同种异体骨髓间充质干细胞穴位注射零天与七天治疗急性心肌梗死兔的对比研究
摘要
目的:探讨兔骨髓间充质干细胞(MSCs)穴位注射对缺血心肌细胞的修复重建能力,同时进行兔骨髓间充质干细胞(MSCs)穴位注射零天与七天对梗死区心肌细胞凋亡的对比研究。采用脱氧核苷酸末端转移酶介导原位缺口标记(TUNEL法)确定细胞凋亡数,免疫组化SABC方法及RT-PCR和蛋白印迹法观察心肌细胞中Fas、FasL、Bcl-2蛋白及mRNA的表达。
方法:
1.将已建立心肌梗死模型的日本大耳白兔随机分为3组,模型组、穴位注射干细胞零天组、穴位注射干细胞七天组。用开胸结扎冠状动脉左室支的方法建立急性心肌梗死模型,对梗死后的兔分两组分别在零天与七天穴位注射相同数量的已经分离、培养、扩增、标记后的MSCs悬液。在梗死后五周分别对各组兔心肌梗死区及周边区进行Brdu免疫组化染色以观察穴位注射MSCs是否迁移至梗死区,对Brdu染色阳性的细胞复染心肌特异性肌钙蛋白T染色以观察迁移的细胞是否分化及分化的性质。
2.采用脱氧核苷酸末端转移酶介导原位缺口标记(TUNEL法)确定细胞凋亡数,免疫组化SABC方法及RT-PCR和蛋白印迹法观察心肌细胞中Fas、FasL、Bcl-2蛋白及mRNA的表达。
结果:
1.穴位注射MSCs各组梗死区组织免疫组化Brdu染色阳性;心肌特异性肌钙蛋白T染色阳性。
2.免疫组化示:梗死区凋亡细胞的表达,模型组高于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组(P＜0.05),穴位注射干细胞七天组低于穴位注射干细胞零天组,但无统计学差异(P＞0.05);Fas蛋白的表达,模型组高于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组,穴位注射干细胞七天组低于穴位注射干细胞零天组(P＜0.05);FasL蛋白的表达,穴位注射干细胞零天组高于正常组、模型组、穴位注射干细胞七天组(P＜0.05);Bcl-2蛋白的表达,模型组低于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组(P＜0.05),穴位注射干细胞七天组高于穴位注射干细胞零天组,但无统计学差异(P＞0.05)。
3.RT-PCR与Western Blot显示:与正常组比较,Fas mRNA水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);FasL mRNA水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);Bcl-2 mRNA水平:模型组明显下降(P＜0.05),穴位注射干细胞的两组有所升高,但升高程度不同,穴位注射零天组明显升高(P＜0.05),穴位注射七天组虽有升高但无统计学意义(P＞0.05)。与模型组比较,Fas mRNA水平:穴位注射干细胞的两组明显下降(P＜0.05);FasL mRNA水平:穴位注射干细胞的两组明显下降(P＜0.01);Bcl-2 mRNA:穴位注射干细胞的两组明显升高(P＜0.05)。与穴位注射干细胞零天比较,Fas mRNA水平:穴位注射干细胞七天组明显下降(P＜0.05);FASL mRNA水平:穴位注射干细胞七天组明显下降(P＜0.05);Bcl-2 mRNA水平:穴位注射干细胞七天组也明显下降(P＜0.05)。与正常组比较,Fas与FasL蛋白水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);Bcl-2蛋白水平:模型组明显下降(P＜0.05),穴位注射干细胞的两组有所升高,但升高程度不同,穴位注射零天组明显升高(P＜0.05),穴位注射七天组明显升高(P＜0.05)。与模型组比较,Fas蛋白:穴位注射干细胞的两组明显下降(P＜0.05);FasL蛋白:穴位注射干细胞的两组明显下降(P＜0.05);Bcl-2蛋白:穴位注射干细胞的两组明显升高(P＜0.05)。与穴位注射干细胞零天比较,Fas蛋白:穴位注射干细胞七天组明显下降(P＜0.05);FASL蛋白:穴位注射干细胞七天组明显下降(P＜0.05);Bcl-2蛋白:穴位注射干细胞七天组也明显下降(P＜0.05)。
结论:
1.穴位注射兔骨髓间充质干细胞可迁移至梗死区,并可以在梗死区分化为类心肌样细胞。
2.穴位注射骨髓间充质干细胞可使梗死及周围区心肌细胞凋亡数减少,其机制可能是通过Fas、FasL蛋白的减少、Bcl-2蛋白的增多来调节的。
3.穴位注射骨髓间充质干细胞七天较穴位注射骨髓间充质干细胞零天更能减少凋亡细胞的数量。
Objective: To investigate the reconstruction capacity of rabbit bone marrow-derived mesenchymal stem cells (MSCs) Point-cell transplantation for ischemic myocyte, at the same time to make a comparative research on cardiomyocyte apoptosis in infarct area between rabbit bone marrow mesenchymal stem cells (MSCs) transplantation Point of zero-day and seven days. To determine the number of cell apoptosis by TUNEL, and to observe the expression of Fas, FasL, Bcl-2 protein and mRNA in myocardial cells by immunohistochemistry SABC methods, RT-PCR and Western blot.
