbance at 490nm. All the data were expressed at mean±standard deviation (SD) . Independent-samples t test was carried out using SPSS11. 5 for windows software. Cell differentiation derivationThe differentiation ability of the MSCs was assessed at the third and tenth passages. The inducible conditions were based on the MSCs from bone marrow. The OS cells were seeded in 24-well plate (lOOOcells/well) and cultured in 5% co2 37°C for 3 ,7 or 14 days. The cell medium was discarded. The wells were washed with phosphate buffered saline (PBS), and lml serum-free medium containing medical serum at different final concentration. The samples were divided into different groups, either with serum-free medium, or with culture medium containing medicine. Cell immunohistochemistry assay
The fixed cells were harvested from tissue culture flasks. Aliquots were incubated for one night at refrigerator 4°C with m onoclonal antibodies against desmin or myosin purchased from Boston. After washing thrice in phosphate-buffered saline (PBS), cells were reacted with secondary antibodies and then coloration with BAD and counterstain with hematoxylin. Results:1. Theory researchThe study on the doctrine of "the correlation of five viscera" refered to many aspects involved in literature, clinic and experiment. As one of well-provided subsystem of "the correlation of five viscera" , predominant curative effect are showed on the treament of paralysis according the theory of "spleen-kidney related" . Internal empirical study report about the theory of "spleen-kidney reated" these year are finded one after another. But there are no study on Mesenchymal stem cells.2. Empirical study: Isolation and culture of MSCsRound cells appeared first in primary culture, then the cell were set in culture of incubation, spindle-like cells adherent to the Petri dishes appeared, followed by formation of colonies at day 3-7 and an adherent minelayer around 7-9 days. A variety of antibodies reacting with different cell lineages were used to investigate the surface antigenic markers present on MSCs. The results showed that MSCs were uniformly positive for CD44, CD54. Multiplication effect on MSCsMultiplication effect can be produced by variable dose of"Qiangjijianli" fluid, which is symbiosis with concentration. The similar result is shown on tortoise plastron. Pooled serum consists of"Qiangjijianli" and tortoise plastron have better effect on Multiplication of MSCs. Effect on differentiation of MSCs"Qiangjijianli" fluid, pooled serum consists of "Qiangjijianli" and tortoise plastron, 5-azacytidine could induce MSCs into myoblast. The differentiation of induced cells was identified by immunohistochemistry. The rate of positive cells didn' t have strong relation with concentration.Conclusion:
