双龙方干预骨髓间充质干细胞分化过程中代谢组学研究
|
摘要
双龙方是由丹参和人参两味药材所组成的中药复方配伍,具有“益气养血、活血通脉”的功能,适用于治疗冠心病、心绞痛和心肌梗死等心血管疾病。
骨髓间充质干细胞(MSCs)在一定条件之下定向分化为心肌细胞,对细胞移植治疗心肌梗死有重要的意义。本论文在代谢组学的层面上,研究中药复方双龙方对大鼠骨髓间充质干细胞的干预结果,寻找细胞代谢物在诱导过程中的变化。为此我们先对复方的组分配伍(总丹酚酸、人参总皂苷)及成分配伍(Sal B、Rb1)进行化学分析,通过HPLC/MS的方法确定了其中的11种化学成分,明确复方当中的化学组成。
然后又采用了离子对色谱法进行分离,离子阱质谱仪作检测,建立起分析MSCs代谢物的HPLC/MS方法,筛选出30种细胞代谢物,包括了核苷酸类、氨基酸类和糖代谢类,对其中的28种进行定量,并作了方法学考察。
在上述的基础上,进行了细胞代谢物组学研究。考察了干细胞在第0、10、20、30和35天的分化情况,比较空白组、5-aza诱导组、双龙方组分组和双龙方成分组之间的差别,得到了初步结果。双龙方组分组和5-aza诱导组中,有5种代谢物(AMP、Tau、ADP、Glu和Thr)在诱导第35天时浓度水平接近,说明这两个组具有相似疗效,与实验室前有的结果相符合。体现出中药复方配伍的必要性和科学性,在一定程度上具有与西药相似的治疗效果。结果对双龙方干预MSCs分化中代谢物的变化情况提供了参考。
Shuang Long Fang, which is composed of DanShen and JinShen, is a traditional Chinese medicine compound compatibitity. It is suitable for treating the cardiovascular disease such as coronary heart disease, angina and myocardial infarction.
Bone Marrow Mesenchymal Stem Cells(MSCs) can differentiate towards cardiomyocytes directly under certain conditions. This is a good news about treating the myocardial infarction by cell transplantation. The thesis is to find out the intervened result from MSCs of rats by the Chinese Traditional Medicine Shuang Long Fang on the level of metabolomics. So we can find out the change of cell metabolites caused by induction. First we have analyzed the component compatibility(Salvianolic acid and Ginsenosides) and composition compatibility(Sal B and Rb1) of the Shuang Long Fang compound. By using the HPLC/MS method, we confirmed 11 chemical compositions in it.
Then we used the Ion-pair chromatography and the Ion Trap mass spectrometry to separate and detect the metabolites of MSCs. We screened out 30 cell metabolites and 28 of them were quantified. Metabolites include nucleotides, amino acids and glycometabolism. The research of methodology is also carried out.
On the basis of these, we developed the study of metabolomics. The experiment obsevered the cell differentiation at the time point of 0, 10, 20, 30 and 35days. And a comparation of the Blank Group, 5-aza-induced Group, Shuang Long Fang component Group and Shuang Long Fang composition Group has been finished. The primary results have been found. We have found the concentration level of 5 metabolites(AMP, Tau, ADP, Glu,and Thr) is similar in the group of 5-aza-induced and Shuang Long Fang component, when the induction time reached 35 days. It is showed that this two groups have the similar curative effect, so this is coincident with the results concluded by our Laboratory before. So the compound compatibility of Traditional Chinese Medicine is necessary and scientific. The curative effect is similar to Western medicine to some extent. The result provided the references of the changing rules of metabolites in the intervention of MSCs by Shuang Long Fang.
