心血瘀阻证心肌微环境变化及对骨髓间充质干细胞移植的影响
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摘要
目的：
研究心血瘀阻证心肌微环境对骨髓间充质干细胞(bone mesenchymal stem cells BMSCs)向心肌样分化的作用及养心通脉有效部位方(active principle region of Yangxin Tongmai Formula apr-YTF)对其影响。
方法：
(1)理论探讨：通过对“微环境(microenvironment)"概念,发展历史,及当前学术趋势的检索,从理论上探讨微环境的概念、学术发展趋势、心血瘀阻证心肌微环境的变化特点及其对BMSCs移植的影响,并为从改善微环境的角度寻求中医药干预BMSCs向心肌分化提供理论依据。
(2)实验研究：①建立大鼠急性心梗心血瘀阻证模型,随机分为A、B两部分,A部分取大鼠心肌组织,用ELISA方法观测其基质金属蛋白酶2(matrix metalloproteinase-2 MMP-2)、基质金属蛋白酶9 (matrix metalloproteinase-9 MMP-9)的表达；用Western-Blots方法检测细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。B部分动物模型在梗死心肌部位移植BMSCs观测其上述指标；同时用免疫组化双染的方法观测梗死心肌组织中结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析心肌分化指标和MMP-2、MMP-9,信号转导通路PI3K、p38相关蛋白表达的相关性。②建立大鼠急性心梗心血瘀阻证模型,在梗死心肌部位移植BMSCs;并用apr-YTF进行干预。用ELISA方法观测心肌组织中MMP-2、MMP-9的表达；用Western-Blots方法检测心肌组织中细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。同时用免疫组化双染的方法观测其心肌结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析上述心肌分化指标和MMP-2、MMP-9、信号转导通路PI3K、p38中相关蛋白的相关性。③体外模拟急性心梗心血瘀阻证微环境,在此微环境下培养BMSCs并用apr-YTF进行干预。用ELISA方法观测其培养液中MMP-2、MMP-9的表达；用Western-Blots方法检测细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。同时用免疫细胞化学方法观测其心肌结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析上述心肌分化指标和MMP-2、MMP-9,信号转导通路PI3K、p38中相关蛋白的相关性。
结果：
(1)接受BMSCs移植后可见心血瘀阻证心肌梗死边缘区有Desmin-Brdu、MHC-Brdu两种蛋白的同时表达,而在假手术组和健康对照组未见双染细胞表达。GATA-4 mRNA心血瘀阻证组高于假手术组(P＜0.05),而健康对照组的心肌中未检出表达。
(2)未作BMSCs移植各部分MMP-2心血瘀阻证表达高于健康对照和假手术组(P＜0.05),接受BMSCs移植后大鼠心肌组织各组间未见统计学差异(P＞0.05)；相同组别内部比较心血瘀阻证组移植BMSCs部分表达低于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各部分MMP-9三组间均有差异,其中心血瘀阻证组最高,其次为假手术组、健康对照组(P＜0.05),接受BMSCs移植后心血瘀阻证组和假手术组低于健康对照组(P＜0.05),相同组别内部比较心血瘀阻证组移植BMSCs部分表达低于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各组大鼠心肌组织p-Akt统计学未见明显差异(P＞0.05),接受BMSCs移植心血瘀阻证表达高于健康对照和假手术组(P＜0.05),相同组别内部比较心血瘀阻证组和健康对照组中移植BMSCs部分表达高于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各组p-p38数据统计学未见明显差异(P>0.05),接受BMSCs移植后各组大鼠心肌组织,各组之间未见差别(P＞0.05),相同组别内部比较心血瘀阻证组中移植BMSCs部分表达高于未作BMSCs移植部分(P＜0.05)。各组标本GATA-4 mRNA与MMP-9、p-p38含量表达呈正相关,而与MMP-2、p-Akt未见显相关性。
(3)apr-YTF干预BMSCs移植心血瘀阻证心肌后经心肌结蛋白Desmin-Brdu抗体染色后,其中(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组阳性细胞计数均高于NS+BMSCs组(P＜0.05),前两者之间无差别(P＞0.05)。MHC-Brdu抗体染色后,(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组阳性细胞计数与高于NS+BMSCs组(P＜0.05),前两者之间无差别(P＞0.05)。GATA-4 mRNA IOD值半定量分析NS+BMSCs组、(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组均高于NS+DMEM组,(apr-YTF)+BMSCs组高于NS+BMSCs组(P＜0.05)。
(4) apr-YTF干预BMSCs移植心血瘀阻证心肌酶联免疫吸附法检测显示MMP-2 (apr-YTF)+BMSCs组表达低于NS+DMEM组和NS+BMSCs组(P＜0.05和P＜0.01)。MMP-9各组间未见差异(P＞0.05)。经对Western-Blots法检测p-Akt (apr-YTF)+BMSCs组表达高于NS+DMEM组、NS+BMSCs组和(rhG-CSF)+BMSCs组(P＜0.05)。p-p38 (apr-YTF)+BMSCs组表达高于NS+DMEM (P＜0.05),其余各组间未见统计学差异(P＜0.05)。
(5)体外模拟心血瘀阻证微环境对BMSCs诱导后,在正常对照组和DMEM组中未发现Desmin及MHC阳性细胞；心血瘀阻证组、apr-YTF组和心血瘀阻证+(apr-YTF)组Desmin及MHC免疫组化染色均可见弱阳性免疫复合物。Desmin的表达的IOD值在在心血瘀阻证组和心血瘀阻证+(apr-YTF)组均高于apr-YTF组(P＜0.05)。MHC的表达的OD值在在心血瘀阻证组和心血瘀阻证+(apr-YTF)组均高于apr-YTF组(P＜0.05),但是前两者之间无统计学差异。心血瘀阻证组和心血瘀阻证+(apr-YTF)组GATA-4 mRNA相对光密度值与正常对照组比较差异具有统计学意义(P＜0.05和P＜0.01)。
(6)体外模拟心血瘀阻证微环境细胞培养上清液MMP-2的质量浓度以心血瘀阻证+(apr-YTF)组含量最高,与正常对照组、DMEM组和apr-YTF组比较有统计学差异(P＜0.05)。MMP-9的质量浓度心血瘀阻证组和心血瘀阻证+(apr-YTF)组两个组高于DMEM组和apr-YTF组(P＜0.05)。BMSCs诱导28d后,信号转导通路中p-Akt以心血瘀阻证+(apr-YTF)组含量最高,与DMEM组、心血瘀阻证组和apr-YTF组比较周统计学差异(P＜0.05)。而p-p38各组间含量表达未见统计学差异(P＜0.05)
结论：
(1)心梗心血瘀阻证心肌微环境可以促进BMSCs向心肌样细胞分化；apr-YTF可以起到促进作用。
(2)心梗心血瘀阻证心肌微环境中MMP-2、MMP-9表达增高,促使BMSCs迁移分化。BMSCs移植后心血瘀阻证心肌微环境中MMP-2、MMP-9含量表达降低,从而减轻组织降解,瘢痕形成,控制心梗后向心衰的发展。apr-YTF可以降低心梗心血瘀阻证心肌组织中MMP-2、MMP-9表达。
(3)心梗心血瘀阻证心肌微环境中PI3K信号转导通路激活。BMSCs移植后心肌微环境中心肌样细胞分化及与PI3K及p38信号转导通路激活有关。apr-YTF能促使移植的BMSCs向心肌样细胞分化,其分化机制与PI3K及p38信号转导通路激活有关。
(4)体外模拟的心血瘀阻证微环境可以诱导BMSCs向心肌样细胞分化。apr-YTF在体外可以诱导BMSCs向心肌样细胞分化。
(5)体外模拟的心血瘀阻证微环境MMP-2、MMP-9可以促使BMSCs向心肌样细胞分化。在体外模拟的心血瘀阻证微环境中BMSCs向心肌样细胞分化及apr-YTF的干预作用与PI3K信号转导通路激活有关。
Objective:
To study the function about cardiac blood stasis syndrome cardiac microenvironment towards BMSCs cardiac differentiation as well as the influences about the active principle region of Yangxin Tongmai Formula(apr-YTF).
Methods:
(1)Theoretical discussion:Through revieve the concept, development history and current academic tendency of microenvironment, to explore its exact concept academic development tendency and change characteristics of cardiac blood stasis syndrome and its influences on the bone marrow mesenchyma stem cell(BMSCs) transplant. To provide theoretical basis for TCM interferences on BMSCs transplant into cardiac differentiation.
(2)Experimental research:①Established the model of rats acute myocardial infarction (AMI) cardiac blood stasis syndrome, use ELISA method to observe the expression about matalloprotease-2 (MMP-2) and matalloprotease-9 (MMP-9);use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K and p38, meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC;Use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the index of cellular differentiation as well as the MMP-2 and MMP-9 and associativity of signal transduction PI3K、p38.②Established the model of rats AMI cardiac blood stasis syndrome and transplanted BMSCs in infarctus cardiac muscule, use ELISA method to observe the expression about MMP-2 and MMP-9, use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K and p38;meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC, use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the associativity between the index of cellular differentiation as well as the MMP-2 and MMP-9 and content of signal transduction PI3K、p38.③Simulated the environment about AMI cardiac blood stasis syndrome in vitro, to cultivate the BMSCs and interfere by apr-YTF, use ELISA method to observe the expression about MMP-2 and MMP-9, use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K、p38, meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC;Use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the ssociativity between the index of cellular differentiation as well as the MMP-2 and MMP-9 and content of signal transduction PI3K、p38.
Results:
(1)After BMSCs transplant there was simultaneous expression of Desmin-Brdu、MHC-Brdu in the cardiac blood stasis infarctus edge area. There was no expression in the sham operation group and health control group. GATA-4 mRNA in the cardiac blood stasis group was higher than that in the sham operation group(P＜0.05),while there was no expression in the health control group.
(2)Every part of no BMSCs transplant of MMP-2 cardiac blood stasis syndrome was higher that of sham operation group and health control group. After BMSCs transplant there was statistical difference of rats cardiac tissues between each groups (P＜0.05). The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). Every part of no BMSCs transplant of MMP-9 had statistical differences between each group. Cardiac blood stasis syndrome group was the highest, sham operation group was the next one and health control group was the last one. After BMSCs transplant, the cardiac blood stasis syndrome group was lower than that of health control group and sham operation group. The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). Every part of no BMSCs transplant of p-Akt had no statistical differences between each group.After BMSCs transplant, the cardiac blood stasis syndrome group was lower than that of health control group and sham operation group, the expression of transplant group's was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05).
Every part of no BMSCs transplant of p-p38 had no statistical differences between each group. There was no difference between each group's rats cardiac tissues after BMSCs transplant. The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). The expression of GATA-4 mRNA,MMP-9 and p-p38 was positive correlation. The incidence coefficient were 0.49and 0.605 while there was no obvious associativity of MMP-2、p-Akt.
(3)Apr-YTF interfered in BMSCs transplant of cardiac blood stasis of Desmin-Brdu, the (rhG-CSF)+BMSCs group's positive cell counting were higher than that of NS+BMSCs group. There was no difference between (rhG-CSF)+BMSCs group and NS+BMSCs group. Through MHC-Brdu antibody dyeing, (rhG-CSF)+BMSCs and (apr-YTF)+BMSCs group's positive cell counting were higher than that of NS+BMSCs group. The semi-quantitative IOD of GATA-4 mRNA, NS+BMSCs group as well as (rhG-CSF)+BMSCs and (apr-YTF)+BMSCs group were higher than that of NS+DMEM group;while the (apr-YTF)+BMSCs group was higher than that of NS+BMSCs.
(4)Apr-YTF interfered in BMSCs transplant of cardiac blood stasis of MMP-2 and MMP-9, ELISA test showed that the MMP-2 (apr-YTF)+BMSCs group's expression was lower than that of NS+BMSCs group and NS+BMSCs. There was no difference between each group of MMP-9. Through p-Akt Western-Blot, (apr-YTF)+BMSCs's expression was higher than that of in NS+DMEM group NS+BMSCs group as well as (rhG-CSF)+BMSCs group. The p-p38 (apr-YTF)+BMSCs group's expression was higher than that of NS+DMEM group. There was no difference between each other group.