The doctrine of "the correlation of five viscera" is one of important ingredent of contemporary Chinese medicine academic. It showed that the therapeutic principle of Flaccidity syndrome is invigorated the spleen and kidney together. "Qiangjijianli" serum of Physiological dose can cause evident multiplication effect, the effect increasing with concentration. Pooled serum consists of "Qiangjijianli" and tortoise plastron with proper ratio have better effect on MSCs than the others. It showed that invigorate the spleen and kidney together are better than invigorate the spleen or kidney alone. "Qiangjijianli" fluid, pooled serum consists of "Qiangjijianli" and tortoise plastron, 5-azacytidine could induce MSCs into myoblast. It showed that the rat MSCss could be induced into myoblast cells by invigorating the spleen or invigorating the spleen and kidney together.
对五脏相关理论进行梳理和探讨,在此基础上,对子系统“脾肾相关”理论进行更为深入的研究,提出“骨髓间充质干细胞(MSCs)为脾肾相关细胞层次表达的存在形式”的工作假说,探讨补脾益肾法对大鼠MSCs增殖和分化的影响,从微观角度和干细胞水平研究脾肾相关。
研究方法:
一、五脏相关理论探讨
(一)五脏相关学说的缘起:
1.五脏相关源流探讨:经考证,词义最接近、年代最早的文献是《素问·玉机真脏论》:“五脏相通,移皆有次”。古人通过阐述五脏功能互用和脏腑相合相通理论、气机升降出入、疾病的传变、五脏旁通之说、天人相应及药性归经理论体现五脏之气相通。
2.中医五脏相关学说的形成与发展:邓铁涛教授在几十年的临床实践中,把五行学说和脏腑学说结合起来,提出了“五脏相关”学说。
(1)理论研究继续深化:90年代末本世纪初,邓铁涛教授指导两位博士研究生赵氏、郑氏对五脏相关学说科学内涵进行探讨。认为五脏相关学说是对中医经典理论五行学说的继承和发展,具有跨学科的特点。
(2)五脏相关学说临床研究:五脏相关临床研究是目前研究的重点,主要有两个层次的研究,一是五脏相关总体层面的研究,二是深入到子系统如两脏相关的研究。面上的研究显示五脏相关理论已经广泛应用于临床各个系统疾病的治疗,与此同时,各个子系统的研究也逐渐开展。目前,以两两相关的研究较多。如脾(胃)肾相关,心脾(胃)相关,肝脾(胃)相关,胆胃相关,肺脾(胃)相关,心肺相关,心胆相关,心肾相关,肺肾相关,肝肾相关,肝肺相关等。
(3)五脏相关学说实验研究:部分学者进行了实验研究,揭示了五脏中的肝、脾(这里指空肠绒毛)、肾之间的相生、相克关系,“肺肾相关”的物质基础,中医“心肾相关”理论的科学内涵等。
(二)脾肾相关子系统研究:
中医脾肾相关子系统以阐发脾和肾相关的生理功能、病理变化机理及其证治方药为中心内容,体现了脏腑同病病机和整体观念。与传统的中医理论比较,脾肾相关吸取了五行学说中“土制水”的精神内核,并综合了历代医家对脾肾相互关系的不断发展,而将二者之间动态的、错综复杂的相互关系用“相关”来概括。近年来,国内有
Objective:This topic discussed and systematized the theory of "spleen-kidney related" on the basic of literature. Which was one of subsystem of "the doctrine of the correlation of five viscera" put forward by Professor Tietao Deng, the famous TCM scientist of the contemporary. We would induce the frequent type and medication rule of Flaccidity syndrome in clinical part. While in experimental part, we approach the effect on multiplication and differentiation of Mesenchymal stem cells (MSCs) resulting from "Qiangjijianli" fluid for oral use. On the other hand, we' d like to discuss the mechanism of action of the medicine in curing Flaccidity syndrome.Methods:1. Theory researchThe original literature of the doctrine of the correlation of five viscera is "plain questions" .The ancients explain the communication of QI of five ZANG-organs. The theory are put forward by Professor tietao deng on the basic of clinical practice, who integrate