Methods: (1) The Japan rabbits that have been established model of myocardial infarction were randomly divided into 3 groups: the model group, the group with point transplantation of stem cell (0 day) , the group with point transplantation of stem cell (7 days). By the method of thoracotomy with anterior descending coronary artery ligation to establish acute myocardial infarction model, the infarction rabbits were injected with the same volume of separated, cultured, amplified, marked MSCs suspension apartly on 0 day and 7th day. In the five weeks after infarction, immunohistochemical staining Brdu is carried out in myocardial infarction area in rabbits of each group respectively to observe whether the migration of MSCs transplantats to the infarct area. The positive cells in Brdu staining were stained again by cardiac-specific troponin T in order to observe whether the migration of cells differentiate and the nature of differentiation. (2) With the method of TUNEL to determine the number of apoptosis cells, and with the method of SABC, RT-PCR and Western blot to observe the myocardial cells expressing in Fas, FasL, Bcl-2 protein and mRNA.
Results: (1) There were Brdu positive cells and cTnT positive protein in Point injection MSCs group in infarct area. (2) Immunohistochemistry showed: The expression of apoptotic cells in infracted area in model group were more than normal group, the point of stem cell transplantation group(0 day)and the point of stem cell transplantation group (7 days) (P<0. 05), the expression of apoptotic cells in the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05); the expression of Fas protein:the model group was more than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days), the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) ; the expression of FasL protein: the point of stem cell transplantation group(0 day)more than the normal group, the model group and the point of stem cell transplantation group(7 days) (P<0. 05) .The expression of Bcl-2 protein: the model group was less than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days)(P<0. 05) ,the point of stem cell transplantation group(7 days)more than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05). (3) RT-PCR and Western Blotting showed: Compared with the normal group, Fas mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0. 05); FasL mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0.05); Bcl-2 mRNA levels: Model group decreased significantly (P <0.05), the two groups with point injection of stem cells has increased, but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell(7days) also increased , but statistical significance. Compared with model group, Fas mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 01); Bcl-2 mRNA: the two groups with point injection of stem cells increased significantly (P <0. 05) Compared with the group with point transplantation of stem cell(0 day), Fas mRNA levels: stem cells, the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); FASL mRNA levels: the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); Bcl-2 mRNA levels: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0. 05). Compared with the normal group, Fas and FasL protein levels: the two groups with point injection of stem cells was significantly increased (P <0. 05); Bcl-2 protein levels: the model group decreased significantly (P <0. 05), The two groups of acupoint Injection Stem Cells have increased but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell (7 days) was significantly higher(P <0. 05). Compared with model group, Fas protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); Bcl-2 protein: the two groups with point injection of stem cells increased significantly (P <0. 05).Compared with the group with point transplantation of stem cell (0 day), Fas protein: stem cells, the group with point transplantation of stem cell (7days) decreased significantly (P <0. 05) ; FASL protein: the group with point transplantation of stem cell (7 days) decreased significantly (P <0.05) ; Bcl-2 protein: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0.05).
