实验兔血管内膜损伤后不同阶段PDGF-B链、C-myc基因表达及中药对其拮抗作用机制研究
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摘要
目的：依据中医中药理论在继承的基础上，借鉴分子生物学方法，探讨实验性兔胸主动脉内膜损伤后，血管内膜增殖早期主要调控因子-血小板源生长因子(PDGF-B)及相关原癌基因C-myc的病理机制。通过芪麦通对PDGF-B以及C-myc拮抗作用，从不同层次阐明中药益气活血化瘀能在细胞和基因水平发挥干预作用，最终为达到在内膜损伤早期防治血管再狭窄提供新的实验依据。
方法：模拟PTCA术后再狭窄的模型，采用球囊导管剥脱术，造成实验兔胸主动脉内膜损伤，分别于内膜损伤后第1周、第2周、第4周处死，取兔胸主动脉组织，通过HE染色观察血管大体形态学变化；用原位杂交法观察测定PDGF-B及C-myc在血管损伤部位不同阶段的动态表达；采用逆转录PCR(RT-PCR)检测PDGF-BmRNA转录；Westem免疫印迹方法检测PDGF-B蛋白质表达水平。
结果：显微镜下观察组织形态学变化：HE染色可见损伤后造成内皮及部分内膜坏死脱落、内弹力板断裂、血栓形成。损伤后1周，内膜增厚并主要由细胞构成，大多为SMCa-actin阳性，偶见泡沫细胞、淋巴细胞、少许中性细胞。数周后靠近腔内的细胞仍保持增生肥大，但用药组损伤后一周内膜阳性细胞分布明显减少，中层可见少数散在的弱阳性细胞，数周后仍保持稳定的水平。
原位杂交镜下观察测定：PDGF-B链在血管损伤部位不同阶段的动态表达，以及中药对它们的影响。正常的血管内膜可见少量阳性细胞表达，损伤后1周内膜阳性细胞分布明显增多，中膜亦可见少量的散在的阳性细胞分布，4周后呈稳定的水平下降。但用药组损伤1周后，内膜阳性细胞分布明显减少，中层可见少数散在的弱阳性细胞，数周后仍保持稳定的水平。并且中药组与西药组比较阳性表达减弱更为明显。P＜0.001
逆转录(RT-PCR)方法检测：显示胸主动脉血管内膜损伤后、第一周对照组明显高于正常组、P＜0.001；第四周与前两周比逐渐恢复。结果显示胸主动脉内膜损伤后早期PDGF-BmRNA呈增高趋势。而中药芪麦通对胸主动脉血管内膜损伤早期PDGF-BmRNA转录有明显拮抗作用。
Westem免疫印迹方法检测：显示胸主动脉血管内膜损伤后、第一周对照组明显高于正常组、P＜0.001；第四周与前两周比蛋白表达逐渐减少。结果显示胸主动脉内膜损伤后早期PDGF-B蛋白呈增高趋势，而中药芪麦通对胸主动脉血管内膜损伤早期PDGF-B蛋白的表达有明显拮抗作用。
结论：在损伤早期被激活后血小板产生，血小板源生长因子PDGF，被趋势化了的内皮细胞、巨噬细胞，在损伤部位聚集，诱生PDGF－B在早期出现一个高峰，而这些生长因子通过自分泌和旁分泌途径启动血管平滑肌的增殖，这同中医的气虚血瘀状态与病理产物积聚有关。由于病理代谢产物刺激基因异常表达，诱导原癌基因表达并进而分泌某些细胞或体液因子，引起组织异常增生导致再狭窄的发生而在整个病变的发展过程中由于气虚血瘀贯穿本病始终。而芪麦通对其有明显的拮抗作用。说明益气活血化瘀药能在细胞和基因水平发挥干预作用。即使受创血管内皮细胞修复再生同时，芪麦通对其有明显的拮抗作用。又可以迅速抑制受创血管周围血小板的聚集及炎性渗出行血中之气功。气血同治，气阴双补而不滋腻恋邪，既可补虚固本亦可以活血化瘀，从药物组成配伍合用可体现气虚瘀互补功效。
PurposerThe experiment was based on the Traditional Chinese Medicine theory and used the method of molecular biology.The purpose of this experiment is to study the mechanism of PDGF-B which is the major regulation and control factor during earlier period of proliferation of post-injuried chest thoracic aorta of the rabbit. Traditional Chinese Medicine QMT can prevent: and intervene in the role of PDGF-B both in cell and gene levels.This experiment also offer the evidence on QMThow to prevent and cure the blood vessel restosis after earlier period of tunica intima damaged.