(5)After the induction of BMSCs simulated the environment about AMI cardiac blood stasis syndrome in vitro, there was no MHC positive cell been detected in the normal control group and DMEM group, while the weakly positive immunocomplex had been detected in the cardiac blood stasis syndrome group, apr-YTF group, cardiac blood stasis syndrome+apr-YTF in the Desmin and MHC immunohistochemistry dyeing. The IOD of Desmin expressionin the cardiac blood stasis syndrome group and cardiac blood stasis syndrome+(apr-YTF)group were higher than in the apr-YTF group, but there was no statistical difference between the former two ones。The GATA-4 mRNA IOD in cardiac blood stasis syndrome group and cardiac blood stasis syndrome+(apr-YTF)group had statistical differences compared with the normal control group.
(6)The MMP-2's mass concentration in the cell culture of simulated the environment of cardiac blood stasis syndrome+apr-YTF group was the highest which had the statistical difference compared with the normal control group DMEM and apr-YTF group. The mass concentration of MMP-9 in the two cardiac blood stasis syndrome group, and cardiac blood stasis syndrome+apr-YTF group were higher than in the DMEM and apr-YTF group.After 28 days of BMSCs' induction, the p-Akt in the signal transduction path was much higher in the cardiac blood stasis syndrome+apr-YTF group than that in the DMEM, apr-YTF and cardiac blood stasis syndrome group. The p-p38's expression in each group had no statistical difference.
Conclusion:
(1)The microenvironment of myocardial infarction cardiac blood stasis syndrome's cardiac cell can promote the BMSCs cardiac cell cytodifferentiation. Apr-YTF can promote it.
(2)The microenvironment of myocardial infarction cardiac blood stasis syndrome's cardiac cell can increase the expression of MMP-2 and MMP-9, promote the BMSCs migration and differentiation. After BMSCs transplant cardiac blood stasis syndrome's cardiac cell microenvironment's content expression lower and thus mitigate the tissure degradation cicatrization and control the deterioration of MI. Apr-YTF can lower the MMP's expression of AMI cardiac blood stasis syndrome in cardian tissure.
(3)PI3K signal transduction path in cardiac blood stasis syndrome's cardiac cell microenvironment has been activated. After BMSCs transplant cardiac blood stasis syndrome's cardiac cell microenvironment's cardiac cell differentiation related to the activation of PI3K and p38 signal transduction. Apr-YTF can promote the BMSCs transplant into cardiac cell differentiation and the activation of PI3K and p38 signal transduction.
(4)Simulated the microenvironment about cardiac blood stasis syndrome in vitro can induce the BMSCs cardiac cell differentiation. Apr-YTF can induce the BMSCs cardiac cell differentiation in vitro.
(5)Simulated the microenvironment about cardiac blood stasis syndrome in vitro, MMP-2 and MMP-9 can promote the BMSCs cardiac cell differentiation. BMSCs differentiation to cardiac cell and apr-YTF's interferences has related to the activation of PI3K signal transduction path.
研究心血瘀阻证心肌微环境对骨髓间充质干细胞(bone mesenchymal stem cells BMSCs)向心肌样分化的作用及养心通脉有效部位方(active principle region of Yangxin Tongmai Formula apr-YTF)对其影响。
方法：
(1)理论探讨：通过对“微环境(microenvironment)"概念,发展历史,及当前学术趋势的检索,从理论上探讨微环境的概念、学术发展趋势、心血瘀阻证心肌微环境的变化特点及其对BMSCs移植的影响,并为从改善微环境的角度寻求中医药干预BMSCs向心肌分化提供理论依据。
(2)实验研究：①建立大鼠急性心梗心血瘀阻证模型,随机分为A、B两部分,A部分取大鼠心肌组织,用ELISA方法观测其基质金属蛋白酶2(matrix metalloproteinase-2 MMP-2)、基质金属蛋白酶9 (matrix metalloproteinase-9 MMP-9)的表达；用Western-Blots方法检测细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。B部分动物模型在梗死心肌部位移植BMSCs观测其上述指标；同时用免疫组化双染的方法观测梗死心肌组织中结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析心肌分化指标和MMP-2、MMP-9,信号转导通路PI3K、p38相关蛋白表达的相关性。②建立大鼠急性心梗心血瘀阻证模型,在梗死心肌部位移植BMSCs;并用apr-YTF进行干预。用ELISA方法观测心肌组织中MMP-2、MMP-9的表达；用Western-Blots方法检测心肌组织中细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。同时用免疫组化双染的方法观测其心肌结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析上述心肌分化指标和MMP-2、MMP-9、信号转导通路PI3K、p38中相关蛋白的相关性。③体外模拟急性心梗心血瘀阻证微环境,在此微环境下培养BMSCs并用apr-YTF进行干预。用ELISA方法观测其培养液中MMP-2、MMP-9的表达；用Western-Blots方法检测细胞分化相关信号转导通路PI3K、p38中相关蛋白的变化。同时用免疫细胞化学方法观测其心肌结蛋白Desmin、肌球蛋白重链MHC的表达,用RT-PCR的方法检测心肌转录因子GATA-4 mRNA的表达,然后分析上述心肌分化指标和MMP-2、MMP-9,信号转导通路PI3K、p38中相关蛋白的相关性。
结果：
(1)接受BMSCs移植后可见心血瘀阻证心肌梗死边缘区有Desmin-Brdu、MHC-Brdu两种蛋白的同时表达,而在假手术组和健康对照组未见双染细胞表达。GATA-4 mRNA心血瘀阻证组高于假手术组(P＜0.05),而健康对照组的心肌中未检出表达。
(2)未作BMSCs移植各部分MMP-2心血瘀阻证表达高于健康对照和假手术组(P＜0.05),接受BMSCs移植后大鼠心肌组织各组间未见统计学差异(P＞0.05)；相同组别内部比较心血瘀阻证组移植BMSCs部分表达低于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各部分MMP-9三组间均有差异,其中心血瘀阻证组最高,其次为假手术组、健康对照组(P＜0.05),接受BMSCs移植后心血瘀阻证组和假手术组低于健康对照组(P＜0.05),相同组别内部比较心血瘀阻证组移植BMSCs部分表达低于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各组大鼠心肌组织p-Akt统计学未见明显差异(P＞0.05),接受BMSCs移植心血瘀阻证表达高于健康对照和假手术组(P＜0.05),相同组别内部比较心血瘀阻证组和健康对照组中移植BMSCs部分表达高于未作BMSCs移植部分(P＜0.05)。未作BMSCs移植各组p-p38数据统计学未见明显差异(P>0.05),接受BMSCs移植后各组大鼠心肌组织,各组之间未见差别(P＞0.05),相同组别内部比较心血瘀阻证组中移植BMSCs部分表达高于未作BMSCs移植部分(P＜0.05)。各组标本GATA-4 mRNA与MMP-9、p-p38含量表达呈正相关,而与MMP-2、p-Akt未见显相关性。
(3)apr-YTF干预BMSCs移植心血瘀阻证心肌后经心肌结蛋白Desmin-Brdu抗体染色后,其中(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组阳性细胞计数均高于NS+BMSCs组(P＜0.05),前两者之间无差别(P＞0.05)。MHC-Brdu抗体染色后,(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组阳性细胞计数与高于NS+BMSCs组(P＜0.05),前两者之间无差别(P＞0.05)。GATA-4 mRNA IOD值半定量分析NS+BMSCs组、(rhG-CSF)+BMSCs组和(apr-YTF)+BMSCs组均高于NS+DMEM组,(apr-YTF)+BMSCs组高于NS+BMSCs组(P＜0.05)。
(4) apr-YTF干预BMSCs移植心血瘀阻证心肌酶联免疫吸附法检测显示MMP-2 (apr-YTF)+BMSCs组表达低于NS+DMEM组和NS+BMSCs组(P＜0.05和P＜0.01)。MMP-9各组间未见差异(P＞0.05)。经对Western-Blots法检测p-Akt (apr-YTF)+BMSCs组表达高于NS+DMEM组、NS+BMSCs组和(rhG-CSF)+BMSCs组(P＜0.05)。p-p38 (apr-YTF)+BMSCs组表达高于NS+DMEM (P＜0.05),其余各组间未见统计学差异(P＜0.05)。
(5)体外模拟心血瘀阻证微环境对BMSCs诱导后,在正常对照组和DMEM组中未发现Desmin及MHC阳性细胞；心血瘀阻证组、apr-YTF组和心血瘀阻证+(apr-YTF)组Desmin及MHC免疫组化染色均可见弱阳性免疫复合物。Desmin的表达的IOD值在在心血瘀阻证组和心血瘀阻证+(apr-YTF)组均高于apr-YTF组(P＜0.05)。MHC的表达的OD值在在心血瘀阻证组和心血瘀阻证+(apr-YTF)组均高于apr-YTF组(P＜0.05),但是前两者之间无统计学差异。心血瘀阻证组和心血瘀阻证+(apr-YTF)组GATA-4 mRNA相对光密度值与正常对照组比较差异具有统计学意义(P＜0.05和P＜0.01)。
(6)体外模拟心血瘀阻证微环境细胞培养上清液MMP-2的质量浓度以心血瘀阻证+(apr-YTF)组含量最高,与正常对照组、DMEM组和apr-YTF组比较有统计学差异(P＜0.05)。MMP-9的质量浓度心血瘀阻证组和心血瘀阻证+(apr-YTF)组两个组高于DMEM组和apr-YTF组(P＜0.05)。BMSCs诱导28d后,信号转导通路中p-Akt以心血瘀阻证+(apr-YTF)组含量最高,与DMEM组、心血瘀阻证组和apr-YTF组比较周统计学差异(P＜0.05)。而p-p38各组间含量表达未见统计学差异(P＜0.05)
结论：
(1)心梗心血瘀阻证心肌微环境可以促进BMSCs向心肌样细胞分化；apr-YTF可以起到促进作用。
(2)心梗心血瘀阻证心肌微环境中MMP-2、MMP-9表达增高,促使BMSCs迁移分化。BMSCs移植后心血瘀阻证心肌微环境中MMP-2、MMP-9含量表达降低,从而减轻组织降解,瘢痕形成,控制心梗后向心衰的发展。apr-YTF可以降低心梗心血瘀阻证心肌组织中MMP-2、MMP-9表达。
(3)心梗心血瘀阻证心肌微环境中PI3K信号转导通路激活。BMSCs移植后心肌微环境中心肌样细胞分化及与PI3K及p38信号转导通路激活有关。apr-YTF能促使移植的BMSCs向心肌样细胞分化,其分化机制与PI3K及p38信号转导通路激活有关。
(4)体外模拟的心血瘀阻证微环境可以诱导BMSCs向心肌样细胞分化。apr-YTF在体外可以诱导BMSCs向心肌样细胞分化。
(5)体外模拟的心血瘀阻证微环境MMP-2、MMP-9可以促使BMSCs向心肌样细胞分化。在体外模拟的心血瘀阻证微环境中BMSCs向心肌样细胞分化及apr-YTF的干预作用与PI3K信号转导通路激活有关。
Objective:
To study the function about cardiac blood stasis syndrome cardiac microenvironment towards BMSCs cardiac differentiation as well as the influences about the active principle region of Yangxin Tongmai Formula(apr-YTF).
Methods:
(1)Theoretical discussion:Through revieve the concept, development history and current academic tendency of microenvironment, to explore its exact concept academic development tendency and change characteristics of cardiac blood stasis syndrome and its influences on the bone marrow mesenchyma stem cell(BMSCs) transplant. To provide theoretical basis for TCM interferences on BMSCs transplant into cardiac differentiation.
(2)Experimental research:①Established the model of rats acute myocardial infarction (AMI) cardiac blood stasis syndrome, use ELISA method to observe the expression about matalloprotease-2 (MMP-2) and matalloprotease-9 (MMP-9);use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K and p38, meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC;Use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the index of cellular differentiation as well as the MMP-2 and MMP-9 and associativity of signal transduction PI3K、p38.②Established the model of rats AMI cardiac blood stasis syndrome and transplanted BMSCs in infarctus cardiac muscule, use ELISA method to observe the expression about MMP-2 and MMP-9, use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K and p38;meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC, use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the associativity between the index of cellular differentiation as well as the MMP-2 and MMP-9 and content of signal transduction PI3K、p38.③Simulated the environment about AMI cardiac blood stasis syndrome in vitro, to cultivate the BMSCs and interfere by apr-YTF, use ELISA method to observe the expression about MMP-2 and MMP-9, use Western-Blots method examine the change about the protein of cellular differentiation signal transduction path PI3K、p38, meanwhile use the immunohistochemistry double-dyed method to measure the Desmin and MHC;Use RT-PCR method to observe the expression about GATA-4 mRNA, and analyze the ssociativity between the index of cellular differentiation as well as the MMP-2 and MMP-9 and content of signal transduction PI3K、p38.