Conclusion: (1) Point transplantation of bone marrow mesenchymal stem cells, can moved to the infarct zone, and can differentiate into myocardial cells; (2) Point transplantation of bone marrow mesenchymal stem cells can reduce the number of cardiomyocyte apoptosis in the infracted and surrounding zone, which may be caused by the reduction of Fas, FasL and the increasing of Bcl-2 to adjustments. (3) Bone marrow-derived mesenchymal stem cells Point-cell transplantation for 7 days can reduce more apoptosis cells than bone marrow-derived mesenchymal stem cells Point-cell transplantation for 0 day.
方法:
1.将已建立心肌梗死模型的日本大耳白兔随机分为3组,模型组、穴位注射干细胞零天组、穴位注射干细胞七天组。用开胸结扎冠状动脉左室支的方法建立急性心肌梗死模型,对梗死后的兔分两组分别在零天与七天穴位注射相同数量的已经分离、培养、扩增、标记后的MSCs悬液。在梗死后五周分别对各组兔心肌梗死区及周边区进行Brdu免疫组化染色以观察穴位注射MSCs是否迁移至梗死区,对Brdu染色阳性的细胞复染心肌特异性肌钙蛋白T染色以观察迁移的细胞是否分化及分化的性质。
2.采用脱氧核苷酸末端转移酶介导原位缺口标记(TUNEL法)确定细胞凋亡数,免疫组化SABC方法及RT-PCR和蛋白印迹法观察心肌细胞中Fas、FasL、Bcl-2蛋白及mRNA的表达。
结果:
1.穴位注射MSCs各组梗死区组织免疫组化Brdu染色阳性;心肌特异性肌钙蛋白T染色阳性。
2.免疫组化示:梗死区凋亡细胞的表达,模型组高于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组(P＜0.05),穴位注射干细胞七天组低于穴位注射干细胞零天组,但无统计学差异(P＞0.05);Fas蛋白的表达,模型组高于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组,穴位注射干细胞七天组低于穴位注射干细胞零天组(P＜0.05);FasL蛋白的表达,穴位注射干细胞零天组高于正常组、模型组、穴位注射干细胞七天组(P＜0.05);Bcl-2蛋白的表达,模型组低于正常组、穴位注射干细胞零天组和穴位注射干细胞七天组(P＜0.05),穴位注射干细胞七天组高于穴位注射干细胞零天组,但无统计学差异(P＞0.05)。
3.RT-PCR与Western Blot显示:与正常组比较,Fas mRNA水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);FasL mRNA水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);Bcl-2 mRNA水平:模型组明显下降(P＜0.05),穴位注射干细胞的两组有所升高,但升高程度不同,穴位注射零天组明显升高(P＜0.05),穴位注射七天组虽有升高但无统计学意义(P＞0.05)。与模型组比较,Fas mRNA水平:穴位注射干细胞的两组明显下降(P＜0.05);FasL mRNA水平:穴位注射干细胞的两组明显下降(P＜0.01);Bcl-2 mRNA:穴位注射干细胞的两组明显升高(P＜0.05)。与穴位注射干细胞零天比较,Fas mRNA水平:穴位注射干细胞七天组明显下降(P＜0.05);FASL mRNA水平:穴位注射干细胞七天组明显下降(P＜0.05);Bcl-2 mRNA水平:穴位注射干细胞七天组也明显下降(P＜0.05)。与正常组比较,Fas与FasL蛋白水平:模型组和穴位注射干细胞的两组明显升高(P＜0.05);Bcl-2蛋白水平:模型组明显下降(P＜0.05),穴位注射干细胞的两组有所升高,但升高程度不同,穴位注射零天组明显升高(P＜0.05),穴位注射七天组明显升高(P＜0.05)。与模型组比较,Fas蛋白:穴位注射干细胞的两组明显下降(P＜0.05);FasL蛋白:穴位注射干细胞的两组明显下降(P＜0.05);Bcl-2蛋白:穴位注射干细胞的两组明显升高(P＜0.05)。与穴位注射干细胞零天比较,Fas蛋白:穴位注射干细胞七天组明显下降(P＜0.05);FASL蛋白:穴位注射干细胞七天组明显下降(P＜0.05);Bcl-2蛋白:穴位注射干细胞七天组也明显下降(P＜0.05)。
结论:
1.穴位注射兔骨髓间充质干细胞可迁移至梗死区,并可以在梗死区分化为类心肌样细胞。
2.穴位注射骨髓间充质干细胞可使梗死及周围区心肌细胞凋亡数减少,其机制可能是通过Fas、FasL蛋白的减少、Bcl-2蛋白的增多来调节的。
3.穴位注射骨髓间充质干细胞七天较穴位注射骨髓间充质干细胞零天更能减少凋亡细胞的数量。
Objective: To investigate the reconstruction capacity of rabbit bone marrow-derived mesenchymal stem cells (MSCs) Point-cell transplantation for ischemic myocyte, at the same time to make a comparative research on cardiomyocyte apoptosis in infarct area between rabbit bone marrow mesenchymal stem cells (MSCs) transplantation Point of zero-day and seven days. To determine the number of cell apoptosis by TUNEL, and to observe the expression of Fas, FasL, Bcl-2 protein and mRNA in myocardial cells by immunohistochemistry SABC methods, RT-PCR and Western blot.