骨髓间充质干细胞(MSCs)在一定条件之下定向分化为心肌细胞,对细胞移植治疗心肌梗死有重要的意义。本论文在代谢组学的层面上,研究中药复方双龙方对大鼠骨髓间充质干细胞的干预结果,寻找细胞代谢物在诱导过程中的变化。为此我们先对复方的组分配伍(总丹酚酸、人参总皂苷)及成分配伍(Sal B、Rb1)进行化学分析,通过HPLC/MS的方法确定了其中的11种化学成分,明确复方当中的化学组成。
然后又采用了离子对色谱法进行分离,离子阱质谱仪作检测,建立起分析MSCs代谢物的HPLC/MS方法,筛选出30种细胞代谢物,包括了核苷酸类、氨基酸类和糖代谢类,对其中的28种进行定量,并作了方法学考察。
在上述的基础上,进行了细胞代谢物组学研究。考察了干细胞在第0、10、20、30和35天的分化情况,比较空白组、5-aza诱导组、双龙方组分组和双龙方成分组之间的差别,得到了初步结果。双龙方组分组和5-aza诱导组中,有5种代谢物(AMP、Tau、ADP、Glu和Thr)在诱导第35天时浓度水平接近,说明这两个组具有相似疗效,与实验室前有的结果相符合。体现出中药复方配伍的必要性和科学性,在一定程度上具有与西药相似的治疗效果。结果对双龙方干预MSCs分化中代谢物的变化情况提供了参考。
Shuang Long Fang, which is composed of DanShen and JinShen, is a traditional Chinese medicine compound compatibitity. It is suitable for treating the cardiovascular disease such as coronary heart disease, angina and myocardial infarction.
Bone Marrow Mesenchymal Stem Cells(MSCs) can differentiate towards cardiomyocytes directly under certain conditions. This is a good news about treating the myocardial infarction by cell transplantation. The thesis is to find out the intervened result from MSCs of rats by the Chinese Traditional Medicine Shuang Long Fang on the level of metabolomics. So we can find out the change of cell metabolites caused by induction. First we have analyzed the component compatibility(Salvianolic acid and Ginsenosides) and composition compatibility(Sal B and Rb1) of the Shuang Long Fang compound. By using the HPLC/MS method, we confirmed 11 chemical compositions in it.
Then we used the Ion-pair chromatography and the Ion Trap mass spectrometry to separate and detect the metabolites of MSCs. We screened out 30 cell metabolites and 28 of them were quantified. Metabolites include nucleotides, amino acids and glycometabolism. The research of methodology is also carried out.
On the basis of these, we developed the study of metabolomics. The experiment obsevered the cell differentiation at the time point of 0, 10, 20, 30 and 35days. And a comparation of the Blank Group, 5-aza-induced Group, Shuang Long Fang component Group and Shuang Long Fang composition Group has been finished. The primary results have been found. We have found the concentration level of 5 metabolites(AMP, Tau, ADP, Glu,and Thr) is similar in the group of 5-aza-induced and Shuang Long Fang component, when the induction time reached 35 days. It is showed that this two groups have the similar curative effect, so this is coincident with the results concluded by our Laboratory before. So the compound compatibility of Traditional Chinese Medicine is necessary and scientific. The curative effect is similar to Western medicine to some extent. The result provided the references of the changing rules of metabolites in the intervention of MSCs by Shuang Long Fang.
引文
[1] Jessup M and Brozena S. Medical progress: Heart failure. N. Engl. J. Med. 2003, 348: 2007-2018
[2] Friedenstein A J, Chailakhyan R K and Gerasimov U V. Bone marrow osteogenic stem cells : invitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet 1987, 20: 263-272