theory of five-element and zang-fu organs. The theory are developed by his students, while are widely studied on clinical application and empirical study. one of subsystem of "the doctrine of the correlation of five viscera" was the theory of spleen-kidney related.2. Empirical study: Isolation and culture of MSCsHeparinized bone marrow was obtained from SD rat. The marrow samples were diluted with Is cove' s modified Dovecot' s medium (IMDM) containing 5% fetal bovine serum (FBS) and 5% horse serum (HS). The mononuclear cells were prepared by gradient centrifugation at 2000rpm. The cells were washed, counted and
plated at 10000 cells/75cm2 on Petri dishes in IMDM supplemental with 5% FBS and 5% HS which was selected for proper recovery and expansion of MSCs. The cells were incubated in a humidified incubator in 5% co2 at 37°C. Medium was replaced and the cells in suspension were removed at 48 hours and every three days thereafter. MSCs formed as confluent layers were detached by treatment with 0.25% trypsin(sigma) and seeded into cultural flasks. MSCs after the third subculture were used for the described experiments. Analysis of cell surface moleculesThe fixed cells were harvested from tissue culture flasks. Aliquots were incubated for one night at refrigerator 4°C with Multiclone antibody against rabbit CD44, CD54 purchased from Boston. After washing thrice in phosphate-buffered saline (PBS), cells were reacted with secondary antibodies and then coloration with Bromine chlorine benzpyrrole phosphonic /nitroblue tetrazolium (BCTP/NBT). After blocked with block fluid, cells were incubated with monoclonal antibodies Brdu for 40 minutes. Then reacted with secondary antibodies and counterstain with BAD. MTT testMSCs culture was started at 1000 cells per well in 96-well tissue culture plates. Culture medium was IMDM supplemented with "Qiangjijianli" fluid, pooled serum consist of Qiangjijianli and tortoise, tortoise plastron, 5-azacytidine or blank serum. After two days' incubation, the MTT test was performed as routinely described. Biokinetics reader detected the values of absorbance at 490nm. All the data were expressed at mean±standard deviation (SD) . Independent-samples t test was carried out using SPSS11. 5 for windows software. Cell differentiation derivationThe differentiation ability of the MSCs was assessed at the third and tenth passages. The inducible conditions were based on the MSCs from bone marrow. The OS cells were seeded in 24-well plate (lOOOcells/well) and cultured in 5% co2 37°C for 3 ,7 or 14 days. The cell medium was discarded. The wells were washed with phosphate buffered saline (PBS), and lml serum-free medium containing medical serum at different final concentration. The samples were divided into different groups, either with serum-free medium, or with culture medium containing medicine. Cell immunohistochemistry assay