Methods: (1) The Japan rabbits that have been established model of myocardial infarction were randomly divided into 3 groups: the model group, the group with point transplantation of stem cell (0 day) , the group with point transplantation of stem cell (7 days). By the method of thoracotomy with anterior descending coronary artery ligation to establish acute myocardial infarction model, the infarction rabbits were injected with the same volume of separated, cultured, amplified, marked MSCs suspension apartly on 0 day and 7th day. In the five weeks after infarction, immunohistochemical staining Brdu is carried out in myocardial infarction area in rabbits of each group respectively to observe whether the migration of MSCs transplantats to the infarct area. The positive cells in Brdu staining were stained again by cardiac-specific troponin T in order to observe whether the migration of cells differentiate and the nature of differentiation. (2) With the method of TUNEL to determine the number of apoptosis cells, and with the method of SABC, RT-PCR and Western blot to observe the myocardial cells expressing in Fas, FasL, Bcl-2 protein and mRNA.
Results: (1) There were Brdu positive cells and cTnT positive protein in Point injection MSCs group in infarct area. (2) Immunohistochemistry showed: The expression of apoptotic cells in infracted area in model group were more than normal group, the point of stem cell transplantation group(0 day)and the point of stem cell transplantation group (7 days) (P<0. 05), the expression of apoptotic cells in the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05); the expression of Fas protein:the model group was more than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days), the point of stem cell transplantation group(7 days) was less than the point of stem cell transplantation group(0 day) ; the expression of FasL protein: the point of stem cell transplantation group(0 day)more than the normal group, the model group and the point of stem cell transplantation group(7 days) (P<0. 05) .The expression of Bcl-2 protein: the model group was less than the normal group, the point of stem cell transplantation group(0 day) and the point of stem cell transplantation group(7 days)(P<0. 05) ,the point of stem cell transplantation group(7 days)more than the point of stem cell transplantation group(0 day) (but no significant difference P>0. 05). (3) RT-PCR and Western Blotting showed: Compared with the normal group, Fas mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0. 05); FasL mRNA levels: the model group and the two groups with point injection of stem cells was significantly increased (P <0.05); Bcl-2 mRNA levels: Model group decreased significantly (P <0.05), the two groups with point injection of stem cells has increased, but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell(7days) also increased , but statistical significance. Compared with model group, Fas mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL mRNA levels: the two groups with point injection of stem cells decreased significantly (P <0. 01); Bcl-2 mRNA: the two groups with point injection of stem cells increased significantly (P <0. 05) Compared with the group with point transplantation of stem cell(0 day), Fas mRNA levels: stem cells, the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); FASL mRNA levels: the group with point transplantation of stem cell (7 days) decreased significantly (P <0. 05); Bcl-2 mRNA levels: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0. 05). Compared with the normal group, Fas and FasL protein levels: the two groups with point injection of stem cells was significantly increased (P <0. 05); Bcl-2 protein levels: the model group decreased significantly (P <0. 05), The two groups of acupoint Injection Stem Cells have increased but increased in varying degrees, the group with point transplantation of stem cell (0 day) was significantly higher (P <0. 05), the group with point transplantation of stem cell (7 days) was significantly higher(P <0. 05). Compared with model group, Fas protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); FasL protein: the two groups with point injection of stem cells decreased significantly (P <0. 05); Bcl-2 protein: the two groups with point injection of stem cells increased significantly (P <0. 05).Compared with the group with point transplantation of stem cell (0 day), Fas protein: stem cells, the group with point transplantation of stem cell (7days) decreased significantly (P <0. 05) ; FASL protein: the group with point transplantation of stem cell (7 days) decreased significantly (P <0.05) ; Bcl-2 protein: the group with point transplantation of stem cell (7 days) was also significantly decreased (P <0.05).