Results:
(1)After BMSCs transplant there was simultaneous expression of Desmin-Brdu、MHC-Brdu in the cardiac blood stasis infarctus edge area. There was no expression in the sham operation group and health control group. GATA-4 mRNA in the cardiac blood stasis group was higher than that in the sham operation group(P＜0.05),while there was no expression in the health control group.
(2)Every part of no BMSCs transplant of MMP-2 cardiac blood stasis syndrome was higher that of sham operation group and health control group. After BMSCs transplant there was statistical difference of rats cardiac tissues between each groups (P＜0.05). The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). Every part of no BMSCs transplant of MMP-9 had statistical differences between each group. Cardiac blood stasis syndrome group was the highest, sham operation group was the next one and health control group was the last one. After BMSCs transplant, the cardiac blood stasis syndrome group was lower than that of health control group and sham operation group. The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). Every part of no BMSCs transplant of p-Akt had no statistical differences between each group.After BMSCs transplant, the cardiac blood stasis syndrome group was lower than that of health control group and sham operation group, the expression of transplant group's was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05).
Every part of no BMSCs transplant of p-p38 had no statistical differences between each group. There was no difference between each group's rats cardiac tissues after BMSCs transplant. The transplant group's expression was higher than that no BMSCs transplant of cardiac blood stasis in the same group(P＜0.05). The expression of GATA-4 mRNA,MMP-9 and p-p38 was positive correlation. The incidence coefficient were 0.49and 0.605 while there was no obvious associativity of MMP-2、p-Akt.
(3)Apr-YTF interfered in BMSCs transplant of cardiac blood stasis of Desmin-Brdu, the (rhG-CSF)+BMSCs group's positive cell counting were higher than that of NS+BMSCs group. There was no difference between (rhG-CSF)+BMSCs group and NS+BMSCs group. Through MHC-Brdu antibody dyeing, (rhG-CSF)+BMSCs and (apr-YTF)+BMSCs group's positive cell counting were higher than that of NS+BMSCs group. The semi-quantitative IOD of GATA-4 mRNA, NS+BMSCs group as well as (rhG-CSF)+BMSCs and (apr-YTF)+BMSCs group were higher than that of NS+DMEM group;while the (apr-YTF)+BMSCs group was higher than that of NS+BMSCs.
(4)Apr-YTF interfered in BMSCs transplant of cardiac blood stasis of MMP-2 and MMP-9, ELISA test showed that the MMP-2 (apr-YTF)+BMSCs group's expression was lower than that of NS+BMSCs group and NS+BMSCs. There was no difference between each group of MMP-9. Through p-Akt Western-Blot, (apr-YTF)+BMSCs's expression was higher than that of in NS+DMEM group NS+BMSCs group as well as (rhG-CSF)+BMSCs group. The p-p38 (apr-YTF)+BMSCs group's expression was higher than that of NS+DMEM group. There was no difference between each other group.
(5)After the induction of BMSCs simulated the environment about AMI cardiac blood stasis syndrome in vitro, there was no MHC positive cell been detected in the normal control group and DMEM group, while the weakly positive immunocomplex had been detected in the cardiac blood stasis syndrome group, apr-YTF group, cardiac blood stasis syndrome+apr-YTF in the Desmin and MHC immunohistochemistry dyeing. The IOD of Desmin expressionin the cardiac blood stasis syndrome group and cardiac blood stasis syndrome+(apr-YTF)group were higher than in the apr-YTF group, but there was no statistical difference between the former two ones。The GATA-4 mRNA IOD in cardiac blood stasis syndrome group and cardiac blood stasis syndrome+(apr-YTF)group had statistical differences compared with the normal control group.
(6)The MMP-2's mass concentration in the cell culture of simulated the environment of cardiac blood stasis syndrome+apr-YTF group was the highest which had the statistical difference compared with the normal control group DMEM and apr-YTF group. The mass concentration of MMP-9 in the two cardiac blood stasis syndrome group, and cardiac blood stasis syndrome+apr-YTF group were higher than in the DMEM and apr-YTF group.After 28 days of BMSCs' induction, the p-Akt in the signal transduction path was much higher in the cardiac blood stasis syndrome+apr-YTF group than that in the DMEM, apr-YTF and cardiac blood stasis syndrome group. The p-p38's expression in each group had no statistical difference.
Conclusion:
(1)The microenvironment of myocardial infarction cardiac blood stasis syndrome's cardiac cell can promote the BMSCs cardiac cell cytodifferentiation. Apr-YTF can promote it.
(2)The microenvironment of myocardial infarction cardiac blood stasis syndrome's cardiac cell can increase the expression of MMP-2 and MMP-9, promote the BMSCs migration and differentiation. After BMSCs transplant cardiac blood stasis syndrome's cardiac cell microenvironment's content expression lower and thus mitigate the tissure degradation cicatrization and control the deterioration of MI. Apr-YTF can lower the MMP's expression of AMI cardiac blood stasis syndrome in cardian tissure.
(3)PI3K signal transduction path in cardiac blood stasis syndrome's cardiac cell microenvironment has been activated. After BMSCs transplant cardiac blood stasis syndrome's cardiac cell microenvironment's cardiac cell differentiation related to the activation of PI3K and p38 signal transduction. Apr-YTF can promote the BMSCs transplant into cardiac cell differentiation and the activation of PI3K and p38 signal transduction.
(4)Simulated the microenvironment about cardiac blood stasis syndrome in vitro can induce the BMSCs cardiac cell differentiation. Apr-YTF can induce the BMSCs cardiac cell differentiation in vitro.
(5)Simulated the microenvironment about cardiac blood stasis syndrome in vitro, MMP-2 and MMP-9 can promote the BMSCs cardiac cell differentiation. BMSCs differentiation to cardiac cell and apr-YTF's interferences has related to the activation of PI3K signal transduction path.
引文
[1]http://www.nebi.nlm.nih.gov/sltes/entrez.ZOIO.1.26.
[2]DayanF，MazureNM，Brahimi-HornMC.etal.ADialogue between the Hypoxia-Indueible Factor and the Tumor Microenvironment[J」 .CaneerMieroenviron.2008，1(1):53-6.
[3]MaWW,AdjeiAA.Novel Agentson the Horizon for Cancer The raPy[Jl.CACaneer JC1in， 2009，59(4):111-137.
[4]TzukermanM，RosenbergT，Reiterl， etal.The Influence of a Human Embryonic stemCe11 DerivedMieroen vironmenton TargetingofHumanSolidTumorXenografts[J」.CancerRes，200，66(7):3792-3801.
[5]蒋雯，林国生，李庚山，等.兔骨髓间充质干细胞自体移植对缺血，二肌心功能的影响〔J].临床心血管病杂志，2004，20(10):605-608.
[6]高伟良.细胞信息网络[J].免疫学杂志，1997，13(4):211-213.
[7]http://trend.enki.net/trendshow.php?searehword=%ES%BE%AE%E7%SE%AF%ES%AZ%83&tjTyPe=&databasetj=define-search.2010.3.19.
[8]宋修竹，顾玉东，胡韶楠.神经再生室中神经再生过程和微环境的研究汇Jl.中华手外科杂志， 2001，17(1):39-40.
[9]王检芳，南雪，尉承泽，等.丙烯醇致肝损伤微环境定向诱导骨髓干细胞向肝细胞分化[J].中华肝脏病杂志，2005，23(4):274-277.
[10]王磊，徐坤.果袋材质对番茄果实微环境及其生长发育的影响[J]中国蔬菜， 2006，1:15-17.
[11]魏晓芳，丁西明，刘会洲.PH诱导牛血清白蛋白芳香氨基酸残基微环境变化的光谱分析[J].光谱学与光谱分析， 2000，20(4):556-559.
[12]胡多沙，曹亚.肿瘤微环境中的免疫效应抑制[1].国际病理科学与临床杂志， 2007，27(2):108-111.
[13]刘芳.缺氧微环境与实体瘤[J].现代诊断与治疗，20的， 18(1):36-39.
[14]http://trend.cnki.net/more.php?searchword=%ES%BE%AE%E7%SE%AF%ES%AZ%83&type=h15tory.2010.3.19.
[15]http://trend.cnki.net八rendshow.php?searehword二%ES%Bg%BZ%E7%BB%86%ES%83%gE%ES%BE%AE%E7%SE%AF%ES%AZ%83.2010.3.19.
[16]http://www.google.eom/trends?q=mieroenvironment&etab=0&geo=all&date=all.2010.3.19.
[17]http://www.google.en/trends?q=niche&ctab=0&geo=all&date=all.2010.3.19.
[l8]吴以岭主编.络病理论科学求证[Ml.北京:科学技术出版社， 2007:20-25.
[19]常富业，王永炎.络病辫证浅析[J].北京中医药大学学报，2003，26(6):9.
[20]李维革，韩宁.从络病理论探讨中风病微观病机[J].辽宁中医药大学学报， 2006，8(6):18.
[21]郑幸凡，陈刚.络脉的分布规律探讨[J].中医药信息， 2003，20(3):12.
[22]吴以岭.中医络病学说与三维立体网络系统[J].中医杂志，2003，44(6):407-409.
[23]翁维良.冠心病血癖证的研究进展[J].中医杂志，1987，25(6):464.
[24]郭志华.冠心病心绞痛2432例中医辫证分型综合统计分析[J].湖南中医杂志， 1998，14(2):7-8.
[25]衷敬柏，董绍英，王阶，等.2689例冠心病心纹痛证候要素的文献统计分析[J].中医药信息学，2006，13(5):100-101.
[26]周华兰，魏宗德.血检前状态与冠心病的研究[J].医学综述，2004，10(10):604-605.
[27]刘莉.凝血酶原片段F1+2在血检前状态研究及冠心病诊断防治中的意义[J].国外医学输血及血液学分册， 1994，17(5):286.
[28]马承恩，王可富，徐冬玲，等.急性冠状动脉综合征患者血液凝固性加强[J]，中国动脉硬化杂志，2006，14(5):438-440.
[29]郑春霞，刘志红.血管内皮细胞损伤及其检测[J].肾脏病与透析肾移植杂志， 2007，16(1):64-69.
[30]章宏祥，段宝祥.血浆血栓调节蛋白的含量与冠心病[J].医学综述，2005，11(3):218-220.
[31]B1akeGL，Ridker PM.Novele linical marker sofvaseu larwallin flame-mation J1.C1reRes，2001，89:763-771.
[32]阴彦龙，王仰坤.炎症因子与冠心病[J].海南医学，2007，18(6):125-127.
[33]王阶，杨保林，姜燕，等.冠心病血疥证相关基因研究[J].中日友好医院学报， 2005，19(5):314-316.
[34]毛以林，袁肇凯，黄献平，等.冠心病血癖证与血管紧张素转换酶基因多态性的相关性研究〔J].中国中西医结合杂志，2004，24(9):776-780.
[35]吴红金，马增春，高月，等.蛋白质组学技术对冠心病血疥证相关蛋白的研究[J].中西医结合心脑血管病杂志，2005，3(3):189-191.
[36]http://jnzsgbyybdxgy.blog.163.com/blog/static/130438043200991761318102/.2010.3.26.
[37]WangJS，shamTimD，GalipeauJ，etal.Marrow stromaeells for eellularear diomyopathy:feasibi1it yandpotentiale inicaladvantages[J].JThoraeCardiovascsurg，2000，120(5):999-1005.1381WangJS，ShamTimD，ChedrawyE，etal.Thecoronarydeliveryofmarrowstromaleellsformyocardialregeneration:PathoPhysiologieandtheraPeuticimPlications39]Wojakowski JThoracCardiovaseSurg，2001，122(4):699-705.TenderaM.Mobilizationofbonemarrow-derivedProgenitor[J]W，ee115inaeutesyndromes[J].FoliaHistoehemCytobiol，2005，43(4):229-232.[40〕AzharM，SchultzJE，GruPpeardiovaseulardeveloPment Rev，2003，14(5):391-407.I，et and a1.Transforminggrowthfaetorbetain funetion〔J].CytoltineGrowthFaetor
[41]L1nkeA，MullerP，NuxzynskaD，etal.Stem cell sin the dogheartareself-renewing，e1onogenie，andmulti Potentandregenerateinfraetedmyocardium，imProvingeardiaefunetion[J].PNAS，2005，102(25):8966-8971.