The fixed cells were harvested from tissue culture flasks. Aliquots were incubated for one night at refrigerator 4°C with m onoclonal antibodies against desmin or myosin purchased from Boston. After washing thrice in phosphate-buffered saline (PBS), cells were reacted with secondary antibodies and then coloration with BAD and counterstain with hematoxylin. Results:1. Theory researchThe study on the doctrine of "the correlation of five viscera" refered to many aspects involved in literature, clinic and experiment. As one of well-provided subsystem of "the correlation of five viscera" , predominant curative effect are showed on the treament of paralysis according the theory of "spleen-kidney related" . Internal empirical study report about the theory of "spleen-kidney reated" these year are finded one after another. But there are no study on Mesenchymal stem cells.2. Empirical study: Isolation and culture of MSCsRound cells appeared first in primary culture, then the cell were set in culture of incubation, spindle-like cells adherent to the Petri dishes appeared, followed by formation of colonies at day 3-7 and an adherent minelayer around 7-9 days. A variety of antibodies reacting with different cell lineages were used to investigate the surface antigenic markers present on MSCs. The results showed that MSCs were uniformly positive for CD44, CD54. Multiplication effect on MSCsMultiplication effect can be produced by variable dose of"Qiangjijianli" fluid, which is symbiosis with concentration. The similar result is shown on tortoise plastron. Pooled serum consists of"Qiangjijianli" and tortoise plastron have better effect on Multiplication of MSCs. Effect on differentiation of MSCs"Qiangjijianli" fluid, pooled serum consists of "Qiangjijianli" and tortoise plastron, 5-azacytidine could induce MSCs into myoblast. The differentiation of induced cells was identified by immunohistochemistry. The rate of positive cells didn' t have strong relation with concentration.Conclusion:
The doctrine of "the correlation of five viscera" is one of important ingredent of contemporary Chinese medicine academic. It showed that the therapeutic principle of Flaccidity syndrome is invigorated the spleen and kidney together. "Qiangjijianli" serum of Physiological dose can cause evident multiplication effect, the effect increasing with concentration. Pooled serum consists of "Qiangjijianli" and tortoise plastron with proper ratio have better effect on MSCs than the others. It showed that invigorate the spleen and kidney together are better than invigorate the spleen or kidney alone. "Qiangjijianli" fluid, pooled serum consists of "Qiangjijianli" and tortoise plastron, 5-azacytidine could induce MSCs into myoblast. It showed that the rat MSCss could be induced into myoblast cells by invigorating the spleen or invigorating the spleen and kidney together.