[3] Jiang Y H, Jahagirdar B N, Reinhardt R L, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002, 418: 41-49
[4] Barrias C C, Ribeiro C C, Lamghari M, et al. Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres. J. Biomed. Mater. Res. Part A. 2005, 72A: 57-66
[5] Rickard D J, Sullivan T A, Shenker B J, et al. Induction Of Rapid Osteoblast Differentiation In Rat Bone-Marrow Stromal Cell-Cultures By Dexamethasone And Bmp-2. Dev. Biol. 1994, 161: 218-228
[6] Heino T J, Hentunen T A and Vaananen H K. Conditioned medium from osteocytes stimulates the proliferation of bone marrow mesenchymal stem cells and their differentiation into ostcoblasts. Exp.Cell Res. 2004, 294: 458-468
[7] Martin I, Muraglia A, Campanile G, et al. Fibroblast growth factor-2 supports ex vivo expansion and maintenance of osteogenic precursors from human bone marrow. Endocrinology 1997, 138: 4456-4462
[8] Sugiura F, Kitoh H and Ishiguro N. Osteogenic potential of rat mesenchymal stem cells after several passages. Biochem. Biophys. Res. Commun. 2004, 316: 233-239
[9] Caterson E J, Nesti L J, Li W J, et al. Three-dimensional cartilage formation by bone marrow-derived cells seeded ion polylactide/alginate amalgam. J. Biomed. Mater. Res. 2001, 57: 394-403
[10] Guo X M, Wang C Y, Zhang Y F, et al. Repair of large articular cartilage defects with implants of autologous mesenchymal stem cells seeded into beta-tricalcium phosphate in a sheep model. Tissue Eng. 2004, 10: 1818-1829
[11] Ferrari G, Cusella-De Angelis G, Coletta M, et al. Muscle regeneration by bone marrow derived myogenic progenitors. Science 1998, 279: 1528-1530
[12] Tomita S, Li R K, Weisel R D, et al. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation 1999, 100: 247-256 Suppl
[13] Makino S, Fukuda K, Miyoshi S, et al.Cardiomyocytes can be generated from marrow stromal cells in vitro. J. Clin. Invest. 1999, 103 (5): 697-705
[14] Ye NS (Ye, Neng-Sheng), Chen J (Chen, Jun), Luo GA (Luo, Guo-An), et al. Proteomic profiling of rat bone marrow mesenchymal stem cells induced by 5-azacytidine. Stem Cells and Development, 2006, 15 (5): 665-676
[15] Jeong JA, Lee Y, Lee W, et al. Proteomic analysis of the hydrophobic fraction of mesenchymal stem cells derived from human umbilical cord blood. Molecules and Cells, 2006, 22 (1): 36-43
[16] Wang DJ, Park JS, Chu JSF, et al. Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor beta 1 stimulation. Journal of Biological Chemistry 2004, 279 (42): 43725-43734
[17] Phinney DG, Baddoo M, Dutreil M, et al. Murine mesenchymal stem cells transplanted to the central nervous system of neonatal versus adult mice exhibit distinct engraftment kinetics and express receptors that guide neuronal cell migration. Stem Cells and Development, 2006, 15 (3): 437-447
[18] Forest D, Nishikawa R, Kobayashi H, et al. RNA expression in a cartilaginous fish cell line reveals ancient 3' noncoding regions highly conserved in vertebrates. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104 (4): 1224-1229