[42]HengB，HaiderH，SimEK，etal.Strategiesfordireetingthediffer-entiationof5tem cell sin to thecardiomyogeniclineageinvitro〔J]CardiovasRes，2004，62(1):34-42.
[43]KuangD，ZhaoX，XiaoG，etal.Stemeellfaetor/c-kitsignaling mediated cardia e5temeellmigration viaae tivation of P38MAPK[J].BasieResearehinCardiology，2008，103(3):265-273.
[44]OrlieD，KajsttuaJ，ChimentiS，etal.Bonefnarrow cell sregeneratein faretedmyoeardium[J].Nature，2001，410(6829):701-705.
[45]LiT，HayashiM，ItoH，etal.Regeneration of infareted myoeardium by intramy-oeardialim Plantation of exvivotrans for minggrowthfactor-3PreProgra皿edbonemarr0w5temcells[J」.Circulation，2005，111(19):2438-2445.
[46]SchenkS，MaIN，FinanA，etal.Monocy teehemo taeticprotein-315amyoear dialme Senchyma 15temcellholningfaetor[J].StemCells，2007，25(1):245-51.
[47]沈丰强，王爱玲，梁有峰，等.阿托伐他汀联合骨髓间充质干细胞移植治疗大鼠急性，。肌梗死[J].安徽医科大学学报，2009，44(5):558-561.
[48]贾战生.干病细胞治疗基J出与临床〔M〕.北京:人民卫生出版社，2005:267-268.
[49]DennisJE，CharbordP.originand differentiation of humanand murine Stroma[J].StemCells，2002，20(3):205-214.
[50]王宇玫.不同心脏病患者骨髓间充质干细胞培养结果的比较[J].感染、炎症、修复，2007，8(4):217-220.
[51]L1eehtyKW.Human mesene by malstem cell sengra ftanddemon stratesite speeifie differentiation after inutero transplantationinsheep[J」.JNatMed，2000，6(11):1282-1286.
[52]Tomitas，NakataniT，FukuharaS，etal.Bonemarrow stromal Cells contractsynchr onously with eardiomyoey tesinacoeul turesystem[J].JThoracCardiovaseSurg，2002，50(8):321-324.
[53]orlieD，KajsturaJ，ChirnentiS，etal.Bonemarrow Cell sregenera teinfracted myocardium[J].JNature，2001，410:701-705.
[54]李进领，周崇臣，孙春霞.灯盏细辛对骨髓间充质干细胞经静脉移植后对缺血性脑血管病的影响[J].中华实用中西医杂志，2007，20(23):2025-2027.
[55]曾意荣，樊粤光，刘红，等.补肾活血中药对大鼠骨髓间充质干细胞体外增殖的影响[J].中国组织工程研究与临床康复，2008，12(8):1545-2547.
[56]宋清，张晓文，卢新政，等.参三七皂普Rgl预适应对5-氮胞普诱导大鼠骨髓间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复，20的，11(24):4693-4697.
[57]GradnerRL，BrookFA.Reflection sonthebiologyo fembr yonics temells〔J」.IntJDevBiol，1997，41:235-243.
[58]Be1tramiAP，UrbanekK，KajturaJ，etal.Evidenee that human card iacmyoey tesdlvide after myoeardialin farction[Jl.NEnglJMed，2001，344(23):1750-1757.
[59]QuainiF，UrbanekK，BeltramiAP，etal.Chimerismofthetransplantedheart[J].NEnglJMed，2002，346(1):5-15.
[60]KajsturaJ，LeriA，FinatoN，etal.Myocyte proliferation inend-stage card iaefa1lurein humans[J].JProcNatl AeadSeiUSA，1998，95:8801-8805.
[61]装雪涛主编..干细胞生物学[M].北京:科学出版社,2003:4-7.
[62]P1ttenger干细胞生物学[MJ.北京:科学出版社，20的:4-7.MF，MaekayAM，BeekSC，etal.Multil image Potentialo fadul thumanme5e nehyma 15 temeells[J].Seience.1999，284(2):143-147.
[63]卜殷中，陆国平，张大维.超敏。-反应蛋白、基质金属蛋白酶-9在急性心肌梗死中的临床意义[J]医学临床研究，2008，25(5):817-819.
[64]DabekJ，KulachA，Gasior2.Theroleofma trixmetallo Proteinase sinaeute coronary syndromes[Jl.EurJInternMed，2007，18(6):463-466.
[65]NomotoK，oguchis，Watanabe1.Involve mento finflanunationinaeu teeoronary syndromesassessedbylevelsofhighsensitivityC-reactiveProtein，matrixmetalloProteinase-9andsolubevaseular-celladhesionmolecule-1[J」.JCardiol2003，42(5):201-206.
[66]G1ansanteC，FiottiN，DiChiaraA，etal.In-hosp italoutcomea cutecoronary syndrome relation shiP with in-flammation of Patientswithandremodelingmarkers[J].JCardiovaseMed(Hagerstown)，2007，8(8):602-607.
[67]L1JS，LiuK，WangMH，eta1.Modu1ationeffeet of naomai tongongea tinase system aftereere bralisehemia/rePer fusioninrats[J].ChinesejournaloflntegrativeMedieine，2006，26(SuPPI):14-17.
[68]WhitmarshAJ，YangS-H，SunMSS.Roleof p38 and JNK mitogen-aetivated proteinkinase in the activation of ternaryeom plexfactors[J].MolCellBiol，1997，17:2360-2371.
[69]RobinsonMJ，CobbMH.Mitogen-aetivated protein kinase path ways[J].Currop in CellB101，1997，9:180-186.
[70]Nemotos，ShengZ，LinA.opposin geffeets of junkina seand p38 mitogen-aetivated Proteinkin asesoncar diomyocyteshy PertroPhy[ Jl.MolCellBiol，1998，18:3518-3526.
[71]Ka1imanP，VinalsF，TestarX，etal.PhosPhatidy linositol3-kinaseinhibitors bloek differentiation of skeletal muscle cells[J].JBioChemi.1996，32(271):19146-19151.
[72]MenaseheP，HagegeAA， VilquinJT，etal.Autologouss keletal my oblasttrans-plantation for severe Postin farction left ventrieulardysfunetion[J」 .JAmCollCardiol，2003，41(7):1078-1083.
[73]DevineSM，HoffmanR.Role of mesenehymal stemeell sinhematopoietie stemeell trans plantation[Jl.CurropinHematol.2000，7(6):358-363.[74〕胡兵，安红梅，沈克，等.中医肾理论与干细胞研究[JJ.中国组织工程研究与临床康康复， 2008，12(3):598-601.
[75]郑景辉，李勇华，袁肇凯.骨髓间充质细胞中医药干预研究的现状及其前景和策略[J].辽宁中医药大学学报，2009，11(s):35-37.[76」汪朝晖，洗绍祥，杨忠奇，等.人参总皂贰诱导骨髓间充质干细胞分化为，二肌样细胞的实验研究[J〕.广州中医药大学学报，2006，23(2):100-103.
[77]王康锋，张洪斌，张立娟.中医肾精理论与神经干细胞关系探讨[J].新中医，2005，37(12):76-77.
[78]张进，徐志伟，陈群，等.干细胞与中医基础理论中的先天之精学说[J].中国组织工程研究临床康复，2006，10(7):189-192.
[79]FriedensteinAJ， PetrakovaKV， KurolesovaAl，etal.HeterotoPic of bonemarrow:Analy sis of preeursoreells forost eogenieand hema to Poietie tissues[J].TransPlantation，1968，6(2):230-247.
[80]马力，刘大军，李德天，等.不同分离方法及培养条件对兔骨髓间充质干细胞生长增殖及生物学特性的影响[J].中国组织工程研究与临床康复，2008，12(38):7401-7406.
[81]卢新政，张晓文，黄峻，等.大鼠骨髓间充质干细胞培养方法的改良[J].中国心血管杂志2004，9(1):48-51.
[82]YoshimuraH，MunetaT，NimuraA，eta1.Comparisonofratmesenehyma1stemcel1sderivedfrombonemarrow，synovium，Periosteum，adiPosetissueandmusele[J」.Ce11TissueRes，2007，327(3):449一462.
[83]HughesGC，PostMJ，SimonsM，etal.TranslationalPhysiology:Poreinemodelsofhumaneoronaryarterydisease:imPlicationsforPreelinicaltrialsoftherapeutieangigenesisfJ}.ApplPhysiol，2003，94:1689一1701.
[84]PaganiFD，SimonianHD，ZawadzkaA，etal.AutologousSkeletalMyoblastsTransPlantedtoisehemiadamagedmyocardiuminHumans〔J].JournaloftheAmericanCollegeofCardiology，2003，(41):879一888.
[85]黄国倩，刘虹，舒先红，等.超声心动图评价粒细胞集落刺激因子动员自身骨髓干细胞对心肌梗死后左心室重构及心功能的影响[J1.中华超声影像学杂志，2004，13(11):848-852.
[86]和岚，蒋文跃，毛腾敏.注射肾上腺素模拟”气滞”复制急、慢性血疥模型初探[J].中国中西医结合杂志，2004，24(3):244一246.
[87]ZhangH，XuC，LiangL，etal.Participation of bonemarrow derivedme senchymal stemcell sine or nealepi the lial wound healing[J].Invest.ophthalmol.Vis.Sei，2008，49:6071.
[88]李玉玲，唐俊明，潘国栋，等.DAPI标记的骨髓间充质干细胞体内外示踪实验研究[J].员卜阳医学院学报，2005，24(5):257-260.
[89]刘伯轩，刘师伟，王建华，等.DAPI标记的骨髓间充质干细胞体内外示踪实验研究[J]中国药物与临床，2009，9(8):703一705.
[90]orieD，KajsturaJ，ChimentiS，etal.Bonemarrow cell sregener ateinfraeted myoeardium[J].Natrue，2001，410:701-705.
[91]牛丽丽，祝善俊，张树林，等.同种异体骨髓间充质干细胞在大鼠心脏的迁移及分化特点[J].中华心血管病杂志，2004，32(增刊):186-189.
[92]GrossJ，LapiereCM.Collage nolyticae tivityi namphibian tissues:atissuecul tureassay[J].proeNatl AeadSciuSA.1962，15(48):1014-22.
[93]李建生，刘敬霞，孙捷，等.大黄普元对骨髓间充质干细胞移植脑缺血大鼠基质金属蛋白酶的影响[J].中西医结合学报，2008，6(s):510一816.
[94]ChengS，LovettDH.GelatinaseA(MMP一2)15necessaryandsufficientforrenaltubulareellepithelialmesenehymaltransformation[J].AmJPathol，2003，162(6):1937-1949.
[95]郑丽芳，张红，张小乔.GF一pl对骨髓间充质干细胞迁移力及Snail表达的影响[J].细胞生物学杂志，2005，30:222一228.
[96]SpinaleFG，CokerML，KrombaehSR，etal.Matrixmetal loproteinaseinhibition during the develoPment of eongestive heart failure:effectsonleftventrieulardimensionsandfunction[J].CircRes，1999，85:364一376.
[97]SpinaleFG，CokerML，HeungLJ，etal.Amatrixme talloproteinase induction/aetivation system exist sin the human left ventrieular myoear diumandlau Pregulate dinheartfailure[J].Circulation，2000，102:1944-1949.
[98]NagayaN，KangavaK，etal.Transplantationofmesenehymalstemeellsimprovescardicfunctioninaratmodelofdilatedcardiomyopathy[J].Circulation2005，112(8):1128-1135.
[99]雷警瑜，王卫淑，姚巍.间充质干细胞移植对，。肌基质金属蛋白酶一9及抑制因子-1表达的影响〔J].临床，“血管病杂志，2009，25(6):453一456
[100]Ka1imanP，VinalsF，TestarX，etal.PhosPhatidylinositol-3kinaseinhibitorsb1oekdifferentiationofskeletalmuscleeells[J」.JBioChemi，1996，32(271):19146-19151.