引文
[1] 日本·山田业广.素问次注集疏[M],北京:学苑出版社,2004:451
[2] 明·马莳.黄帝内经素问注证发微[M],北京:学苑出版社,2003:197
[3][5] 明·李梃.医学入门[M],北京:中国中医药出版社,1995:3、72
[4] 清·汪昂《本草备要·药性总义》[M],北京:中国中医药出版社,1998:4
[6][7] 清·何梦瑶.医碥[M],广州:广东科技出版社,1981:7、9
[8] 邓铁涛.邓铁涛医集[M].北京:人民卫生出版社,1995,156-165
[9] 刘小斌,刘友章.邓铁涛五脏相关学说略介[J].上海中医药杂志,2002;(7):36—37
[10] 徐志伟.李俊德,邓铁涛学术思想研究[M].北京:华夏出版社,2001.103.
[11] 邱仕君.在基础理论与临床医学之间——对邓铁涛教授五脏相关学说的理论思考.邓铁涛学术思想研究(Ⅱ),北京:华夏出版社,2004:41—46
[12] 张筱文.脏腑相关治则探讨[J].辽宁中医杂志,1995,22(6):245-247
[13] 徐志伟.李俊德,邓铁涛学术思想研究[M].北京:华夏出版社,2001.101～106
[14] 马淑然,刘燕池,郭霞珍等,中医五脏是人体生理调控模板[J].中国中医基础医学杂志,2002,(11):1
[15] 周福生,许仁杰,黄纯美。“三位一体”辨证与邓老“五脏相关理论”的研究。邓铁涛学术思想研究(Ⅱ),67-71
[16] 李振吉.中国中医药科技成果获奖项目汇编[M].北京:中医古籍出版社,2003.37
[17] 李振吉.中国中医药科技成果获奖项目汇编[M].北京:中医古籍出版社,2003.17
[18] 刘小斌,刘友章,王清海,等.五脏相关理论学说临床基础调研——附1200例病案资料分析[J].中医药学刊,2004;22(2):211—213
[19] 刘小斌.邓铁涛教授对五脏相关理论的临床应用[J].中国中医基础医学杂志,2001;7(11):73—74
[20] 刘英君.张腾.潘玉珍等,冠心病五脏相关兼治探略[J].中医药学报,1998:(1):6～7
[21] 刘小斌,刘友章.邓铁涛教授抢救重症肌无力危象的方法与思路[J].河南中医,2004;24(1):18—19
[22] 邓铁涛,刘友章,刘小斌.中西医结合抢救26例重症肌无力危象[J].广州中医药大学学报,2004;21(5):344—348
[23] 车宇光.从“五脏相关说”论治慢性心力衰竭[J].中国中医急症,2003;12(2):181
[24] 谢雨君.陈镜合教授治疗无症状性心肌缺血经验介绍[J].新中医,2002;34(11):9-10
[25] 刘英君,张腾,潘玉珍,等.冠心病五脏相关兼治探略[J],中医药学报,1998; (1):6—7
[26] 贺敬波,陈捷,朱秋艳.从肝论治冠心病的理论探讨[J].深圳中西医结合杂志,2003;13(1):7-8
[27] 尹宗强.浅谈五脏相关理论在病毒性肝炎治疗中的应用[J].中西医结合肝病杂志,1998;8 (SS):17-18
[28] 韩立国,邢召君.从脏腑相关论胃脘痛的治法[J].时珍国医国药,2004;15(10):712
[29] 谢桂权.尿毒症中医治疗整体观探析[J].新中医,2002;34(10):3-5
[30] 陈遂生.肺心病证治5则[J].中医研究,1995;8(2):39-41
[31] 魏华,路洁.路志正教授治疗甲状腺机能亢进症的用药经验[J].广州中医药大学学报,2004;21(5):407-409
[32] 谭抗美.脏腑相关理论诊治女性更年期综合证临床应用研究(附61例分析)[J].中国临床医药研究杂志,2003;(91):8933—8934
[33] 宋述财,刘小斌,许华.失眠与五脏相关刍议[J].中国医学理论与实践,2003;(3):367-364
[34] 曹红霞.补心通脉片治疗老年顽固性失眠疗效观察[J].中华医学与写作杂志,2002;9(9):16
[35] 葛淑芬,等.顽固失眠从肝胆论治[J].上海中医药杂志,1997:(7):13
[36] 李景.失眠证从脾胃论治[J].中国中医基础医学杂志,2002;8(1):28
[37] 毛文灏,等.老年慢性肺心病急性发作期神经精神症状鉴别诊断[J].医师进修杂志,1996:19(8):22
[38] 吕春芳,等.吕同杰治疗顽固性失眠经验[J].山东中医杂志,2000;17(5):300
[39] 王文奎.中医论治艾滋病[J].中国工程科学,2004:6(1):44—47
[40] 边玉麟.五脏相关治未病临床浅探[J].安徽中医临床杂志,2002:14(2):145
[41] 吴焕林,阮新民,张敏州,等.冠心病冠状动脉搭桥术后患者的中医证候分析[J].广州中医药大学大学学报,2000;17(3):228—229
[42] 张庆宏,周福生.析“心胃相关”[J].云南中医学院学报,2001;24(2):39
[43] 贾风新,李桂.从脾胃的生理特点探讨消化性溃疡的组方结构[J].中国中医基础医学杂志,2001;7(5):43-4
[44] 肖会泉,罗日永,吴婉芬.肝脾相关理论在慢性肝炎治疗中的运用[J].邓铁涛学术思想研究(Ⅱ),169—175
[45] 范廷校.肝胃相关理论浅识[J].福建中医药,2003;34(1):48-49
[46] 杨雨田,武俊青.胆石症患者胃电图特征与胆胃相关说初探[J].山西中医,1998;14(3):34-35
[47] 徐虹,顾庆华.从胆胃相关探讨胆囊炎与胆汁返流性胃炎的关系[J].中医药研究,1996;(5):17
[48] 李卫芳.胆胃同治、疏活共用治疗慢性胆病经验[J].吉林中医药,2004;24(11):16-17
[49] 张筱文.论胆胃相关与同治[J].中华实用中西医杂志,2004;17(2):238