[19] Nicholson J K, Connelly J, Lindon J C, et al. Metabonomics:ap latform for studying drug toxicity and gene function. Nat Rev Drug Discov, 2002, 1 (2): 153-161
[20]冉小蓉,梁琼麟,罗国安等.代谢物组学的应用及进展.中成药2005, 27(4): 381-385
[21] Taylor J, King R D, Altmann T, et al. Application of metabolomics to plant genotype discrimination using statistics and machine learning. Bioinform, 2002, 18:S241-S248
[22] Vikram A, Prithiviraj B, Hamzehzarghani H, et al. Valatile metabolite profiling to discriminate diseases of Mc Intosh app le inoculated with fungal pathogens. J Sci FD Agric, 2004, 84(11): 1333-1340
[23] Manuel M, Yuen-Li C, Ursula M, et al. Proteomic and Metabolomic Analyses of Atherosclerotic Vessels From Apolipoprotein E–Deficient Mice Reveal Alterations in Inflammation, Oxidative Stress, and Energy Metabolism. Arterioscler. Thromb. Vasc. Biol. 2005, 25: 2135-2142
[24] Joachim B. R, Yvonne G, Udo R, et al. High-performance anion-exchange chromatography using on-line electrolytic eluent generation for the determination ofmore than 25 intermediates from energy metabolism of mammalian cells in culture. J. Chromatogr. B. 2006, 843: 216-226
[25] S. Lu, X. Sun, C. Shi, et al. Determination of tricarboxylic acid cycle acids and other related substances in cultured mammalian cells by gradient ion-exchange chromatography with suppressed conductibity detection. J. Chromatogr. A. 2003, 1012: 161-168
[26] M.J. Markuszewski, K. Otsuka, S. Terabe, et al. Analysis of carboxylic acid metabolites from the tricarboxylic acid cycle in Bacillus subtils cell extract by capillary electrophroesis using an indirect photometric detection method. J. Chromatogr. A. 2003, 1010: 113-121
[27] James L. Edwards, Robert T. Kennedy, et al. Metabolomic Analysis of Eukaryotic Tissue and Prokaryotes Using Negative Mode MALDI Time-of-Flight Mass Spectrometry. Anal. Chem. 2005, 77: 2201-2209
[28] Leon Coulier, Richard Bas, Sonja Jespersen, et al. Simultaneous Quantitative Analysis of Metabolites Using Ion-Pair Liquid Chromatography-Electrospray Ionization Mass Spectrometry. Anal. Chem. 2006, 78: 6573-6582
[29] B. Luo, K. Groenke, R. Takors, et al. Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography-mass spectrometry. J. Chromatogr. A. 2007, 1147: 153-164
[30]王义明,罗国安.中药复方的化学研究体系.世界科学技术-中药现代化, 1999, 1(1): 16-19
[31]贾延劼,杨于嘉,周燕等.黄芩甙诱导大鼠骨髓基质细胞向神经细胞分化的研究.中华医学杂志. 2002, 82(19): 1337-1341
[32]沈骅睿,吕文科,杨松涛.中药仙茅对骨髓干细胞向神经元细胞定向诱导的实验研究.成都中医药大学学报. 2005, 28(4): 8-11
[33]黎晖,周健洪,陈东风等.龟板对大鼠骨髓间充质干细胞向成骨分化的影响.中药新药与临床药理. 2005, 16(3): 159-161
[34]李瀚旻,晏雪生,罗建君.左归丸药物血清对骨髓间质细胞转化为肝细胞的作用.中国组织工程研究所与临床康复. 2007, 11(28): 5465-5468
[35]孔晓丹,曲鹏,王彦等.三七皂甙诱导骨髓基质细胞分化为心肌样细胞的实验研究.中国心血管病研究杂志. 2005, 3(7): 528-530
[36]汪朝晖,冼绍祥,杨忠奇等.人参总皂甙诱导骨髓间充质干细胞分化为心肌样细胞的实验研究.广州中医药大学学报. 2006, 23(2): 100-103
[37]冼绍祥,杨忠奇,汪朝晖等.黄芪甲苷体外诱导骨髓间充质干细胞分化为心肌样细胞的实验研究.广州中医药大学学报. 2007, 24(1): 37-40
[38]叶能胜,赵艳峰,张荣利等.双龙方干预骨髓间充质干细胞分化过程的蛋白表达.中成药. 2007, 29(1): 24-29
[39]王莎莉,李静,王亚平等.人参皂甙诱导骨髓巨噬细胞表达造血生长因子的研究.解剖学报. 2003, 34(5): 500