[101]KrasilnikovMA.PhosPhatidy linosito-3kinase dePendent Pathways:the roleineon trolofeell growth，survival，andmalignanttransformation[J」.Bioehemistry，2000，65(1):59-671.
[102]JaiswalRK，JaiswalN，BruderSP，etallAdul thumanme senehymal stemeell di-fferrentiationto the osteogenicoradiPogeniclineage15regulatedbym1toZgenZactivatedproteinkinase[J].JBiolChem，2000，275(13):9645一9652.
[103]王劲，郭朝华，罗成基，等.成肌细胞增殖n分化的信号转导研究[J]1生命的化学，2002，22(1):18-20.
[104]王劲，罗成基，郭朝华，等.p38及P工3K的激活在工GF21促进BMSCs成肌分化中的作用[J].第三军医大学学报，2003，25(13):1137-1139.
[105]Hami doucheZ，HaLE，VaudinP，etal.FHLZ media tesdexamethasone-indueed mesenehy maleell differentiation intoo steoblastsbyaetivatingwnt/p-catenin Signaling-dependent RunxZex pression[J].FASEBJ，2008，22:3813一3822.
[106]Boeelli-TyndallC，BracciL，SchaerenS，eta1.Human bone marrowme senehyma stem cells and chondrocytespromoteand/orsuPPresstheinvitroProliferationoflymphoeytesstimulatedbyinterleukinsZ，7and15[Jl.AnnRheumDis，2009，68:1352-1359.
[107]MatsushitaK，WuY，okamotoy，etal.local reninang iotensin expression regulate shumanme senehymal stemcell differentiation toad ipocytes[J].Hypertension，200648:1095-1102.
[108]Aurichl，MuellerLP，AurichH，etderivedfrom human mesene by mala1.Functional integration The Patocytes Stem cell sintomouselivers[J].Gut，2007，56:405-415.
[109]MartinezC，HoffnannTJ，MarinoR，etal Human bone marrowme senehymal stromaleell sex Press then euralgang liosideGDZ:ofBMSCs[J].Blood，2007，109:anovelSurfacemarkerfortheidentification4245-4248.
[110]HakunoD，FukudaK.Bonemarrow-derivedregeneratedeardiomyocytes(CMGcells)expressfunetionaladrenergicandmusearinicrecePtor[J].Cireulation2002，105(3):380-386.
[111]StrauerBE，BrehmM，ZeusT，etal.Repairofin fracted myocardiumby auto logousin tracoronary mononuelearbonemarrowcelltransplantationinhumans[J].C1rculation，2002，106(15):1913一1918.
[112]Tomitas，MiekleDA，WeiselRD，etal.Improvedheartfunetion with my ogenesis and angiogenesisafter auto logouspo reinemarrow stromaleelltran splan tation[J].JThoracCardiovaseSurg，2002，123:1132-1140.
[113]秦伯未.谦斋医学讲稿〔M〕.上海科学技术出版社，新1版，1978:2此.
[114]袁肇凯，周泽泉.养心通脉片治疗冠心病心绞痛的临床研究[J].中药新药与临床药理，1998，9(1):19一21.
[115]史学军，闰文浩.养心通脉方治疗冠心病51例疗效观察[J].中国医刊，1999，34(7):50-51.
[116]衰肇凯，曹金枝，黄献平，等.养心通脉液对心肌缺血再灌注损伤大鼠冠状微循环的影响[J」.中西医结合心脑血管病杂志，2004，2(12):715-717.
[117]王东生，袁肇凯，黄献平，等.养心通脉片对动脉粥样硬化大鼠“疾癖”病理细胞凋亡的影响〔J〕.中医杂志，2004，45(12):937-940.
[118]卢芳国，衰肇凯，黄献平，等.养心通脉片对胰岛素抵抗综合征外周血单核细胞C-myc和PDGF一AmRNA的影响〔J].中西医结合心脑血管病杂志，2003，1(8):442-444.
[119]莫莉，黄献平，袁肇凯，等.养心通脉有效部位方对鸡胚绒毛尿囊膜血管新生的影响[J〕中药新药与临床药理，2009，20(4):315一319.
[120]哀肇凯，莫莉，黄献平，等.养心通脉有效部位方对心肌缺血大鼠心血管内皮细胞血管生成的影响[J]一。血管康复医学杂志，2009，18(2):180一184.
[121]ColterDC，ClassR，DIGirolamoCM，eta1.Rapidexpansionofreeye1ingstemce11sinculturesofPlastie一adherent cells from human bone marrow[J].ProcNatlAcadSeiUSA，2000，97(7):3213-3218.
[122]郭艳，史大卓，陈可冀.中医药结合干细胞移植治疗心肌损伤性疾病的前景[J].新中医[J].2005，37(5):88-89.
[123]oswaldJ，BoxbergerS，JorgensenB，etal.Mesenchymals temcellseanbe different iatedinto and othelial cell sinvitro[Jl.StemCells，2004，22:377-384.
[124]ShengXG，SongH，FengJZ，etal.VEGF-exPressingbone marrowme senchymal stemeell strans Plantationim Provedheart function of myoeardialinfare trabbits[J]South China Journalo fear diology，2006，6(1):14-17.
[125]洗绍祥，杨忠奇，汪朝晖，等.黄茂甲普体外诱导骨髓间充质千细胞分化为心肌样细胞的实验研究[J]广州中医药大学学报，2007，24(1):37-40.
[126]宋清，张晓文，卢新政，等.参三七皂普Rgl预适应对5-氮胞普诱导大鼠骨髓间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复，2007，11(24):4693-4697.
[127]章亭，谭允育，潘彦舒.四物汤对红细胞免疫及骨髓干细胞增殖能加勺影响[J].北京中医药大学学报，2000，23(1):36-38.
[128]王春燕，曾和平，周亚东，等.补阳还五汤乙醇、氯仿提取物挥发性成分及调控鼠骨髓间充质干细胞活性研究[J]华南师范大学学报(自然科学版)2007，3:88-92.
[129]黄明明，崔晓瑞，邵雪喜.虫草类的免疫药理研究工.人工发酵的虫草菌的免疫抑制作用的观察[J].中华器官移植杂志，1984，5(2):58.
[130]曹丰，贾国良，牛丽丽，等.移植时机对骨髓间充质干细胞修复梗死心肌的影响[J].第四军医大学学报，2005，26(1):22-27.
[131]SuzukiK，SmolenskiRT，JayakumarJ，etal.Heat shoek treatment enhanees graft cell surviva linske letal my oblast transplantationtotheheart[J].Circulation，2000，102(19SupP13):111216-111221.
[132]GnecchiM，HeH，NoiseuxN，etal.EvideneesuPportingparaerinehypothesisforAkt-modifiedme senehymal stemeell-media tedear diaeProtection and funetional improvement[J].FASEBJ，2006，20(6):661-669.
[133]Jiangs，HaiderHKh，IdrisNM，etal.Supportiveinteraetionbetweeneellsurvival5ignaling and angioeom Petent faetor senhaneesdonor cell survival and Promotesang iomyogenesis for eardiae repair[J].CireRes.ZO06，99(7):776-784.
[134]GneeehiM，HeH，NoiseuxN，etal.Evideneesup Porting paraer inehypo thesis for Akt-modifiedmesenchymalstemcell-mediatedeardiac Proteetionand functional improvement [J].FASEBJ，2006，20(6):661-669
[135]TangYL，TangY，ZhangC，etal.Improved graft mesenehy malstem cell surviva linsehemi cheart with ahypoxia-regulated Hemeoxygenase-1veetor[J].JAmColl Cardiol，2005，46(7):1339-1350.
[136]JoJl，NagayaN，MiyaharaY，etal.Transplantation of geneti cell yengineered mesenehymalin faretion:Stem cell sim Provescardiaefunetionbenefitofanovelnonviralvector[J].inrats with myoear dialCationized DextranTISsueEng，2007，13(2):313-322.
[137]LIW，MaN，OngLL，etal.Bcl-2engine ered BMSC sinhibitedaPoPtosis and imProved heart function[J].StemCells.2007，25(8):2118-2127.
[138]BartunekJ，CroissantJD，WijnsW，etal.Pretreatfnen to fadult bone marrowme sene by malstem cell swithcar diomyogenie growthfactorsandrePairofthechronieallyinfaretedmyocardium[J].AmJPhysiolHeartCirePhysiol，2007，292(2):1095-1104.
[139]Voytik-HarbinSL.Three-dimensional extra cell ularmatrixsub strates fore ellculture[Jl.Methods Cell Biol，2001，63:561-81.
[140]PaulD，SamuelSM，MaulikN.Mesen chymal stemeell:present chall enges and pros Pective cellular cardiomyo PlastyaP Proaches for myocardia1regeneration[J].AntioxidRedoxSignal，2009，11(8):1841-1855.
[141]ZhangY，ChuY，ShenW，etal.Effectofs-azaey tidine induction durationond differentiation of human first-trimes terfetalmesenehymalstemeellstowardscardiomyocyte-likeeells[J].InteraetCardiovascThoracSurg，2009，9(6):943-946.
[142]HuangJL，YangSX.Safetyofumbiliealeordblood-derivedmesenehymalstemeells(BMSCs)followings-azaser ineinductionandinhibitionofhumancardiaemyoeyteapoptosisbyBMSCs[J].SaudiMedJ，2009，30(9):1144-1149.
[143]JYeNS，ZhangRL，ZhaoYF，etal.Effectofs-azaeytidineon the proteinex Pression of Porcine bone marrowme senehymal stem cell sinvitro[J].Genomies Proteomics Bin for maties，2006，4(1):18-25.
[144]Moscosol，CentenoA，LoPezE，etal.Differentiation”invitro”ofinortalizedPoreinemesenehymalstemeellsintocardiomyoeytestransplantation[J].Transplant ation Proeeedings，2005，37(1):481-482.Primaryandforcell
[145]RameshwarP，QiuH，VatnerSF.Stem cell sincardiae repair inaninflam atory meroen vironment[J』.Minerva Cardioangiol，2010，58(1):127-146.
[146]李艳芬，孙兰军，赵英强，等.复方丹参滴丸干预骨髓间充质千细胞移植后，。肌再生的实验研究[J].中国中医药现代远程教育，2009，8(9):81-83.
[147]南亚均，李庆雯，王宇春，等.BMSc联合淫羊蓉普预处理的EPCS移植对AMI大鼠心功能的影响[J].山西中医，2009，25(2):44-46.
[148]HayloekDN，NilssonSK.Stem cell regulation by the hematopoie tiestem cellniehe[J」.Cellcycle，2005，4(10):1353-1355.
[149]NilssonSK，SinunonsPJ.Transplan table stem cells:hometo speeificniches[J].CurrOPin Hematol，2004，11(2):102-106.
[150]WitzIP.Yin-Yangactivities andvieiouse yelesin the tumor mieroen vironment[J].CaneerRes，2008，68(1):9-14.
[151]HengBC，HaiderHK，SimEK，etal. Strate gies fordirecting the differentiation of stem cell sin to the cardiomyogeni clineagein vitro[Jl.CardiovaseRes，2004，62(1):34-42.
[152]张金红，耿朝晖，段江燕，等.MTT比色法在贴壁细胞药物敏感型测定中的应用[J].南开大学学报(自然科学版)，1996，29(4):109.
[153]MakinoS，Fukudak，Miyoshi5.etal.Cardiomyo eyteseanbegener ated from marrow strofnal cell sinvitro[Jl.J-ClinInvest，1999，103(5):697-705.
[154]YoonJ，MinBG，KimYH，etal.Differentiation，engraftment and funetion aleffects of pre-treatedme senehymalstem cell sinaratmyoeardialinfaretmodel[Jl.AetaCardiol，2005，60(3):277-284.
[155]ShimWS，JiangS，WongP，etal.Exvivodifferentiationofhumanadu1tbonemarrow5temcellsintoeardiomyoeyte-likeeells[J].BioehemBioPhysResCo朋un，2004，324(2):481-488.
[156]肖诗亮，郑日善.不同方法诱导的骨髓间充质干细胞移植修复大鼠梗死心肌〔J].华中科技大学学报(医学版)2007，36(3):339-342.
[157]常静，雷寒，陈建斌，等.大鼠心肌细胞诱导人骨髓问充质千细胞向，。肌样细胞分化的研究[J].重庆医科大学学报，2007，32(9):907-910.