[50] 张挹芳.肺脾理论与过敏性疾病[J].上海中医药杂志,1997;(9):16-17
[51] 刘小虹,刘琼.培土生金法在慢性阻塞性肺疾病缓解期的应用研究[J].新中医,2002:34(10):18—19
[52] 林沁臻.浅谈肺系疾病从脾论治[J].湖北中医杂志,2004:26(2):21
[53] 段延萍,周杰,高连印.肺胃相关论探析[J].陕西中医,2003;24(12):1104-1106
[54] 郑杨,林琳,张静.心肺相关理论探讨[J].辽宁中医学院学报,2002;4(2):84-85
[55] 刘学法.从心肺相关论冠心病的证治[J].光明中医,1999;14(4):12-13
[56] 王宗柱.略论“胆气通于心” [J].陕西中医学院学报.1999;22(6):1-2
[57] 陆曙.心胆相关理论探略[J].陕西中医,1992;13(7):304-306
[58] 郝明慧,王者仁,周淑华.心肾相关理论对治疗心衰的指导作用[J].实用中医内科杂志,1996;10(4):21
[59] 刘勤社.心肾相关实质初探[J].陕西中医,1991;12(4):163-164
[60] 何灵芝,王晓亚.从慢性肾衰尿毒症肺功能变化探讨肺肾关系[J].浙江中医杂志,1998;33(6):273
[61] 付修文.肝肾相关与慢性肝病[J].北京中医,1995;(2):48-50
[62] 武维屏,崔红生.试论支气管哮喘从肝论治的生理病理学基础[J].中国中医基础医学杂志,2002;8(10):7-8
[63] 张丽娟.有关实验性肝硬变脏腑相关性的探讨.国外医学·中医中药分册,1997;19(6):32
[64] 许辉,陈松,李家邦等.特异性免疫应答在肝郁证与脾虚证及相互关系的初步研究[J].中国医师杂志,2002;(8):829-831
[65] 郑爱华,李家邦,蔡光先,等.柴胡疏肝散、四君子汤对肝郁、脾虚病人Th细胞分化信号蛋白PKC表达的影响[J].贵阳中医学院学报,2003;25(4):14-17
[66] 王淑玲.肺肾相关的物质基础[J].中国中医基础医学杂志,2002;(4):9
[67] 王淑玲,宗全和.中西医结合探讨“肺肾相关”[J].中国医药学报,2002;17(4):221-223
[68] 陈洁文,罗荣敬,胡维安.中医“心肾相关”理论的实验研究[J].广州中医学院学报,1991;(2/3):113-118
[69] 刘成丽.中医脾肾相关学说源流探讨[J].中医药信息,2003:20(6):1-3
[70] 屠伯言.中医辨证治疗糖尿病肾病13例临床分析[J].中医杂志,1984;25(4),39
[71] 陶庆文,阎小萍.脾肾相关理论在WANG痹治疗中的应用[J].中国临床医生,2002;30(8):57-58
[72] 唐光富.胃肾相关理论价值新识[J].广西中医学院学报,2000;17(3):15-16
[73] 吕爱平,李德新.脾肾虚证与生物膜关系的研究进展[J].辽宁中医杂志,2002;27(10):479-480
[74] 吕爱平,李德新,崔家鹏,等.脾肾相关的分子生物学基础-脾肾阳虚证模型大鼠自由基损伤的比较研究[J].辽宁中医杂志,2001:28(3):189-190
[75] 吕爱平,林庶茹,李德新,等.脾肾阳虚模型大鼠肝细胞线粒体磷脂组分变化的比较研究[J].中医药学刊,2003;21(1):78,83
[76] 孙爱华,等.肾与脾关系物质基础的实验观察[J].中医杂志.1998;39(1):47～49
[77] 高守泉,等.脾肾相关性进行的实验研究[J].湖南中医药导报.1998;4(12):26～27.
[78] 陈芝喜,刘小斌,周名璐,等.补脾补肾药对脾虚大鼠细胞因子水平的影响[J].中国临床康复, 2004;8(24):5186-5188
[79] 陈芝喜,刘小斌,周名璐,等.补脾补肾药对脾虚大鼠环核苷酸及甲状腺激素水平的影响[J].放射免疫杂志,2003;16(5):278—280
[80] 周英,梁国珍,罗颂平.脾虚和肾虚雌性大鼠生育能力的实验观察[J].广州中医药大学学报,1998:(3):195-199
[81] 刘东辉,雍翠云,王汝俊,等.强肌健力口服液主要药效学研究[J].中国实验方剂学杂志,2001:7(3):44-46
[82] 李顺民.强肌健力胶囊对烟碱型乙酰胆碱受体代谢的影响[J].中国中西医结合杂志,1997;17(4):230-231
[83] 李顺民.强肌健力胶囊治疗重症肌无力免疫学分析[J].广州中医学院学报,1992;9(2):65-67
[84] 刘东辉,雍翠云,玉汰俊.强肌健力口服液主要药效学研究[J].中国实验方剂学杂志,2001:7(3):44-46
[85] 杨莉,王冬梅,洪欣.成人骨髓间充质干细胞体外扩增和定向诱导分化为骨和软骨细胞的研究[J].自然科学进展,2001;11(10):1062-1067
[86] 张进,徐志伟.补肾法诱导间充质干细胞向神经方向分化研究[J].现代医院,2004;4(9):15-17
[87] 张勇,邹仲敏,郭朝华,等.MyoD基因诱导骨髓间充质干细胞分化为成肌细胞的实验研究[J].第三军医大学学报.2002;24(12):1425-1426
[88] 胡华,梁志清,徐惠成,等.大鼠骨髓间充质细胞向成肌细胞诱导分化的研究[J].重庆医学.2004;33(5):675-677
[89] 张进,徐志伟,杜少辉,等.“精”学说与干细胞辨识[J].中医药学刊,2004;22(7):1198-1200
[90] 陈东风,杜少辉,李伊为,等.龟板含药血清体外诱导成年大鼠骨髓问充质干细胞分化为神经元[J].广州中医药大学学报,2003;20(3):224-226
[91] 周健洪,陈东风,黎晖,等.龟板含药血清对大鼠骨髓间质干细胞体外增殖的影响[J].广州中医药大学学报.2005:22(1):35-38
[92] 王劲,郭朝华,张勇,等.5-氮杂胞苷诱导间充质干细胞分化为成肌细胞的实验研究[J].中华创伤杂志.2003;19(4):207-211
[93] Jennings CG.Expression of the myogenic gene MRF_4 during xenopus development.Dev Riol,1992;151:319-332
[94] 郑奇军,蔡振杰,俞世强,等.骨髓间质干细胞体外转化为心肌细胞的实验研究[J].第四军医大学学报.2002;23(20):1871-1873