[40]陈笛,王莎莉,王亚平等.人参总皂甙诱导人骨髓基质细胞表达GM-MSF的实验研究.重庆医科大学学报. 2003, 28(4): 405
[41]杨莉,蔡卫民.复方丹参滴丸的药理与临床研究.中国医院用药评价与分析, 2005, 5(4): 254
[42]董春桃.丹参的药理作用与临床应用进展.山西医药杂志. 2007, 36(1): 54-55
[43]邓翠娥.丹参的药理作用与临床应用研究进展.时珍国医国药. 2003, 14: 776-777
[44]杜冠华,张均,田同.丹酚酸A对小鼠缺血再灌注致学习记忆功能障碍的改善作用及作用机制.药学学报. 1995, 30(3): 184
[45]王海南.人参皂苷药理研究进展.中国临床药理学与治疗学. 2006, 11(11): 1201-1206
[46]王筠默.人参药理研究的进展.人参研究. 2001, 13(3): 2-10
[47]刘晓晖,朱鲲鹏,许立等.人参皂苷Rb1的研究进展.武警医学院学报. 2006, 15(1): 82-84
[48]李连达,张荣利,刘成源等.双龙方与自体骨髓单个核细胞经心导管移植.中国新药杂志,2003,12(12):999-1004
[49]李连达,张荣利,刘成源等.双龙方对大鼠心肌梗塞的治疗作用.中药新药与临床药理,2004,15(3):149-151
[50]李连达,李贻奎,宁可永等.双龙方对犬心脏血流动力学及心肌耗氧量的影响.中药新药与临床药理,2003,14(6):393-395
[51]罗国安,梁琼麟,刘清飞等.整合化学物质组学的整体系统生物学——中药复方配伍和作用机理研究的整体方法论.世界医学技术—中医药现代化, 2007, 9(1): 10-24
[52]马小琼.人参化学物质基础分析方法研究: [博士学位论文].北京:中国科学院研究生院, 2005
[53] J. Qu, Y. Wang, G. Luo, et al. Vaildated Quantitation of Underivatized Amino Acids in Human Blood Samples by Volatile Ion-Pair Reversed-Phase Liquid Chromatography Coupled to Isotope Dilution Tandem Mass Spectrometry. Anal. Chem. 2002, 74: 2034-2040
[54]余雁.代谢组学中细胞破碎方法的研究.见:中国化学会,编.中国化学会第二十五届学术年会论文摘要集(下册).长春:中国化学会, 2006. 118
[55]叶能胜,张荣利,罗国安等.大鼠骨髓间充质干细胞分化过程的比较蛋白质组学研究.高等学校化学学报, 2006, 27(10):1881-1886
[56] Tomita S, Li R K, Weisel R D, et al. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation 1999, 100: 247-256 Suppl.
[2] Friedenstein A J, Chailakhyan R K and Gerasimov U V. Bone marrow osteogenic stem cells : invitro cultivation and transplantation in diffusion chambers. Cell Tissue Kinet 1987, 20: 263-272
[3] Jiang Y H, Jahagirdar B N, Reinhardt R L, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002, 418: 41-49
[4] Barrias C C, Ribeiro C C, Lamghari M, et al. Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres. J. Biomed. Mater. Res. Part A. 2005, 72A: 57-66
[5] Rickard D J, Sullivan T A, Shenker B J, et al. Induction Of Rapid Osteoblast Differentiation In Rat Bone-Marrow Stromal Cell-Cultures By Dexamethasone And Bmp-2. Dev. Biol. 1994, 161: 218-228
[6] Heino T J, Hentunen T A and Vaananen H K. Conditioned medium from osteocytes stimulates the proliferation of bone marrow mesenchymal stem cells and their differentiation into ostcoblasts. Exp.Cell Res. 2004, 294: 458-468
[7] Martin I, Muraglia A, Campanile G, et al. Fibroblast growth factor-2 supports ex vivo expansion and maintenance of osteogenic precursors from human bone marrow. Endocrinology 1997, 138: 4456-4462
[8] Sugiura F, Kitoh H and Ishiguro N. Osteogenic potential of rat mesenchymal stem cells after several passages. Biochem. Biophys. Res. Commun. 2004, 316: 233-239
[9] Caterson E J, Nesti L J, Li W J, et al. Three-dimensional cartilage formation by bone marrow-derived cells seeded ion polylactide/alginate amalgam. J. Biomed. Mater. Res. 2001, 57: 394-403
[10] Guo X M, Wang C Y, Zhang Y F, et al. Repair of large articular cartilage defects with implants of autologous mesenchymal stem cells seeded into beta-tricalcium phosphate in a sheep model. Tissue Eng. 2004, 10: 1818-1829