[158]Ba11SQ，Shuttle worth AC，Kielty CM，etal.Direct cell contactin fluenees bonemarrowmese neymalstem cell fate[J].IntJBioehemCellBio，2004，36(4):714-727.
[159]RangappaS Ent wistle JWWeehslerA Setal.Cardiomyoeyte media tedeontaetprograms human mesenchymal stem cells to express eardiogenie phenotype[J].J Thorac Cardio vasc Surg，2003，126(1):124-132.
[160]XuM，WaniM，DaiYS，etal.Differentiation of bone marrow stromal cells into the eardiae Phenoty Perequires intercellular communieation with myocytes[J].Circulation，2004，110(17):2658-2665.
[161]YuanYC，henLF，ZhangSY，etal.Differentiation of mesenehy malstemcell sintoeardi of nyogeniceell sunder the induetion of myoeardia leellly sate[Jl.Chinese Journal of Cardiology，2005，33(2):170-173.
[162]汪朝晖，洗绍祥，杨忠奇，等.人参总皂戒诱导骨髓间充质干细胞分化为，。肌样细胞的实验研究[J].广州中医药大学学报，2006，23(2):100-103.
[163]李艳芬，孙兰军，赵英强，等.复方丹参滴丸干预骨髓间充质干细胞移植后心肌再生的实验研究[J].中国中医药现代远程教育，2009，9:81-83.
[164]陆长青，冉黎，张全波，等.丹参诱导骨髓间充质干细胞向神经元样细胞分化及相关基因的表达[J].中国组织工程研究与临床康复，2005，12(47):9363-9366.
[165]LueianoCA，AnastasiosPS，KarlHS，etal.Cardiae repair with intramyo cardialin jeetiono fall ogeneic mesenehymal stemeell saftermyoear dialin faretion[J].PNAS，2005，102(32):11474-11479.
[166]XuM，WaniM，DaiYS，etal.Differentiation of bonemarrow stromal cellsinto the cardiae phenoty Perequires in tereellular conununication with myoeytes[J].C1rculation，2004，110(17):2658-2665.
[167]曹寅，潘呈.三维细胞培养技术研究进展[J].安徽医药，2006，20(3):221-222.
[168]DoueetC，Ernoul，ZhangY，etal.PlateletlysatesPromotemesenehymalstemeellexPansion:asafetysubstitutefor2005，animalserumincell-basedtheraPyaPplications[J].JCellPhysiol，205(2):228-236.
[169]PittengerMF，MaekayAM，BeckSC，etal.Multi-lineage protential of adult humanmes enehymal stemcells[J].seienee，1999，284:143-147.
[1]Colter DC,Class R, DiGirolamo CM, et al.Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow[J].Proc Natl Acad Sci USA,2000,97(7):3213-3218.
[2]郭艳,史大卓,陈可冀.中医药结合干细胞移植治疗心肌损伤性疾病的前景[J].新中医,2005,37(5)：88-89.
[3]Oswald J,Boxberger S, Jorgensen B, et al.Mesenchymal stem cells can be differentiated into endothelial cells in vitro[J]. Stem Cells, 2004,22:377-384.
[4]赵春华.干细胞原理、技术与临床[M].北京化学工业出版社,2006：6290-6291.
[5]Jiang W,Ma A, Wang T, et al. Homing and differentiation of mesenchymal stem cells delivered intravenously to ischemic myocardium in vivo:a time-series study[J]. Pflugers Arch,2006,453(1):43-52.
[6]Penna C, Raimondo S, Ronchi G, et al. Early homing of adult mesenchymal stem cells in normal and infarcted isolated beating hearts [J]. J Cell Mol Med,2008,12 (2):507-521.
[7]Doucet C,Ernou I,Zhang Y,Llense JR,et al.Platelet lysates promote mesenchymal stem cell expansion:a safety substitute for animal serum in cell-based therapy applications[J]. J Cell Physiol,2005,205(2): 228-236.
[8]Pittenger MF,Mackay AM, Beck SC,et al.Multi-lineage protential of adult human mesenchymal stem cells[J]. science,1999,284:143-147.
[9]肖诗亮,郑日善.不同方法诱导的骨髓间充质干细胞移植修复大鼠梗死心肌[J].华中科技大学学报(医学版)2007,36(3)：339-342.
[10]常静,雷寒,陈建斌,等.大鼠心肌细胞诱导人骨髓问充质干细胞向心肌样细胞分化的研究[J].重庆医科大学学报,2007,32(9)：907-910.
[11]李进领,周崇臣,孙春霞.灯盏细辛对骨髓间充质干细胞经静脉移植后对缺血性脑血管病的影响[J].中华实用中西医杂志,2007,20(23)：2025-2027.
[12]曾意荣,樊粤光,刘红,等.补肾活血中药对大鼠骨髓间充质干细胞体外增殖的影响[J].中药新药与临床药理,2007,18(3)：93-96.
[13]宋清,张晓文,卢新政,等.参三七皂苷Rgl预适应对5-氮胞苷诱导大鼠骨髓 间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复,2007,11(24)：4693-4697．
[14]章亭,谭允育,潘彦舒.四物汤对红细胞免疫及骨髓干细胞增殖能力的影响[J].北京中医药大学学报,2000,23(1)：36-38.
[15]王春燕,曾和平,周亚东,等.补阳还五汤乙醇、氯仿提取物挥发性成分及调控鼠骨髓间充质干细胞活性研究[J].华南师范大学学报(自然科学版),2007,3：88-92.
[16]黄明明,崔晓瑞,邵雪喜.虫草类的免疫药理研究Ⅰ.人工发酵的虫草菌的免疫抑制作用的观察[J].中华器官移植杂志,1984,5(2)：58.
[17]商洪才,张伯礼.缺血性中风急性期治疗现状及中医药的前景和策略[J].天津中医药,2005,22(12)：523-525.
[18]赵春华.干细胞原理、技术与临床[M].北京化学工业出版社,2006：625-627.
[19]贾延,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[J].中华医学杂志,2002,82(19)：1337-1341.
[20]阎德文,李海燕,吴清平,等.GSB中药血清对rBMSCs定向诱导分化为成骨细胞的影响[J].中国骨质疏松杂志,2007,13(7)：480-485.
[21]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[J].广州中医药大学学报,2007,24(1)：37-40.
[2]DayanF，MazureNM，Brahimi-HornMC.etal.ADialogue between the Hypoxia-Indueible Factor and the Tumor Microenvironment[J」 .CaneerMieroenviron.2008，1(1):53-6.
[3]MaWW,AdjeiAA.Novel Agentson the Horizon for Cancer The raPy[Jl.CACaneer JC1in， 2009，59(4):111-137.
[4]TzukermanM，RosenbergT，Reiterl， etal.The Influence of a Human Embryonic stemCe11 DerivedMieroen vironmenton TargetingofHumanSolidTumorXenografts[J」.CancerRes，200，66(7):3792-3801.
[5]蒋雯，林国生，李庚山，等.兔骨髓间充质干细胞自体移植对缺血，二肌心功能的影响〔J].临床心血管病杂志，2004，20(10):605-608.
[6]高伟良.细胞信息网络[J].免疫学杂志，1997，13(4):211-213.
[7]http://trend.enki.net/trendshow.php?searehword=%ES%BE%AE%E7%SE%AF%ES%AZ%83&tjTyPe=&databasetj=define-search.2010.3.19.
[8]宋修竹，顾玉东，胡韶楠.神经再生室中神经再生过程和微环境的研究汇Jl.中华手外科杂志， 2001，17(1):39-40.
[9]王检芳，南雪，尉承泽，等.丙烯醇致肝损伤微环境定向诱导骨髓干细胞向肝细胞分化[J].中华肝脏病杂志，2005，23(4):274-277.
[10]王磊，徐坤.果袋材质对番茄果实微环境及其生长发育的影响[J]中国蔬菜， 2006，1:15-17.
[11]魏晓芳，丁西明，刘会洲.PH诱导牛血清白蛋白芳香氨基酸残基微环境变化的光谱分析[J].光谱学与光谱分析， 2000，20(4):556-559.
[12]胡多沙，曹亚.肿瘤微环境中的免疫效应抑制[1].国际病理科学与临床杂志， 2007，27(2):108-111.
[13]刘芳.缺氧微环境与实体瘤[J].现代诊断与治疗，20的， 18(1):36-39.
[14]http://trend.cnki.net/more.php?searchword=%ES%BE%AE%E7%SE%AF%ES%AZ%83&type=h15tory.2010.3.19.
[15]http://trend.cnki.net八rendshow.php?searehword二%ES%Bg%BZ%E7%BB%86%ES%83%gE%ES%BE%AE%E7%SE%AF%ES%AZ%83.2010.3.19.
[16]http://www.google.eom/trends?q=mieroenvironment&etab=0&geo=all&date=all.2010.3.19.
[17]http://www.google.en/trends?q=niche&ctab=0&geo=all&date=all.2010.3.19.
[l8]吴以岭主编.络病理论科学求证[Ml.北京:科学技术出版社， 2007:20-25.
[19]常富业，王永炎.络病辫证浅析[J].北京中医药大学学报，2003，26(6):9.
[20]李维革，韩宁.从络病理论探讨中风病微观病机[J].辽宁中医药大学学报， 2006，8(6):18.
[21]郑幸凡，陈刚.络脉的分布规律探讨[J].中医药信息， 2003，20(3):12.
[22]吴以岭.中医络病学说与三维立体网络系统[J].中医杂志，2003，44(6):407-409.
[23]翁维良.冠心病血癖证的研究进展[J].中医杂志，1987，25(6):464.
[24]郭志华.冠心病心绞痛2432例中医辫证分型综合统计分析[J].湖南中医杂志， 1998，14(2):7-8.
[25]衷敬柏，董绍英，王阶，等.2689例冠心病心纹痛证候要素的文献统计分析[J].中医药信息学，2006，13(5):100-101.
[26]周华兰，魏宗德.血检前状态与冠心病的研究[J].医学综述，2004，10(10):604-605.
[27]刘莉.凝血酶原片段F1+2在血检前状态研究及冠心病诊断防治中的意义[J].国外医学输血及血液学分册， 1994，17(5):286.
[28]马承恩，王可富，徐冬玲，等.急性冠状动脉综合征患者血液凝固性加强[J]，中国动脉硬化杂志，2006，14(5):438-440.
[29]郑春霞，刘志红.血管内皮细胞损伤及其检测[J].肾脏病与透析肾移植杂志， 2007，16(1):64-69.
[30]章宏祥，段宝祥.血浆血栓调节蛋白的含量与冠心病[J].医学综述，2005，11(3):218-220.
[31]B1akeGL，Ridker PM.Novele linical marker sofvaseu larwallin flame-mation J1.C1reRes，2001，89:763-771.
[32]阴彦龙，王仰坤.炎症因子与冠心病[J].海南医学，2007，18(6):125-127.
[33]王阶，杨保林，姜燕，等.冠心病血疥证相关基因研究[J].中日友好医院学报， 2005，19(5):314-316.
[34]毛以林，袁肇凯，黄献平，等.冠心病血癖证与血管紧张素转换酶基因多态性的相关性研究〔J].中国中西医结合杂志，2004，24(9):776-780.
[35]吴红金，马增春，高月，等.蛋白质组学技术对冠心病血疥证相关蛋白的研究[J].中西医结合心脑血管病杂志，2005，3(3):189-191.
[36]http://jnzsgbyybdxgy.blog.163.com/blog/static/130438043200991761318102/.2010.3.26.
[37]WangJS，shamTimD，GalipeauJ，etal.Marrow stromaeells for eellularear diomyopathy:feasibi1it yandpotentiale inicaladvantages[J].JThoraeCardiovascsurg，2000，120(5):999-1005.1381WangJS，ShamTimD，ChedrawyE，etal.Thecoronarydeliveryofmarrowstromaleellsformyocardialregeneration:PathoPhysiologieandtheraPeuticimPlications39]Wojakowski JThoracCardiovaseSurg，2001，122(4):699-705.TenderaM.Mobilizationofbonemarrow-derivedProgenitor[J]W，ee115inaeutesyndromes[J].FoliaHistoehemCytobiol，2005，43(4):229-232.[40〕AzharM，SchultzJE，GruPpeardiovaseulardeveloPment Rev，2003，14(5):391-407.I，et and a1.Transforminggrowthfaetorbetain funetion〔J].CytoltineGrowthFaetor
[41]L1nkeA，MullerP，NuxzynskaD，etal.Stem cell sin the dogheartareself-renewing，e1onogenie，andmulti Potentandregenerateinfraetedmyocardium，imProvingeardiaefunetion[J].PNAS，2005，102(25):8966-8971.