[95] 林先和,李隆.5-氮杂胞苷诱导小鼠骨髓间充质干细胞分化过程中PPARγ表达及其作用[J].第三军医大学学报,2004,26(6):526-529
[96] 王劲,郭朝华,张勇.成肌细胞增殖/分化的信号转导研究[J].生命的化学,2002,22(1):18-20
[2] 明·马莳.黄帝内经素问注证发微[M],北京:学苑出版社,2003:197
[3][5] 明·李梃.医学入门[M],北京:中国中医药出版社,1995:3、72
[4] 清·汪昂《本草备要·药性总义》[M],北京:中国中医药出版社,1998:4
[6][7] 清·何梦瑶.医碥[M],广州:广东科技出版社,1981:7、9
[8] 邓铁涛.邓铁涛医集[M].北京:人民卫生出版社,1995,156-165
[9] 刘小斌,刘友章.邓铁涛五脏相关学说略介[J].上海中医药杂志,2002;(7):36—37
[10] 徐志伟.李俊德,邓铁涛学术思想研究[M].北京:华夏出版社,2001.103.
[11] 邱仕君.在基础理论与临床医学之间——对邓铁涛教授五脏相关学说的理论思考.邓铁涛学术思想研究(Ⅱ),北京:华夏出版社,2004:41—46
[12] 张筱文.脏腑相关治则探讨[J].辽宁中医杂志,1995,22(6):245-247
[13] 徐志伟.李俊德,邓铁涛学术思想研究[M].北京:华夏出版社,2001.101～106
[14] 马淑然,刘燕池,郭霞珍等,中医五脏是人体生理调控模板[J].中国中医基础医学杂志,2002,(11):1
[15] 周福生,许仁杰,黄纯美。“三位一体”辨证与邓老“五脏相关理论”的研究。邓铁涛学术思想研究(Ⅱ),67-71
[16] 李振吉.中国中医药科技成果获奖项目汇编[M].北京:中医古籍出版社,2003.37
[17] 李振吉.中国中医药科技成果获奖项目汇编[M].北京:中医古籍出版社,2003.17
[18] 刘小斌,刘友章,王清海,等.五脏相关理论学说临床基础调研——附1200例病案资料分析[J].中医药学刊,2004;22(2):211—213
[19] 刘小斌.邓铁涛教授对五脏相关理论的临床应用[J].中国中医基础医学杂志,2001;7(11):73—74
[20] 刘英君.张腾.潘玉珍等,冠心病五脏相关兼治探略[J].中医药学报,1998:(1):6～7
[21] 刘小斌,刘友章.邓铁涛教授抢救重症肌无力危象的方法与思路[J].河南中医,2004;24(1):18—19
[22] 邓铁涛,刘友章,刘小斌.中西医结合抢救26例重症肌无力危象[J].广州中医药大学学报,2004;21(5):344—348
[23] 车宇光.从“五脏相关说”论治慢性心力衰竭[J].中国中医急症,2003;12(2):181
[24] 谢雨君.陈镜合教授治疗无症状性心肌缺血经验介绍[J].新中医,2002;34(11):9-10
[25] 刘英君,张腾,潘玉珍,等.冠心病五脏相关兼治探略[J],中医药学报,1998; (1):6—7
[26] 贺敬波,陈捷,朱秋艳.从肝论治冠心病的理论探讨[J].深圳中西医结合杂志,2003;13(1):7-8
[27] 尹宗强.浅谈五脏相关理论在病毒性肝炎治疗中的应用[J].中西医结合肝病杂志,1998;8 (SS):17-18
[28] 韩立国,邢召君.从脏腑相关论胃脘痛的治法[J].时珍国医国药,2004;15(10):712
[29] 谢桂权.尿毒症中医治疗整体观探析[J].新中医,2002;34(10):3-5
[30] 陈遂生.肺心病证治5则[J].中医研究,1995;8(2):39-41
[31] 魏华,路洁.路志正教授治疗甲状腺机能亢进症的用药经验[J].广州中医药大学学报,2004;21(5):407-409
[32] 谭抗美.脏腑相关理论诊治女性更年期综合证临床应用研究(附61例分析)[J].中国临床医药研究杂志,2003;(91):8933—8934
[33] 宋述财,刘小斌,许华.失眠与五脏相关刍议[J].中国医学理论与实践,2003;(3):367-364
[34] 曹红霞.补心通脉片治疗老年顽固性失眠疗效观察[J].中华医学与写作杂志,2002;9(9):16
[35] 葛淑芬,等.顽固失眠从肝胆论治[J].上海中医药杂志,1997:(7):13
[36] 李景.失眠证从脾胃论治[J].中国中医基础医学杂志,2002;8(1):28
[37] 毛文灏,等.老年慢性肺心病急性发作期神经精神症状鉴别诊断[J].医师进修杂志,1996:19(8):22
[38] 吕春芳,等.吕同杰治疗顽固性失眠经验[J].山东中医杂志,2000;17(5):300
[39] 王文奎.中医论治艾滋病[J].中国工程科学,2004:6(1):44—47
[40] 边玉麟.五脏相关治未病临床浅探[J].安徽中医临床杂志,2002:14(2):145
[41] 吴焕林,阮新民,张敏州,等.冠心病冠状动脉搭桥术后患者的中医证候分析[J].广州中医药大学大学学报,2000;17(3):228—229
[42] 张庆宏,周福生.析“心胃相关”[J].云南中医学院学报,2001;24(2):39
[43] 贾风新,李桂.从脾胃的生理特点探讨消化性溃疡的组方结构[J].中国中医基础医学杂志,2001;7(5):43-4
[44] 肖会泉,罗日永,吴婉芬.肝脾相关理论在慢性肝炎治疗中的运用[J].邓铁涛学术思想研究(Ⅱ),169—175
[45] 范廷校.肝胃相关理论浅识[J].福建中医药,2003;34(1):48-49
[46] 杨雨田,武俊青.胆石症患者胃电图特征与胆胃相关说初探[J].山西中医,1998;14(3):34-35
[47] 徐虹,顾庆华.从胆胃相关探讨胆囊炎与胆汁返流性胃炎的关系[J].中医药研究,1996;(5):17
[48] 李卫芳.胆胃同治、疏活共用治疗慢性胆病经验[J].吉林中医药,2004;24(11):16-17
[49] 张筱文.论胆胃相关与同治[J].中华实用中西医杂志,2004;17(2):238
[50] 张挹芳.肺脾理论与过敏性疾病[J].上海中医药杂志,1997;(9):16-17
[51] 刘小虹,刘琼.培土生金法在慢性阻塞性肺疾病缓解期的应用研究[J].新中医,2002:34(10):18—19
[52] 林沁臻.浅谈肺系疾病从脾论治[J].湖北中医杂志,2004:26(2):21