Conclusion: (1) Point transplantation of bone marrow mesenchymal stem cells, can moved to the infarct zone, and can differentiate into myocardial cells; (2) Point transplantation of bone marrow mesenchymal stem cells can reduce the number of cardiomyocyte apoptosis in the infracted and surrounding zone, which may be caused by the reduction of Fas, FasL and the increasing of Bcl-2 to adjustments. (3) Bone marrow-derived mesenchymal stem cells Point-cell transplantation for 7 days can reduce more apoptosis cells than bone marrow-derived mesenchymal stem cells Point-cell transplantation for 0 day.
引文
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[15]Jackson KA,Majka SM,Wang H,etal.Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells[J].J Clin Invest,2001,107(11):1355-1356.
[16]Chen SL,Fang WW,Qian J,etal.Improvement of cardiac function after transplantation of autologous bone marrow mesenchymal stem cells in patients with acute myocardial infarction[J].Chin Med J,2004,117(10):1443-1448.
[17]Strauer BE,Kornowski R.Stem cell therapy in perspective[J].Circulation,2003,107(7):929-934.
[18]Orlic D,Kajstura J,Chimenti S,etal.Bone marrow cells regenerate infarcted myocardium[J].Pediatr Transplant,2003,3:86-88.
[19]Timmermans F,De Sutter J,Gillebert TC.Stem cells for the heart,are we there yet?[J]Cardiology,2003,100(4):176-185.
[20]Assmus B.Schachinger V.Teupe C.etal.Transplantation of progenitor cells and regeneration enhancement in acute myocardialin faction[J].Circulation,2002,106(24),3009-3017.
[21]Janssens S,Dubois C,Bogaert J,etal.Autologous bone marrow-derived stem cell transfer in patients with ST-segment elevation myocardial infarction: double-blind randomised controlled trial[J].Lancet,2006,367(9505),1:113-121.
[22]Lunde K,Solheim S,Aakhus S,etal.Autogous Stem cell transplantation in acute myocardial infarction:The ASTAM1 randomized controlled trial[7].Scand Cardiovasc J,2005,39(3),150-158.
[23]Kuethe F,Richartz BM,Sayer HG,etal.Lack of regeneration of myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans with large anterior myocardial infarctions[J].Cardiol,2004,97(1):123-127.
[24]Schachinger V,Assmus B,Britten MB.etal.Transplantation of progenitor cells and regeneration enhancement in acute myocardial infarction:final oneyear results of the TOPCARE AM1 Tria[J].J Am Coll Cardiol,2004,44(8),1690-1699.
[25]杨水祥,胡大一.自体骨髓干细胞移植治疗心肌梗死6例报告[J].中国临床康复,2005,9(46):11-13.
[26]任福强,朱玲,石应康.骨骼肌卫星细胞对心肌梗死的治疗[J].四川生理科学,2007,02,9(2):11.
[27]Xu W,Zhang X,Qian H,etal.Mesenchymal stem cells from adult human bone marrow differentiate into a cardiomyocyte phenotype in vitro[J].Exp Biol Med,2004,229(7):623-631.
[28]Shim WS,Jiang S,etal.Ex vivo differentiation of human adult bone marrow stem cells into cardiomyocyte-like cells[J].Biochem Biophys Res Commun,2004,324(2):481-488.
[29]Tendera M,Wojakowski W.Clinical trials using autologous bone marrow and peripheral blood-derived progenitor cells in patients with acute myocardial infarction[J].Folia Histochem Cytobiol,2005,43(4):233-235.