Method :Foley's tube denudation was used to cause the chest thoracic aorta of the rabbit damaged.The animals were killed after tunica intima damaged lw.2w,4w ,then extracted the chest thoracic aorta constitution of the rabbit..The changes of blood vessel morphology were observed by HE staining. The expression of PDGF-B at different stage was assayed in situ hybridization.The transcription and protein expression of PDGF-B mRNA were detected by the methods of RT-PCR and western immunoblot.
Result: The protein expression of PDGF-B was phancro-reinforcement after earlier period(lw-2w) of tunica intima damaged.(P<0.01)During 3w and 4w the expression of PDGF-B was reduced gradually.HE staining manifested that the mecrosis and abscissiou of endothelium and demi-endometium were caused by injurying.The inter-elasticity blade broke and thrombo formatted.Afer inuried one week,the endometrium thickened and consisted of cells.Most of them is SMCa-actin male.The foam cell,lymphocyte and a little reutrality can be seen occasionally.The cells near to the intraluminal still keep hyperplasia hypertrophy after several weeks.The spread of indometrium male cells were reduced clearly in one week of medication group.A little feeble male cells were seen in uveal tract and still keep nesistant level after several weeks. In situ hybridization.assay: PDGF-B can express at different stage. The expression of a litter male cells can be seen in the normal intima. The spread of the male cells in intima were increased clear
ly after one week of injury .A little feeble male cells were seen in uveal tract and still keep nesistant level after several weeks. The expression of the TCM group were more weakened than western group.
RT-PCR manifest that the expression of the control group was more than normal group at first week after chest thoracic aorta injuried. Compared the 4l week with the first two weeks, the expression was recovered gradually. It indicated MRNA had the growing tendency during the earlier period of tunica intima damaged. While QMTcould prevent the transcription of MRAN in earlier period of chest tunica intima damaged.
Western immunoblot assay: The expression of PDGF-B in the control group was more than normal group at first week after chest thoracic aorta injuried. Compared the 4th week with the first two weeks, the expression was reduced clearly.
Conclusion: PDGF-B is the major regulation an control factor during the earlier period of proliferation of tunica intima. QMT can prevent the rote of PDGF-B. we can deduce that the blood platelet is activated during the earlier period of injury. PDGF-B is collected in the injuried locus by
endothelial cell and histocyte to induce the PDGF-B to produce a pinnacle at earlier period. These grouth factors start the angio-blood muscle proliferation through autocrine and paracrine, These are related to the blood stagnant of TCM. Metabolite simulates gene abnomal expression and induces protooncogene expression, then excrete some cells and body fluide and cause constition abnormally hyperplasia and restenosis. Reinforcing QI actirating the blood and eliminating the stagnant QMTcan have intervene role both in cell and gene level. It can pierosis and regenerate the damaged endotheliurn, at the same time it can prevent the platelet around and inflammation bleeding damaged vessel collecting. QMT can be simultaneous treatment of QI and blood, and reinforcing. Both Qi and Yin. It can not only reinforcing deficiency and tre