[53] 段延萍,周杰,高连印.肺胃相关论探析[J].陕西中医,2003;24(12):1104-1106
[54] 郑杨,林琳,张静.心肺相关理论探讨[J].辽宁中医学院学报,2002;4(2):84-85
[55] 刘学法.从心肺相关论冠心病的证治[J].光明中医,1999;14(4):12-13
[56] 王宗柱.略论“胆气通于心” [J].陕西中医学院学报.1999;22(6):1-2
[57] 陆曙.心胆相关理论探略[J].陕西中医,1992;13(7):304-306
[58] 郝明慧,王者仁,周淑华.心肾相关理论对治疗心衰的指导作用[J].实用中医内科杂志,1996;10(4):21
[59] 刘勤社.心肾相关实质初探[J].陕西中医,1991;12(4):163-164
[60] 何灵芝,王晓亚.从慢性肾衰尿毒症肺功能变化探讨肺肾关系[J].浙江中医杂志,1998;33(6):273
[61] 付修文.肝肾相关与慢性肝病[J].北京中医,1995;(2):48-50
[62] 武维屏,崔红生.试论支气管哮喘从肝论治的生理病理学基础[J].中国中医基础医学杂志,2002;8(10):7-8
[63] 张丽娟.有关实验性肝硬变脏腑相关性的探讨.国外医学·中医中药分册,1997;19(6):32
[64] 许辉,陈松,李家邦等.特异性免疫应答在肝郁证与脾虚证及相互关系的初步研究[J].中国医师杂志,2002;(8):829-831
[65] 郑爱华,李家邦,蔡光先,等.柴胡疏肝散、四君子汤对肝郁、脾虚病人Th细胞分化信号蛋白PKC表达的影响[J].贵阳中医学院学报,2003;25(4):14-17
[66] 王淑玲.肺肾相关的物质基础[J].中国中医基础医学杂志,2002;(4):9
[67] 王淑玲,宗全和.中西医结合探讨“肺肾相关”[J].中国医药学报,2002;17(4):221-223
[68] 陈洁文,罗荣敬,胡维安.中医“心肾相关”理论的实验研究[J].广州中医学院学报,1991;(2/3):113-118
[69] 刘成丽.中医脾肾相关学说源流探讨[J].中医药信息,2003:20(6):1-3
[70] 屠伯言.中医辨证治疗糖尿病肾病13例临床分析[J].中医杂志,1984;25(4),39
[71] 陶庆文,阎小萍.脾肾相关理论在WANG痹治疗中的应用[J].中国临床医生,2002;30(8):57-58
[72] 唐光富.胃肾相关理论价值新识[J].广西中医学院学报,2000;17(3):15-16
[73] 吕爱平,李德新.脾肾虚证与生物膜关系的研究进展[J].辽宁中医杂志,2002;27(10):479-480
[74] 吕爱平,李德新,崔家鹏,等.脾肾相关的分子生物学基础-脾肾阳虚证模型大鼠自由基损伤的比较研究[J].辽宁中医杂志,2001:28(3):189-190
[75] 吕爱平,林庶茹,李德新,等.脾肾阳虚模型大鼠肝细胞线粒体磷脂组分变化的比较研究[J].中医药学刊,2003;21(1):78,83
[76] 孙爱华,等.肾与脾关系物质基础的实验观察[J].中医杂志.1998;39(1):47～49
[77] 高守泉,等.脾肾相关性进行的实验研究[J].湖南中医药导报.1998;4(12):26～27.
[78] 陈芝喜,刘小斌,周名璐,等.补脾补肾药对脾虚大鼠细胞因子水平的影响[J].中国临床康复, 2004;8(24):5186-5188
[79] 陈芝喜,刘小斌,周名璐,等.补脾补肾药对脾虚大鼠环核苷酸及甲状腺激素水平的影响[J].放射免疫杂志,2003;16(5):278—280
[80] 周英,梁国珍,罗颂平.脾虚和肾虚雌性大鼠生育能力的实验观察[J].广州中医药大学学报,1998:(3):195-199
[81] 刘东辉,雍翠云,王汝俊,等.强肌健力口服液主要药效学研究[J].中国实验方剂学杂志,2001:7(3):44-46
[82] 李顺民.强肌健力胶囊对烟碱型乙酰胆碱受体代谢的影响[J].中国中西医结合杂志,1997;17(4):230-231
[83] 李顺民.强肌健力胶囊治疗重症肌无力免疫学分析[J].广州中医学院学报,1992;9(2):65-67
[84] 刘东辉,雍翠云,玉汰俊.强肌健力口服液主要药效学研究[J].中国实验方剂学杂志,2001:7(3):44-46
[85] 杨莉,王冬梅,洪欣.成人骨髓间充质干细胞体外扩增和定向诱导分化为骨和软骨细胞的研究[J].自然科学进展,2001;11(10):1062-1067
[86] 张进,徐志伟.补肾法诱导间充质干细胞向神经方向分化研究[J].现代医院,2004;4(9):15-17
[87] 张勇,邹仲敏,郭朝华,等.MyoD基因诱导骨髓间充质干细胞分化为成肌细胞的实验研究[J].第三军医大学学报.2002;24(12):1425-1426
[88] 胡华,梁志清,徐惠成,等.大鼠骨髓间充质细胞向成肌细胞诱导分化的研究[J].重庆医学.2004;33(5):675-677
[89] 张进,徐志伟,杜少辉,等.“精”学说与干细胞辨识[J].中医药学刊,2004;22(7):1198-1200
[90] 陈东风,杜少辉,李伊为,等.龟板含药血清体外诱导成年大鼠骨髓问充质干细胞分化为神经元[J].广州中医药大学学报,2003;20(3):224-226
[91] 周健洪,陈东风,黎晖,等.龟板含药血清对大鼠骨髓间质干细胞体外增殖的影响[J].广州中医药大学学报.2005:22(1):35-38
[92] 王劲,郭朝华,张勇,等.5-氮杂胞苷诱导间充质干细胞分化为成肌细胞的实验研究[J].中华创伤杂志.2003;19(4):207-211
[93] Jennings CG.Expression of the myogenic gene MRF_4 during xenopus development.Dev Riol,1992;151:319-332
[94] 郑奇军,蔡振杰,俞世强,等.骨髓间质干细胞体外转化为心肌细胞的实验研究[J].第四军医大学学报.2002;23(20):1871-1873
[95] 林先和,李隆.5-氮杂胞苷诱导小鼠骨髓间充质干细胞分化过程中PPARγ表达及其作用[J].第三军医大学学报,2004,26(6):526-529
[96] 王劲,郭朝华,张勇.成肌细胞增殖/分化的信号转导研究[J].生命的化学,2002,22(1):18-20