[30]Chang SA,Kim HK,hee HY,etal.Restoration of the left ventricular synchronous contraction after acute myocardial infarction by stem cell therapy:new insights into the therapeutic implication of stem cell therapy for acute myocardial infarction[J].Heart,2008,94(8):969-970.
[31]Feygin J,Mansoor A,Eckman P,etal.Functional and bioenergetic modulations in the infarct border zone following autologous mesenchymal stem cell transplantation [J].Am J Physiol Heart Circ Physiol, 2007, 293(3):H1772-80.
[32]Grauss RW, Winter EM, Van Tuvn J, etal. Mesenchymal stem cells from ischemic heart disease patients improve left ventricular function after acute myocardial infarction[J]. Am J Physiol Heart Circ Physiol, 2007, 293(4):H2438-47.
[33]Shyu KG, Wang BW, J Biomed. Mesenchymal stem cells are superior to angiogenic growth factor genes for improving myocardial performance in the mouse model of acute myocardial infarction[J]. Biomed Sci, 2006 Jan, 13(1):47-58.
[34]Schuleri KH, Boyle AJ, Hare JM. Mesenchymal stem cells for cardiac regenerative therapy[J]. Handb Exp Pharmacol, 2007, (180):195-218.
[35]Abdel-Latif A, Bolli R, Tleyjeh IM, etal. Adult bone marrow-derived cells for cardiac repair:a systematic review and meta-analysis[J], Arch Intern Med, 2007,28, 167(10): 989-97.
[36]Vassalli G, Vanderheyden M, Renders F, etal. Bone marrow stem cell therapy for cardiac repair: challenges and perspectives[J]. Minerva Cardioangiol, 2007, 55(5): 659-667.
[37]Tatsumi T, Matsubara H. Therapeutic angiogenesis for peripheral arterial disease and ischemic heart disease by autologous bone marrow cells implantation [J]. Nippon Rinsho, 2006, 64(11):2126-2134.
[38]Menasche P. Perspectives in cardiac cell therapy[J]. Presse Med, 2007,361: 1S55-8.
[39]Gulati R, Simari RD. Cell therapy for acute myocardial infarction[J]. Med Clin North Am, 2007, 91 (4):769-785.
[40]Guo J, Lin G, Bao C, etal. Insulin-like growth factor 1 improves the efficacy of mesenchymal stem cells transplantation in a rat model of myocardial infarction. Biomed Sci, 2008, 15(1):89-97.
[41]YangJ, Zhou W, Zheng W, etal. Effects of myocardial transplantation of marrow mesenchymal stem cells transfected with vascular endothelial growth factor for the improvement of heart function and angiogenesis after myocardial infarction [J]. Cardiology, 2007, 107(1):17-29.
[42]Chen J,Baydoun AR,Xu R,etal.Lysophosphatidic Acid Protects Mesenchymal Stem Cells against Hypoxia and Serum Deprivation-Induced Apoptosis[J].Stem Cells,2008,26(1):135-145.
[43]Canepa M,Coviello D,Chiarella F.Cardiac cell therapy:the puzzle is waiting to be solved[J].G Ital Cardiol(Rome),2006,7(4):252-265.
[45]Tendera M,Wojakowski W.Clinical trials using autologous bone marrow and peripheral blood-derived progenitor cells in patients with acute myocardial infarction[J].Folia Histochem Cytobiol,2005,43(4):233-235.
[1]倪峰,林静瑜等.穴位注射疗法作用机制探讨[J].中国针灸,2003,23(10):609-611.
[2]杨丹红.穴位注射对高脂血症动物模型的降脂作用研究[J].针灸临床杂志,2000,16(1):43-45.
[3]徐桂芬,王文华,姚凤祯.穴位注射生脉注射液抗家兔快速性心律失常的实验研究[J]。中国急救医学,2006,10(26):683-684.
[4]张慧.药物穴位注射对冠心病ST-T段的即时影响[J].临床医学,2007,27(8):17.
[5]宋建华,李斌.穴位注射配合中药治疗冠心病心绞痛58例[J].上海针灸杂志,2006,25(10):16-17.
[6]王全权,陈海林,黄慧敏.复方丹参注射液治疗冠心病心绞痛的疗效观察[J].时珍国医国药,2006,17(1):90-91.