方法：模拟PTCA术后再狭窄的模型，采用球囊导管剥脱术，造成实验兔胸主动脉内膜损伤，分别于内膜损伤后第1周、第2周、第4周处死，取兔胸主动脉组织，通过HE染色观察血管大体形态学变化；用原位杂交法观察测定PDGF-B及C-myc在血管损伤部位不同阶段的动态表达；采用逆转录PCR(RT-PCR)检测PDGF-BmRNA转录；Westem免疫印迹方法检测PDGF-B蛋白质表达水平。
结果：显微镜下观察组织形态学变化：HE染色可见损伤后造成内皮及部分内膜坏死脱落、内弹力板断裂、血栓形成。损伤后1周，内膜增厚并主要由细胞构成，大多为SMCa-actin阳性，偶见泡沫细胞、淋巴细胞、少许中性细胞。数周后靠近腔内的细胞仍保持增生肥大，但用药组损伤后一周内膜阳性细胞分布明显减少，中层可见少数散在的弱阳性细胞，数周后仍保持稳定的水平。
原位杂交镜下观察测定：PDGF-B链在血管损伤部位不同阶段的动态表达，以及中药对它们的影响。正常的血管内膜可见少量阳性细胞表达，损伤后1周内膜阳性细胞分布明显增多，中膜亦可见少量的散在的阳性细胞分布，4周后呈稳定的水平下降。但用药组损伤1周后，内膜阳性细胞分布明显减少，中层可见少数散在的弱阳性细胞，数周后仍保持稳定的水平。并且中药组与西药组比较阳性表达减弱更为明显。P＜0.001
逆转录(RT-PCR)方法检测：显示胸主动脉血管内膜损伤后、第一周对照组明显高于正常组、P＜0.001；第四周与前两周比逐渐恢复。结果显示胸主动脉内膜损伤后早期PDGF-BmRNA呈增高趋势。而中药芪麦通对胸主动脉血管内膜损伤早期PDGF-BmRNA转录有明显拮抗作用。
Westem免疫印迹方法检测：显示胸主动脉血管内膜损伤后、第一周对照组明显高于正常组、P＜0.001；第四周与前两周比蛋白表达逐渐减少。结果显示胸主动脉内膜损伤后早期PDGF-B蛋白呈增高趋势，而中药芪麦通对胸主动脉血管内膜损伤早期PDGF-B蛋白的表达有明显拮抗作用。
结论：在损伤早期被激活后血小板产生，血小板源生长因子PDGF，被趋势化了的内皮细胞、巨噬细胞，在损伤部位聚集，诱生PDGF－B在早期出现一个高峰，而这些生长因子通过自分泌和旁分泌途径启动血管平滑肌的增殖，这同中医的气虚血瘀状态与病理产物积聚有关。由于病理代谢产物刺激基因异常表达，诱导原癌基因表达并进而分泌某些细胞或体液因子，引起组织异常增生导致再狭窄的发生而在整个病变的发展过程中由于气虚血瘀贯穿本病始终。而芪麦通对其有明显的拮抗作用。说明益气活血化瘀药能在细胞和基因水平发挥干预作用。即使受创血管内皮细胞修复再生同时，芪麦通对其有明显的拮抗作用。又可以迅速抑制受创血管周围血小板的聚集及炎性渗出行血中之气功。气血同治，气阴双补而不滋腻恋邪，既可补虚固本亦可以活血化瘀，从药物组成配伍合用可体现气虚瘀互补功效。
PurposerThe experiment was based on the Traditional Chinese Medicine theory and used the method of molecular biology.The purpose of this experiment is to study the mechanism of PDGF-B which is the major regulation and control factor during earlier period of proliferation of post-injuried chest thoracic aorta of the rabbit. Traditional Chinese Medicine QMT can prevent: and intervene in the role of PDGF-B both in cell and gene levels.This experiment also offer the evidence on QMThow to prevent and cure the blood vessel restosis after earlier period of tunica intima damaged.
Method :Foley's tube denudation was used to cause the chest thoracic aorta of the rabbit damaged.The animals were killed after tunica intima damaged lw.2w,4w ,then extracted the chest thoracic aorta constitution of the rabbit..The changes of blood vessel morphology were observed by HE staining. The expression of PDGF-B at different stage was assayed in situ hybridization.The transcription and protein expression of PDGF-B mRNA were detected by the methods of RT-PCR and western immunoblot.
Result: The protein expression of PDGF-B was phancro-reinforcement after earlier period(lw-2w) of tunica intima damaged.(P<0.01)During 3w and 4w the expression of PDGF-B was reduced gradually.HE staining manifested that the mecrosis and abscissiou of endothelium and demi-endometium were caused by injurying.The inter-elasticity blade broke and thrombo formatted.Afer inuried one week,the endometrium thickened and consisted of cells.Most of them is SMCa-actin male.The foam cell,lymphocyte and a little reutrality can be seen occasionally.The cells near to the intraluminal still keep hyperplasia hypertrophy after several weeks.The spread of indometrium male cells were reduced clearly in one week of medication group.A little feeble male cells were seen in uveal tract and still keep nesistant level after several weeks. In situ hybridization.assay: PDGF-B can express at different stage. The expression of a litter male cells can be seen in the normal intima. The spread of the male cells in intima were increased clear
ly after one week of injury .A little feeble male cells were seen in uveal tract and still keep nesistant level after several weeks. The expression of the TCM group were more weakened than western group.