[42]HengB，HaiderH，SimEK，etal.Strategiesfordireetingthediffer-entiationof5tem cell sin to thecardiomyogeniclineageinvitro〔J]CardiovasRes，2004，62(1):34-42.
[43]KuangD，ZhaoX，XiaoG，etal.Stemeellfaetor/c-kitsignaling mediated cardia e5temeellmigration viaae tivation of P38MAPK[J].BasieResearehinCardiology，2008，103(3):265-273.
[44]OrlieD，KajsttuaJ，ChimentiS，etal.Bonefnarrow cell sregeneratein faretedmyoeardium[J].Nature，2001，410(6829):701-705.
[45]LiT，HayashiM，ItoH，etal.Regeneration of infareted myoeardium by intramy-oeardialim Plantation of exvivotrans for minggrowthfactor-3PreProgra皿edbonemarr0w5temcells[J」.Circulation，2005，111(19):2438-2445.
[46]SchenkS，MaIN，FinanA，etal.Monocy teehemo taeticprotein-315amyoear dialme Senchyma 15temcellholningfaetor[J].StemCells，2007，25(1):245-51.
[47]沈丰强，王爱玲，梁有峰，等.阿托伐他汀联合骨髓间充质干细胞移植治疗大鼠急性，。肌梗死[J].安徽医科大学学报，2009，44(5):558-561.
[48]贾战生.干病细胞治疗基J出与临床〔M〕.北京:人民卫生出版社，2005:267-268.
[49]DennisJE，CharbordP.originand differentiation of humanand murine Stroma[J].StemCells，2002，20(3):205-214.
[50]王宇玫.不同心脏病患者骨髓间充质干细胞培养结果的比较[J].感染、炎症、修复，2007，8(4):217-220.
[51]L1eehtyKW.Human mesene by malstem cell sengra ftanddemon stratesite speeifie differentiation after inutero transplantationinsheep[J」.JNatMed，2000，6(11):1282-1286.
[52]Tomitas，NakataniT，FukuharaS，etal.Bonemarrow stromal Cells contractsynchr onously with eardiomyoey tesinacoeul turesystem[J].JThoracCardiovaseSurg，2002，50(8):321-324.
[53]orlieD，KajsturaJ，ChirnentiS，etal.Bonemarrow Cell sregenera teinfracted myocardium[J].JNature，2001，410:701-705.
[54]李进领，周崇臣，孙春霞.灯盏细辛对骨髓间充质干细胞经静脉移植后对缺血性脑血管病的影响[J].中华实用中西医杂志，2007，20(23):2025-2027.
[55]曾意荣，樊粤光，刘红，等.补肾活血中药对大鼠骨髓间充质干细胞体外增殖的影响[J].中国组织工程研究与临床康复，2008，12(8):1545-2547.
[56]宋清，张晓文，卢新政，等.参三七皂普Rgl预适应对5-氮胞普诱导大鼠骨髓间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复，20的，11(24):4693-4697.
[57]GradnerRL，BrookFA.Reflection sonthebiologyo fembr yonics temells〔J」.IntJDevBiol，1997，41:235-243.
[58]Be1tramiAP，UrbanekK，KajturaJ，etal.Evidenee that human card iacmyoey tesdlvide after myoeardialin farction[Jl.NEnglJMed，2001，344(23):1750-1757.
[59]QuainiF，UrbanekK，BeltramiAP，etal.Chimerismofthetransplantedheart[J].NEnglJMed，2002，346(1):5-15.
[60]KajsturaJ，LeriA，FinatoN，etal.Myocyte proliferation inend-stage card iaefa1lurein humans[J].JProcNatl AeadSeiUSA，1998，95:8801-8805.
[61]装雪涛主编..干细胞生物学[M].北京:科学出版社,2003:4-7.
[62]P1ttenger干细胞生物学[MJ.北京:科学出版社，20的:4-7.MF，MaekayAM，BeekSC，etal.Multil image Potentialo fadul thumanme5e nehyma 15 temeells[J].Seience.1999，284(2):143-147.
[63]卜殷中，陆国平，张大维.超敏。-反应蛋白、基质金属蛋白酶-9在急性心肌梗死中的临床意义[J]医学临床研究，2008，25(5):817-819.
[64]DabekJ，KulachA，Gasior2.Theroleofma trixmetallo Proteinase sinaeute coronary syndromes[Jl.EurJInternMed，2007，18(6):463-466.
[65]NomotoK，oguchis，Watanabe1.Involve mento finflanunationinaeu teeoronary syndromesassessedbylevelsofhighsensitivityC-reactiveProtein，matrixmetalloProteinase-9andsolubevaseular-celladhesionmolecule-1[J」.JCardiol2003，42(5):201-206.
[66]G1ansanteC，FiottiN，DiChiaraA，etal.In-hosp italoutcomea cutecoronary syndrome relation shiP with in-flammation of Patientswithandremodelingmarkers[J].JCardiovaseMed(Hagerstown)，2007，8(8):602-607.
[67]L1JS，LiuK，WangMH，eta1.Modu1ationeffeet of naomai tongongea tinase system aftereere bralisehemia/rePer fusioninrats[J].ChinesejournaloflntegrativeMedieine，2006，26(SuPPI):14-17.
[68]WhitmarshAJ，YangS-H，SunMSS.Roleof p38 and JNK mitogen-aetivated proteinkinase in the activation of ternaryeom plexfactors[J].MolCellBiol，1997，17:2360-2371.
[69]RobinsonMJ，CobbMH.Mitogen-aetivated protein kinase path ways[J].Currop in CellB101，1997，9:180-186.
[70]Nemotos，ShengZ，LinA.opposin geffeets of junkina seand p38 mitogen-aetivated Proteinkin asesoncar diomyocyteshy PertroPhy[ Jl.MolCellBiol，1998，18:3518-3526.
[71]Ka1imanP，VinalsF，TestarX，etal.PhosPhatidy linositol3-kinaseinhibitors bloek differentiation of skeletal muscle cells[J].JBioChemi.1996，32(271):19146-19151.
[72]MenaseheP，HagegeAA， VilquinJT，etal.Autologouss keletal my oblasttrans-plantation for severe Postin farction left ventrieulardysfunetion[J」 .JAmCollCardiol，2003，41(7):1078-1083.
[73]DevineSM，HoffmanR.Role of mesenehymal stemeell sinhematopoietie stemeell trans plantation[Jl.CurropinHematol.2000，7(6):358-363.[74〕胡兵，安红梅，沈克，等.中医肾理论与干细胞研究[JJ.中国组织工程研究与临床康康复， 2008，12(3):598-601.
[75]郑景辉，李勇华，袁肇凯.骨髓间充质细胞中医药干预研究的现状及其前景和策略[J].辽宁中医药大学学报，2009，11(s):35-37.[76」汪朝晖，洗绍祥，杨忠奇，等.人参总皂贰诱导骨髓间充质干细胞分化为，二肌样细胞的实验研究[J〕.广州中医药大学学报，2006，23(2):100-103.
[77]王康锋，张洪斌，张立娟.中医肾精理论与神经干细胞关系探讨[J].新中医，2005，37(12):76-77.
[78]张进，徐志伟，陈群，等.干细胞与中医基础理论中的先天之精学说[J].中国组织工程研究临床康复，2006，10(7):189-192.
[79]FriedensteinAJ， PetrakovaKV， KurolesovaAl，etal.HeterotoPic of bonemarrow:Analy sis of preeursoreells forost eogenieand hema to Poietie tissues[J].TransPlantation，1968，6(2):230-247.
[80]马力，刘大军，李德天，等.不同分离方法及培养条件对兔骨髓间充质干细胞生长增殖及生物学特性的影响[J].中国组织工程研究与临床康复，2008，12(38):7401-7406.
[81]卢新政，张晓文，黄峻，等.大鼠骨髓间充质干细胞培养方法的改良[J].中国心血管杂志2004，9(1):48-51.
[82]YoshimuraH，MunetaT，NimuraA，eta1.Comparisonofratmesenehyma1stemcel1sderivedfrombonemarrow，synovium，Periosteum，adiPosetissueandmusele[J」.Ce11TissueRes，2007，327(3):449一462.
[83]HughesGC，PostMJ，SimonsM，etal.TranslationalPhysiology:Poreinemodelsofhumaneoronaryarterydisease:imPlicationsforPreelinicaltrialsoftherapeutieangigenesisfJ}.ApplPhysiol，2003，94:1689一1701.
[84]PaganiFD，SimonianHD，ZawadzkaA，etal.AutologousSkeletalMyoblastsTransPlantedtoisehemiadamagedmyocardiuminHumans〔J].JournaloftheAmericanCollegeofCardiology，2003，(41):879一888.
[85]黄国倩，刘虹，舒先红，等.超声心动图评价粒细胞集落刺激因子动员自身骨髓干细胞对心肌梗死后左心室重构及心功能的影响[J1.中华超声影像学杂志，2004，13(11):848-852.
[86]和岚，蒋文跃，毛腾敏.注射肾上腺素模拟”气滞”复制急、慢性血疥模型初探[J].中国中西医结合杂志，2004，24(3):244一246.
[87]ZhangH，XuC，LiangL，etal.Participation of bonemarrow derivedme senchymal stemcell sine or nealepi the lial wound healing[J].Invest.ophthalmol.Vis.Sei，2008，49:6071.
[88]李玉玲，唐俊明，潘国栋，等.DAPI标记的骨髓间充质干细胞体内外示踪实验研究[J].员卜阳医学院学报，2005，24(5):257-260.
[89]刘伯轩，刘师伟，王建华，等.DAPI标记的骨髓间充质干细胞体内外示踪实验研究[J]中国药物与临床，2009，9(8):703一705.
[90]orieD，KajsturaJ，ChimentiS，etal.Bonemarrow cell sregener ateinfraeted myoeardium[J].Natrue，2001，410:701-705.
[91]牛丽丽，祝善俊，张树林，等.同种异体骨髓间充质干细胞在大鼠心脏的迁移及分化特点[J].中华心血管病杂志，2004，32(增刊):186-189.
[92]GrossJ，LapiereCM.Collage nolyticae tivityi namphibian tissues:atissuecul tureassay[J].proeNatl AeadSciuSA.1962，15(48):1014-22.
[93]李建生，刘敬霞，孙捷，等.大黄普元对骨髓间充质干细胞移植脑缺血大鼠基质金属蛋白酶的影响[J].中西医结合学报，2008，6(s):510一816.
[94]ChengS，LovettDH.GelatinaseA(MMP一2)15necessaryandsufficientforrenaltubulareellepithelialmesenehymaltransformation[J].AmJPathol，2003，162(6):1937-1949.
[95]郑丽芳，张红，张小乔.GF一pl对骨髓间充质干细胞迁移力及Snail表达的影响[J].细胞生物学杂志，2005，30:222一228.
[96]SpinaleFG，CokerML，KrombaehSR，etal.Matrixmetal loproteinaseinhibition during the develoPment of eongestive heart failure:effectsonleftventrieulardimensionsandfunction[J].CircRes，1999，85:364一376.
[97]SpinaleFG，CokerML，HeungLJ，etal.Amatrixme talloproteinase induction/aetivation system exist sin the human left ventrieular myoear diumandlau Pregulate dinheartfailure[J].Circulation，2000，102:1944-1949.
[98]NagayaN，KangavaK，etal.Transplantationofmesenehymalstemeellsimprovescardicfunctioninaratmodelofdilatedcardiomyopathy[J].Circulation2005，112(8):1128-1135.
[99]雷警瑜，王卫淑，姚巍.间充质干细胞移植对，。肌基质金属蛋白酶一9及抑制因子-1表达的影响〔J].临床，“血管病杂志，2009，25(6):453一456
[100]Ka1imanP，VinalsF，TestarX，etal.PhosPhatidylinositol-3kinaseinhibitorsb1oekdifferentiationofskeletalmuscleeells[J」.JBioChemi，1996，32(271):19146-19151.