[11] Ferrari G, Cusella-De Angelis G, Coletta M, et al. Muscle regeneration by bone marrow derived myogenic progenitors. Science 1998, 279: 1528-1530
[12] Tomita S, Li R K, Weisel R D, et al. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation 1999, 100: 247-256 Suppl
[13] Makino S, Fukuda K, Miyoshi S, et al.Cardiomyocytes can be generated from marrow stromal cells in vitro. J. Clin. Invest. 1999, 103 (5): 697-705
[14] Ye NS (Ye, Neng-Sheng), Chen J (Chen, Jun), Luo GA (Luo, Guo-An), et al. Proteomic profiling of rat bone marrow mesenchymal stem cells induced by 5-azacytidine. Stem Cells and Development, 2006, 15 (5): 665-676
[15] Jeong JA, Lee Y, Lee W, et al. Proteomic analysis of the hydrophobic fraction of mesenchymal stem cells derived from human umbilical cord blood. Molecules and Cells, 2006, 22 (1): 36-43
[16] Wang DJ, Park JS, Chu JSF, et al. Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor beta 1 stimulation. Journal of Biological Chemistry 2004, 279 (42): 43725-43734
[17] Phinney DG, Baddoo M, Dutreil M, et al. Murine mesenchymal stem cells transplanted to the central nervous system of neonatal versus adult mice exhibit distinct engraftment kinetics and express receptors that guide neuronal cell migration. Stem Cells and Development, 2006, 15 (3): 437-447
[18] Forest D, Nishikawa R, Kobayashi H, et al. RNA expression in a cartilaginous fish cell line reveals ancient 3' noncoding regions highly conserved in vertebrates. Proceedings of the National Academy of Sciences of the United States of America, 2007, 104 (4): 1224-1229
[19] Nicholson J K, Connelly J, Lindon J C, et al. Metabonomics:ap latform for studying drug toxicity and gene function. Nat Rev Drug Discov, 2002, 1 (2): 153-161
[20]冉小蓉,梁琼麟,罗国安等.代谢物组学的应用及进展.中成药2005, 27(4): 381-385
[21] Taylor J, King R D, Altmann T, et al. Application of metabolomics to plant genotype discrimination using statistics and machine learning. Bioinform, 2002, 18:S241-S248
[22] Vikram A, Prithiviraj B, Hamzehzarghani H, et al. Valatile metabolite profiling to discriminate diseases of Mc Intosh app le inoculated with fungal pathogens. J Sci FD Agric, 2004, 84(11): 1333-1340
[23] Manuel M, Yuen-Li C, Ursula M, et al. Proteomic and Metabolomic Analyses of Atherosclerotic Vessels From Apolipoprotein E–Deficient Mice Reveal Alterations in Inflammation, Oxidative Stress, and Energy Metabolism. Arterioscler. Thromb. Vasc. Biol. 2005, 25: 2135-2142
[24] Joachim B. R, Yvonne G, Udo R, et al. High-performance anion-exchange chromatography using on-line electrolytic eluent generation for the determination ofmore than 25 intermediates from energy metabolism of mammalian cells in culture. J. Chromatogr. B. 2006, 843: 216-226
[25] S. Lu, X. Sun, C. Shi, et al. Determination of tricarboxylic acid cycle acids and other related substances in cultured mammalian cells by gradient ion-exchange chromatography with suppressed conductibity detection. J. Chromatogr. A. 2003, 1012: 161-168