RT-PCR manifest that the expression of the control group was more than normal group at first week after chest thoracic aorta injuried. Compared the 4l week with the first two weeks, the expression was recovered gradually. It indicated MRNA had the growing tendency during the earlier period of tunica intima damaged. While QMTcould prevent the transcription of MRAN in earlier period of chest tunica intima damaged.
Western immunoblot assay: The expression of PDGF-B in the control group was more than normal group at first week after chest thoracic aorta injuried. Compared the 4th week with the first two weeks, the expression was reduced clearly.
Conclusion: PDGF-B is the major regulation an control factor during the earlier period of proliferation of tunica intima. QMT can prevent the rote of PDGF-B. we can deduce that the blood platelet is activated during the earlier period of injury. PDGF-B is collected in the injuried locus by
endothelial cell and histocyte to induce the PDGF-B to produce a pinnacle at earlier period. These grouth factors start the angio-blood muscle proliferation through autocrine and paracrine, These are related to the blood stagnant of TCM. Metabolite simulates gene abnomal expression and induces protooncogene expression, then excrete some cells and body fluide and cause constition abnormally hyperplasia and restenosis. Reinforcing QI actirating the blood and eliminating the stagnant QMTcan have intervene role both in cell and gene level. It can pierosis and regenerate the damaged endotheliurn, at the same time it can prevent the platelet around and inflammation bleeding damaged vessel collecting. QMT can be simultaneous treatment of QI and blood, and reinforcing. Both Qi and Yin. It can not only reinforcing deficiency and tre
引文
[1] 《内经》
[2] 张仲景《伤寒杂病论》 东汉
[3] 唐宗海《血证论．瘀血》 清代
[4] 《医林改错》王清任 清代
[5] 《黄帝内经研究大成》，北京出版社，1997，1137—1138页
[6] 毛以林 袁肇凯冠心病血瘀证证候本质研究最新进展 中国中医药信息杂志 2002
[7] 冼绍祥 黄鹂等益气、行气活血法在冠心病心血瘀阻证中的作用机理探讨新中医 2002
[8] 程文立 乔占兵等加味大黄蛰虫胶囊对ADP诱导大鼠血小板聚集的影响 河南中医 2002
[9] 李朝敏 丁红原发性高血压患者红细胞流变学异常与血瘀证关系临床研究中国医药学报 2002
[10] 李朝敏 丁红心元胶囊治疗胸痹心肾阴虚兼心血瘀阴证临床研究中药药理与临床 2002
[11] 石云李新等活血化瘀法治疗老年病的体会中国中医药信息杂志 2002
[12] 郭键为血瘀证证候本质活血法作用机理探讨陕西中医学院学报 2002
[13] Bareff TB benditt EP Sis gene transcript levels are elevated in human after osclerotic lesions compared to normal artery proc natl Acad SciUSA1987 84:1099
[14] Barrett TB, Benditt EP. Platelet-derived gowth factor gene expression in human atherosclerosis plaque and nonmal artery wall. Proc Natl Acad Sci USA, 1988,85:2810
[15] Gordon AA, Fems FW, Katherine H, et al. Inluibition of nrointimal smooth muscle accumulation after angioplastu by an anti body to PDGF. Science, 1991,25(3):1129
[16] linderv, reidy m, apdgf ligand and neceptor expression by lang vessel endothium in vivo [j]Amj p pathol1995.146(6)1488-1497
[17] lindner vVC Gachelli CM ,Schvartz. SM et lation of snooth muscle cells in injured rat a platelet-derive growth factor-B chain m res 1995 76(6)951-957
[18] joseph PM Thompson BT, et al effect of hypoxis on the secretion of platelet-derived growfictor by bovine pulmonary artery endothelial cell AmJ Respir Dis1990;141:A346
[19] GiJun ,Si fusheng Interferon Y inhibits expression of PDGF-β mRNA by smooth muscle cells in injured rabbit trarsluminal balloon angioplasty Chinese Medicail] Joumal 2001;114(2):214-223