[7]张建平,王焕玲.穴位注射治疗冠心病心绞痛的临床观察[J].湖北中医杂志,2002,24(12):42.
[8]毛喜荣,李佩芳,王月兰.穴位注射治疗冠状动脉粥样硬化性心脏病102例的临床观察[J].针灸临床杂志,1995,11(2):16-18.
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[16]Chen SL,Fang WW,Qian J,etal.Improvement of cardiac function after transplantation of autologous bone marrow mesenchymal stem cells in patients with acute myocardial infarction[J].Chin Med J,2004,117(10):1443-1448.
[17]Strauer BE,Kornowski R.Stem cell therapy in perspective[J].Circulation,2003,107(7):929-934.
[18]Orlic D,Kajstura J,Chimenti S,etal.Bone marrow cells regenerate infarcted myocardium[J].Pediatr Transplant,2003,3:86-88.
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[23]Kuethe F,Richartz BM,Sayer HG,etal.Lack of regeneration of myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in humans with large anterior myocardial infarctions[J].Cardiol,2004,97(1):123-127.
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[35]Abdel-Latif A, Bolli R, Tleyjeh IM, etal. Adult bone marrow-derived cells for cardiac repair:a systematic review and meta-analysis[J], Arch Intern Med, 2007,28, 167(10): 989-97.
[36]Vassalli G, Vanderheyden M, Renders F, etal. Bone marrow stem cell therapy for cardiac repair: challenges and perspectives[J]. Minerva Cardioangiol, 2007, 55(5): 659-667.
[37]Tatsumi T, Matsubara H. Therapeutic angiogenesis for peripheral arterial disease and ischemic heart disease by autologous bone marrow cells implantation [J]. Nippon Rinsho, 2006, 64(11):2126-2134.
[38]Menasche P. Perspectives in cardiac cell therapy[J]. Presse Med, 2007,361: 1S55-8.
[39]Gulati R, Simari RD. Cell therapy for acute myocardial infarction[J]. Med Clin North Am, 2007, 91 (4):769-785.
[40]Guo J, Lin G, Bao C, etal. Insulin-like growth factor 1 improves the efficacy of mesenchymal stem cells transplantation in a rat model of myocardial infarction. Biomed Sci, 2008, 15(1):89-97.
[41]YangJ, Zhou W, Zheng W, etal. Effects of myocardial transplantation of marrow mesenchymal stem cells transfected with vascular endothelial growth factor for the improvement of heart function and angiogenesis after myocardial infarction [J]. Cardiology, 2007, 107(1):17-29.
[42]Chen J,Baydoun AR,Xu R,etal.Lysophosphatidic Acid Protects Mesenchymal Stem Cells against Hypoxia and Serum Deprivation-Induced Apoptosis[J].Stem Cells,2008,26(1):135-145.
[43]Canepa M,Coviello D,Chiarella F.Cardiac cell therapy:the puzzle is waiting to be solved[J].G Ital Cardiol(Rome),2006,7(4):252-265.
[45]Tendera M,Wojakowski W.Clinical trials using autologous bone marrow and peripheral blood-derived progenitor cells in patients with acute myocardial infarction[J].Folia Histochem Cytobiol,2005,43(4):233-235.
[1]倪峰,林静瑜等.穴位注射疗法作用机制探讨[J].中国针灸,2003,23(10):609-611.
[2]杨丹红.穴位注射对高脂血症动物模型的降脂作用研究[J].针灸临床杂志,2000,16(1):43-45.
[3]徐桂芬,王文华,姚凤祯.穴位注射生脉注射液抗家兔快速性心律失常的实验研究[J]。中国急救医学,2006,10(26):683-684.
[4]张慧.药物穴位注射对冠心病ST-T段的即时影响[J].临床医学,2007,27(8):17.
[5]宋建华,李斌.穴位注射配合中药治疗冠心病心绞痛58例[J].上海针灸杂志,2006,25(10):16-17.
[6]王全权,陈海林,黄慧敏.复方丹参注射液治疗冠心病心绞痛的疗效观察[J].时珍国医国药,2006,17(1):90-91.
[7]张建平,王焕玲.穴位注射治疗冠心病心绞痛的临床观察[J].湖北中医杂志,2002,24(12):42.
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