[26] M.J. Markuszewski, K. Otsuka, S. Terabe, et al. Analysis of carboxylic acid metabolites from the tricarboxylic acid cycle in Bacillus subtils cell extract by capillary electrophroesis using an indirect photometric detection method. J. Chromatogr. A. 2003, 1010: 113-121
[27] James L. Edwards, Robert T. Kennedy, et al. Metabolomic Analysis of Eukaryotic Tissue and Prokaryotes Using Negative Mode MALDI Time-of-Flight Mass Spectrometry. Anal. Chem. 2005, 77: 2201-2209
[28] Leon Coulier, Richard Bas, Sonja Jespersen, et al. Simultaneous Quantitative Analysis of Metabolites Using Ion-Pair Liquid Chromatography-Electrospray Ionization Mass Spectrometry. Anal. Chem. 2006, 78: 6573-6582
[29] B. Luo, K. Groenke, R. Takors, et al. Simultaneous determination of multiple intracellular metabolites in glycolysis, pentose phosphate pathway and tricarboxylic acid cycle by liquid chromatography-mass spectrometry. J. Chromatogr. A. 2007, 1147: 153-164
[30]王义明,罗国安.中药复方的化学研究体系.世界科学技术-中药现代化, 1999, 1(1): 16-19
[31]贾延劼,杨于嘉,周燕等.黄芩甙诱导大鼠骨髓基质细胞向神经细胞分化的研究.中华医学杂志. 2002, 82(19): 1337-1341
[32]沈骅睿,吕文科,杨松涛.中药仙茅对骨髓干细胞向神经元细胞定向诱导的实验研究.成都中医药大学学报. 2005, 28(4): 8-11
[33]黎晖,周健洪,陈东风等.龟板对大鼠骨髓间充质干细胞向成骨分化的影响.中药新药与临床药理. 2005, 16(3): 159-161
[34]李瀚旻,晏雪生,罗建君.左归丸药物血清对骨髓间质细胞转化为肝细胞的作用.中国组织工程研究所与临床康复. 2007, 11(28): 5465-5468
[35]孔晓丹,曲鹏,王彦等.三七皂甙诱导骨髓基质细胞分化为心肌样细胞的实验研究.中国心血管病研究杂志. 2005, 3(7): 528-530
[36]汪朝晖,冼绍祥,杨忠奇等.人参总皂甙诱导骨髓间充质干细胞分化为心肌样细胞的实验研究.广州中医药大学学报. 2006, 23(2): 100-103
[37]冼绍祥,杨忠奇,汪朝晖等.黄芪甲苷体外诱导骨髓间充质干细胞分化为心肌样细胞的实验研究.广州中医药大学学报. 2007, 24(1): 37-40
[38]叶能胜,赵艳峰,张荣利等.双龙方干预骨髓间充质干细胞分化过程的蛋白表达.中成药. 2007, 29(1): 24-29
[39]王莎莉,李静,王亚平等.人参皂甙诱导骨髓巨噬细胞表达造血生长因子的研究.解剖学报. 2003, 34(5): 500
[40]陈笛,王莎莉,王亚平等.人参总皂甙诱导人骨髓基质细胞表达GM-MSF的实验研究.重庆医科大学学报. 2003, 28(4): 405
[41]杨莉,蔡卫民.复方丹参滴丸的药理与临床研究.中国医院用药评价与分析, 2005, 5(4): 254
[42]董春桃.丹参的药理作用与临床应用进展.山西医药杂志. 2007, 36(1): 54-55
[43]邓翠娥.丹参的药理作用与临床应用研究进展.时珍国医国药. 2003, 14: 776-777
[44]杜冠华,张均,田同.丹酚酸A对小鼠缺血再灌注致学习记忆功能障碍的改善作用及作用机制.药学学报. 1995, 30(3): 184
[45]王海南.人参皂苷药理研究进展.中国临床药理学与治疗学. 2006, 11(11): 1201-1206
[46]王筠默.人参药理研究的进展.人参研究. 2001, 13(3): 2-10
[47]刘晓晖,朱鲲鹏,许立等.人参皂苷Rb1的研究进展.武警医学院学报. 2006, 15(1): 82-84
[48]李连达,张荣利,刘成源等.双龙方与自体骨髓单个核细胞经心导管移植.中国新药杂志,2003,12(12):999-1004
[49]李连达,张荣利,刘成源等.双龙方对大鼠心肌梗塞的治疗作用.中药新药与临床药理,2004,15(3):149-151
[50]李连达,李贻奎,宁可永等.双龙方对犬心脏血流动力学及心肌耗氧量的影响.中药新药与临床药理,2003,14(6):393-395
[51]罗国安,梁琼麟,刘清飞等.整合化学物质组学的整体系统生物学——中药复方配伍和作用机理研究的整体方法论.世界医学技术—中医药现代化, 2007, 9(1): 10-24
[52]马小琼.人参化学物质基础分析方法研究: [博士学位论文].北京:中国科学院研究生院, 2005
[53] J. Qu, Y. Wang, G. Luo, et al. Vaildated Quantitation of Underivatized Amino Acids in Human Blood Samples by Volatile Ion-Pair Reversed-Phase Liquid Chromatography Coupled to Isotope Dilution Tandem Mass Spectrometry. Anal. Chem. 2002, 74: 2034-2040
[54]余雁.代谢组学中细胞破碎方法的研究.见:中国化学会,编.中国化学会第二十五届学术年会论文摘要集(下册).长春:中国化学会, 2006. 118
[55]叶能胜,张荣利,罗国安等.大鼠骨髓间充质干细胞分化过程的比较蛋白质组学研究.高等学校化学学报, 2006, 27(10):1881-1886
[56] Tomita S, Li R K, Weisel R D, et al. Autologous transplantation of bone marrow cells improves damaged heart function. Circulation 1999, 100: 247-256 Suppl.