[20] Khichigian LM, williams AJCollins T Interplay of sP-I and Egr-I in the prosximal PDGF-A promoter in cultured vascular endothelial cells J Biol chem 1994,269;28648
[21] Khichigian LM fries JWU Bez Mw et al Novel cis-acting elements in the human platelet-derived growth factor B-chain cone promoter that mediated gene expression in chltured vascular endothelial cells J Biol Chem, 1994-269:27679
[22] Shen Chuanl '-Lu the regulation of PDGF-Bchain gene expression and its
significance in common cardiovascular diseases Chinese Joumal of pathophysiology 1999, 15(12) 1138-1141
[23] 王关嵩 钱桂生等血小板源生长因子对血管平滑肌细胞的作用研究国外医学临床生物学与检验学分册 1997，18(4)149-151
[24] Nimni ME palypeptide growth factors targeted delivery ystems. Biomaterials 1997;18(18) 1 ,201
[25] Relly JF Kumari VG Alterations in fibroblast growth factor receptor expression following brain ingury Exp Neurol 1996:140(2) 139
[26] Brehm BRWolfSCBertsch Det al Effectl of nebivolol on proliferation and apoptosis of human coronary artery smooth muscle and endothelial cells [J]Cardiovasc Res, 2001,49(2):430-439
[27] Okam oto Smassak ilnden Setsuda M et al Effects of trapidil PDGF antagonist in preventing restenosis after percotaneous translum inalcoronary angioplasty [J] Am Heart J 1992,123(6) 1439-1444
[28] myers PRTanner MAVascular end regulation of extracellular matrix colla nitric oxide Arterioscler Thromb Vasc 1998 18(5)717-722
[29] 向定成，黄大显等血小板源生长因子与冠状动脉成形术后再狭窄 国外医学生理，病理科学与临床分册再狭窄
[30] Hoberg E, Dietz R, Frees U et al. Veraparmil treatment after cornary angioplasty in patients at high risk of recurrent sternosis. Haert J, 1994:71:254
[31] Setsuda M, Inden M, Hiraoks N et al. Probucol therapy in the prvention of restenosis after successful PTCA. Clin Ther, 1993:15374
[32] Hirayama A, Nanto S, Ohara T et al. Preventive effect on restensis after PTCA by ebselen:A newly sunthesized antiinflammatoragent. J Am Coll Cardiol, 1992;19:259A
[33] Robert J. Griffin, Brent W. Williams, Robert Wild, Julie M. Cherrington, Heonjoo Park, and Chang W. Song Simultaneous Inhibition of the Receptor Kinase Activity of Vascular Endothelial, Fibroblast, and Platelet-derived Growth Factors Suppresses Tumor Growth and Enhances Tumor Radiation Response Cancer research2002,62(6).-1702-170
[34] Kristian Pietras, Kristofer Rubin, Elisabeth Buchdunger, Inhibition of PDGF Receptor Signaling in Tumor Stroma Enhances Antitumor Effect of Chemotherapy Cancer research 2002,62(19).-5476-54
[35] Mohamed E. Abdel-Wanis and Norio Kawahara The role of neurofibromin and melatonin in pathogenesis of pseudarthrosis after spinal fusion for neurofibromatous scoliosis Medical Hypotheses2002,58(5).-395-398
[36] Agapi Kataki, Philippe Scheid, Maud Piet, Béatrice Marie, Nadine Martinet, Yves Martinet Tumor infiltrating iymphocytes and macrophages have a potential dual role in lung cancer by supporting both host-defense and tumor progression Journal of Laboratory & Clinical Medicine 2002,140(5).-320-328
[37] T. Kohyama, X. Liu, F. Q. Wen, H. J. Kim, H. Takizawa, S. I. Rennard
Potentiation of human lung fibroblast chemotaxis by the thromboxane A 2 analog U-46619 Journal of Laboratory & Clinical Medicine -2002,139(1).-43-49
[38] Tadashi Kohyama, Xiangde Liu, Fu-Qiang Wen, Nerve growth factor stimulates fibronectin-induced fibroblast migration Journal of Laboratory & Clinical Medicine 2002, 140 (5). -329-33
[39] S. Goodall, K. E. Porter Aorti Enhanced Invasive Properties Exhibited by Smooth Muscle Cells are Associated with Elevated Production of MMP-2 in Patients with c Aneurysms European Journal of Vascular and Endovascular Surgery 002, 24 (1). -72-8
[40] Linnea M. Baudhuin, Kelly L. Cristina Akt Activation Induced by Lysophosphatidic Acid and Sphingosine-1-phosphate Requires Both Mitogen-Activated Protein Kinase Kinase and p38 Mitogen-Activated Protein Kinase and Is Cell-Line-Specific Molecular Pharmacology 2002,62(3).-660-
[41] 王明艳 周坤福 徐力 分子生物学与中医药研究 上海中医药大学出版社[43]
[42] 王银叶 王景燕付逸龙；L-精氨酸L-门冬氨酸盐对血小板功能的抑制生理学报2001年第4期第53卷
[43] 兰建平 朱建华等血管内局部给药的实验研究中国循环杂志 1999，14；81
[44] 吴明医学科学六个新课题科学时报2002-04-12
[45] 陈镜合现代中医之路广州中医药大学学报2001-9
[2] 张仲景《伤寒杂病论》 东汉
[3] 唐宗海《血证论．瘀血》 清代
[4] 《医林改错》王清任 清代
[5] 《黄帝内经研究大成》，北京出版社，1997，1137—1138页
[6] 毛以林 袁肇凯冠心病血瘀证证候本质研究最新进展 中国中医药信息杂志 2002
[7] 冼绍祥 黄鹂等益气、行气活血法在冠心病心血瘀阻证中的作用机理探讨新中医 2002
[8] 程文立 乔占兵等加味大黄蛰虫胶囊对ADP诱导大鼠血小板聚集的影响 河南中医 2002
[9] 李朝敏 丁红原发性高血压患者红细胞流变学异常与血瘀证关系临床研究中国医药学报 2002
[10] 李朝敏 丁红心元胶囊治疗胸痹心肾阴虚兼心血瘀阴证临床研究中药药理与临床 2002
[11] 石云李新等活血化瘀法治疗老年病的体会中国中医药信息杂志 2002
[12] 郭键为血瘀证证候本质活血法作用机理探讨陕西中医学院学报 2002
[13] Bareff TB benditt EP Sis gene transcript levels are elevated in human after osclerotic lesions compared to normal artery proc natl Acad SciUSA1987 84:1099
[14] Barrett TB, Benditt EP. Platelet-derived gowth factor gene expression in human atherosclerosis plaque and nonmal artery wall. Proc Natl Acad Sci USA, 1988,85:2810
[15] Gordon AA, Fems FW, Katherine H, et al. Inluibition of nrointimal smooth muscle accumulation after angioplastu by an anti body to PDGF. Science, 1991,25(3):1129
[16] linderv, reidy m, apdgf ligand and neceptor expression by lang vessel endothium in vivo [j]Amj p pathol1995.146(6)1488-1497
[17] lindner vVC Gachelli CM ,Schvartz. SM et lation of snooth muscle cells in injured rat a platelet-derive growth factor-B chain m res 1995 76(6)951-957
[18] joseph PM Thompson BT, et al effect of hypoxis on the secretion of platelet-derived growfictor by bovine pulmonary artery endothelial cell AmJ Respir Dis1990;141:A346
[19] GiJun ,Si fusheng Interferon Y inhibits expression of PDGF-β mRNA by smooth muscle cells in injured rabbit trarsluminal balloon angioplasty Chinese Medicail] Joumal 2001;114(2):214-223
[20] Khichigian LM, williams AJCollins T Interplay of sP-I and Egr-I in the prosximal PDGF-A promoter in cultured vascular endothelial cells J Biol chem 1994,269;28648
[21] Khichigian LM fries JWU Bez Mw et al Novel cis-acting elements in the human platelet-derived growth factor B-chain cone promoter that mediated gene expression in chltured vascular endothelial cells J Biol Chem, 1994-269:27679
[22] Shen Chuanl '-Lu the regulation of PDGF-Bchain gene expression and its
significance in common cardiovascular diseases Chinese Joumal of pathophysiology 1999, 15(12) 1138-1141
[23] 王关嵩 钱桂生等血小板源生长因子对血管平滑肌细胞的作用研究国外医学临床生物学与检验学分册 1997，18(4)149-151
[24] Nimni ME palypeptide growth factors targeted delivery ystems. Biomaterials 1997;18(18) 1 ,201
[25] Relly JF Kumari VG Alterations in fibroblast growth factor receptor expression following brain ingury Exp Neurol 1996:140(2) 139
[26] Brehm BRWolfSCBertsch Det al Effectl of nebivolol on proliferation and apoptosis of human coronary artery smooth muscle and endothelial cells [J]Cardiovasc Res, 2001,49(2):430-439
[27] Okam oto Smassak ilnden Setsuda M et al Effects of trapidil PDGF antagonist in preventing restenosis after percotaneous translum inalcoronary angioplasty [J] Am Heart J 1992,123(6) 1439-1444
[28] myers PRTanner MAVascular end regulation of extracellular matrix colla nitric oxide Arterioscler Thromb Vasc 1998 18(5)717-722
[29] 向定成，黄大显等血小板源生长因子与冠状动脉成形术后再狭窄 国外医学生理，病理科学与临床分册再狭窄
[30] Hoberg E, Dietz R, Frees U et al. Veraparmil treatment after cornary angioplasty in patients at high risk of recurrent sternosis. Haert J, 1994:71:254
[31] Setsuda M, Inden M, Hiraoks N et al. Probucol therapy in the prvention of restenosis after successful PTCA. Clin Ther, 1993:15374
[32] Hirayama A, Nanto S, Ohara T et al. Preventive effect on restensis after PTCA by ebselen:A newly sunthesized antiinflammatoragent. J Am Coll Cardiol, 1992;19:259A
[33] Robert J. Griffin, Brent W. Williams, Robert Wild, Julie M. Cherrington, Heonjoo Park, and Chang W. Song Simultaneous Inhibition of the Receptor Kinase Activity of Vascular Endothelial, Fibroblast, and Platelet-derived Growth Factors Suppresses Tumor Growth and Enhances Tumor Radiation Response Cancer research2002,62(6).-1702-170
[34] Kristian Pietras, Kristofer Rubin, Elisabeth Buchdunger, Inhibition of PDGF Receptor Signaling in Tumor Stroma Enhances Antitumor Effect of Chemotherapy Cancer research 2002,62(19).-5476-54
[35] Mohamed E. Abdel-Wanis and Norio Kawahara The role of neurofibromin and melatonin in pathogenesis of pseudarthrosis after spinal fusion for neurofibromatous scoliosis Medical Hypotheses2002,58(5).-395-398
[36] Agapi Kataki, Philippe Scheid, Maud Piet, Béatrice Marie, Nadine Martinet, Yves Martinet Tumor infiltrating iymphocytes and macrophages have a potential dual role in lung cancer by supporting both host-defense and tumor progression Journal of Laboratory & Clinical Medicine 2002,140(5).-320-328
[37] T. Kohyama, X. Liu, F. Q. Wen, H. J. Kim, H. Takizawa, S. I. Rennard
Potentiation of human lung fibroblast chemotaxis by the thromboxane A 2 analog U-46619 Journal of Laboratory & Clinical Medicine -2002,139(1).-43-49
[38] Tadashi Kohyama, Xiangde Liu, Fu-Qiang Wen, Nerve growth factor stimulates fibronectin-induced fibroblast migration Journal of Laboratory & Clinical Medicine 2002, 140 (5). -329-33
[39] S. Goodall, K. E. Porter Aorti Enhanced Invasive Properties Exhibited by Smooth Muscle Cells are Associated with Elevated Production of MMP-2 in Patients with c Aneurysms European Journal of Vascular and Endovascular Surgery 002, 24 (1). -72-8
[40] Linnea M. Baudhuin, Kelly L. Cristina Akt Activation Induced by Lysophosphatidic Acid and Sphingosine-1-phosphate Requires Both Mitogen-Activated Protein Kinase Kinase and p38 Mitogen-Activated Protein Kinase and Is Cell-Line-Specific Molecular Pharmacology 2002,62(3).-660-
[41] 王明艳 周坤福 徐力 分子生物学与中医药研究 上海中医药大学出版社[43]
[42] 王银叶 王景燕付逸龙；L-精氨酸L-门冬氨酸盐对血小板功能的抑制生理学报2001年第4期第53卷
[43] 兰建平 朱建华等血管内局部给药的实验研究中国循环杂志 1999，14；81
[44] 吴明医学科学六个新课题科学时报2002-04-12
[45] 陈镜合现代中医之路广州中医药大学学报2001-9