[101]KrasilnikovMA.PhosPhatidy linosito-3kinase dePendent Pathways:the roleineon trolofeell growth，survival，andmalignanttransformation[J」.Bioehemistry，2000，65(1):59-671.
[102]JaiswalRK，JaiswalN，BruderSP，etallAdul thumanme senehymal stemeell di-fferrentiationto the osteogenicoradiPogeniclineage15regulatedbym1toZgenZactivatedproteinkinase[J].JBiolChem，2000，275(13):9645一9652.
[103]王劲，郭朝华，罗成基，等.成肌细胞增殖n分化的信号转导研究[J]1生命的化学，2002，22(1):18-20.
[104]王劲，罗成基，郭朝华，等.p38及P工3K的激活在工GF21促进BMSCs成肌分化中的作用[J].第三军医大学学报，2003，25(13):1137-1139.
[105]Hami doucheZ，HaLE，VaudinP，etal.FHLZ media tesdexamethasone-indueed mesenehy maleell differentiation intoo steoblastsbyaetivatingwnt/p-catenin Signaling-dependent RunxZex pression[J].FASEBJ，2008，22:3813一3822.
[106]Boeelli-TyndallC，BracciL，SchaerenS，eta1.Human bone marrowme senehyma stem cells and chondrocytespromoteand/orsuPPresstheinvitroProliferationoflymphoeytesstimulatedbyinterleukinsZ，7and15[Jl.AnnRheumDis，2009，68:1352-1359.
[107]MatsushitaK，WuY，okamotoy，etal.local reninang iotensin expression regulate shumanme senehymal stemcell differentiation toad ipocytes[J].Hypertension，200648:1095-1102.
[108]Aurichl，MuellerLP，AurichH，etderivedfrom human mesene by mala1.Functional integration The Patocytes Stem cell sintomouselivers[J].Gut，2007，56:405-415.
[109]MartinezC，HoffnannTJ，MarinoR，etal Human bone marrowme senehymal stromaleell sex Press then euralgang liosideGDZ:ofBMSCs[J].Blood，2007，109:anovelSurfacemarkerfortheidentification4245-4248.
[110]HakunoD，FukudaK.Bonemarrow-derivedregeneratedeardiomyocytes(CMGcells)expressfunetionaladrenergicandmusearinicrecePtor[J].Cireulation2002，105(3):380-386.
[111]StrauerBE，BrehmM，ZeusT，etal.Repairofin fracted myocardiumby auto logousin tracoronary mononuelearbonemarrowcelltransplantationinhumans[J].C1rculation，2002，106(15):1913一1918.
[112]Tomitas，MiekleDA，WeiselRD，etal.Improvedheartfunetion with my ogenesis and angiogenesisafter auto logouspo reinemarrow stromaleelltran splan tation[J].JThoracCardiovaseSurg，2002，123:1132-1140.
[113]秦伯未.谦斋医学讲稿〔M〕.上海科学技术出版社，新1版，1978:2此.
[114]袁肇凯，周泽泉.养心通脉片治疗冠心病心绞痛的临床研究[J].中药新药与临床药理，1998，9(1):19一21.
[115]史学军，闰文浩.养心通脉方治疗冠心病51例疗效观察[J].中国医刊，1999，34(7):50-51.
[116]衰肇凯，曹金枝，黄献平，等.养心通脉液对心肌缺血再灌注损伤大鼠冠状微循环的影响[J」.中西医结合心脑血管病杂志，2004，2(12):715-717.
[117]王东生，袁肇凯，黄献平，等.养心通脉片对动脉粥样硬化大鼠“疾癖”病理细胞凋亡的影响〔J〕.中医杂志，2004，45(12):937-940.
[118]卢芳国，衰肇凯，黄献平，等.养心通脉片对胰岛素抵抗综合征外周血单核细胞C-myc和PDGF一AmRNA的影响〔J].中西医结合心脑血管病杂志，2003，1(8):442-444.
[119]莫莉，黄献平，袁肇凯，等.养心通脉有效部位方对鸡胚绒毛尿囊膜血管新生的影响[J〕中药新药与临床药理，2009，20(4):315一319.
[120]哀肇凯，莫莉，黄献平，等.养心通脉有效部位方对心肌缺血大鼠心血管内皮细胞血管生成的影响[J]一。血管康复医学杂志，2009，18(2):180一184.
[121]ColterDC，ClassR，DIGirolamoCM，eta1.Rapidexpansionofreeye1ingstemce11sinculturesofPlastie一adherent cells from human bone marrow[J].ProcNatlAcadSeiUSA，2000，97(7):3213-3218.
[122]郭艳，史大卓，陈可冀.中医药结合干细胞移植治疗心肌损伤性疾病的前景[J].新中医[J].2005，37(5):88-89.
[123]oswaldJ，BoxbergerS，JorgensenB，etal.Mesenchymals temcellseanbe different iatedinto and othelial cell sinvitro[Jl.StemCells，2004，22:377-384.
[124]ShengXG，SongH，FengJZ，etal.VEGF-exPressingbone marrowme senchymal stemeell strans Plantationim Provedheart function of myoeardialinfare trabbits[J]South China Journalo fear diology，2006，6(1):14-17.
[125]洗绍祥，杨忠奇，汪朝晖，等.黄茂甲普体外诱导骨髓间充质千细胞分化为心肌样细胞的实验研究[J]广州中医药大学学报，2007，24(1):37-40.
[126]宋清，张晓文，卢新政，等.参三七皂普Rgl预适应对5-氮胞普诱导大鼠骨髓间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复，2007，11(24):4693-4697.
[127]章亭，谭允育，潘彦舒.四物汤对红细胞免疫及骨髓干细胞增殖能加勺影响[J].北京中医药大学学报，2000，23(1):36-38.
[128]王春燕，曾和平，周亚东，等.补阳还五汤乙醇、氯仿提取物挥发性成分及调控鼠骨髓间充质干细胞活性研究[J]华南师范大学学报(自然科学版)2007，3:88-92.
[129]黄明明，崔晓瑞，邵雪喜.虫草类的免疫药理研究工.人工发酵的虫草菌的免疫抑制作用的观察[J].中华器官移植杂志，1984，5(2):58.
[130]曹丰，贾国良，牛丽丽，等.移植时机对骨髓间充质干细胞修复梗死心肌的影响[J].第四军医大学学报，2005，26(1):22-27.
[131]SuzukiK，SmolenskiRT，JayakumarJ，etal.Heat shoek treatment enhanees graft cell surviva linske letal my oblast transplantationtotheheart[J].Circulation，2000，102(19SupP13):111216-111221.
[132]GnecchiM，HeH，NoiseuxN，etal.EvideneesuPportingparaerinehypothesisforAkt-modifiedme senehymal stemeell-media tedear diaeProtection and funetional improvement[J].FASEBJ，2006，20(6):661-669.
[133]Jiangs，HaiderHKh，IdrisNM，etal.Supportiveinteraetionbetweeneellsurvival5ignaling and angioeom Petent faetor senhaneesdonor cell survival and Promotesang iomyogenesis for eardiae repair[J].CireRes.ZO06，99(7):776-784.
[134]GneeehiM，HeH，NoiseuxN，etal.Evideneesup Porting paraer inehypo thesis for Akt-modifiedmesenchymalstemcell-mediatedeardiac Proteetionand functional improvement [J].FASEBJ，2006，20(6):661-669
[135]TangYL，TangY，ZhangC，etal.Improved graft mesenehy malstem cell surviva linsehemi cheart with ahypoxia-regulated Hemeoxygenase-1veetor[J].JAmColl Cardiol，2005，46(7):1339-1350.
[136]JoJl，NagayaN，MiyaharaY，etal.Transplantation of geneti cell yengineered mesenehymalin faretion:Stem cell sim Provescardiaefunetionbenefitofanovelnonviralvector[J].inrats with myoear dialCationized DextranTISsueEng，2007，13(2):313-322.
[137]LIW，MaN，OngLL，etal.Bcl-2engine ered BMSC sinhibitedaPoPtosis and imProved heart function[J].StemCells.2007，25(8):2118-2127.
[138]BartunekJ，CroissantJD，WijnsW，etal.Pretreatfnen to fadult bone marrowme sene by malstem cell swithcar diomyogenie growthfactorsandrePairofthechronieallyinfaretedmyocardium[J].AmJPhysiolHeartCirePhysiol，2007，292(2):1095-1104.
[139]Voytik-HarbinSL.Three-dimensional extra cell ularmatrixsub strates fore ellculture[Jl.Methods Cell Biol，2001，63:561-81.
[140]PaulD，SamuelSM，MaulikN.Mesen chymal stemeell:present chall enges and pros Pective cellular cardiomyo PlastyaP Proaches for myocardia1regeneration[J].AntioxidRedoxSignal，2009，11(8):1841-1855.
[141]ZhangY，ChuY，ShenW，etal.Effectofs-azaey tidine induction durationond differentiation of human first-trimes terfetalmesenehymalstemeellstowardscardiomyocyte-likeeells[J].InteraetCardiovascThoracSurg，2009，9(6):943-946.
[142]HuangJL，YangSX.Safetyofumbiliealeordblood-derivedmesenehymalstemeells(BMSCs)followings-azaser ineinductionandinhibitionofhumancardiaemyoeyteapoptosisbyBMSCs[J].SaudiMedJ，2009，30(9):1144-1149.
[143]JYeNS，ZhangRL，ZhaoYF，etal.Effectofs-azaeytidineon the proteinex Pression of Porcine bone marrowme senehymal stem cell sinvitro[J].Genomies Proteomics Bin for maties，2006，4(1):18-25.
[144]Moscosol，CentenoA，LoPezE，etal.Differentiation”invitro”ofinortalizedPoreinemesenehymalstemeellsintocardiomyoeytestransplantation[J].Transplant ation Proeeedings，2005，37(1):481-482.Primaryandforcell
[145]RameshwarP，QiuH，VatnerSF.Stem cell sincardiae repair inaninflam atory meroen vironment[J』.Minerva Cardioangiol，2010，58(1):127-146.
[146]李艳芬，孙兰军，赵英强，等.复方丹参滴丸干预骨髓间充质千细胞移植后，。肌再生的实验研究[J].中国中医药现代远程教育，2009，8(9):81-83.
[147]南亚均，李庆雯，王宇春，等.BMSc联合淫羊蓉普预处理的EPCS移植对AMI大鼠心功能的影响[J].山西中医，2009，25(2):44-46.
[148]HayloekDN，NilssonSK.Stem cell regulation by the hematopoie tiestem cellniehe[J」.Cellcycle，2005，4(10):1353-1355.
[149]NilssonSK，SinunonsPJ.Transplan table stem cells:hometo speeificniches[J].CurrOPin Hematol，2004，11(2):102-106.
[150]WitzIP.Yin-Yangactivities andvieiouse yelesin the tumor mieroen vironment[J].CaneerRes，2008，68(1):9-14.
[151]HengBC，HaiderHK，SimEK，etal. Strate gies fordirecting the differentiation of stem cell sin to the cardiomyogeni clineagein vitro[Jl.CardiovaseRes，2004，62(1):34-42.
[152]张金红，耿朝晖，段江燕，等.MTT比色法在贴壁细胞药物敏感型测定中的应用[J].南开大学学报(自然科学版)，1996，29(4):109.
[153]MakinoS，Fukudak，Miyoshi5.etal.Cardiomyo eyteseanbegener ated from marrow strofnal cell sinvitro[Jl.J-ClinInvest，1999，103(5):697-705.
[154]YoonJ，MinBG，KimYH，etal.Differentiation，engraftment and funetion aleffects of pre-treatedme senehymalstem cell sinaratmyoeardialinfaretmodel[Jl.AetaCardiol，2005，60(3):277-284.
[155]ShimWS，JiangS，WongP，etal.Exvivodifferentiationofhumanadu1tbonemarrow5temcellsintoeardiomyoeyte-likeeells[J].BioehemBioPhysResCo朋un，2004，324(2):481-488.
[156]肖诗亮，郑日善.不同方法诱导的骨髓间充质干细胞移植修复大鼠梗死心肌〔J].华中科技大学学报(医学版)2007，36(3):339-342.
[157]常静，雷寒，陈建斌，等.大鼠心肌细胞诱导人骨髓问充质千细胞向，。肌样细胞分化的研究[J].重庆医科大学学报，2007，32(9):907-910.
[158]Ba11SQ，Shuttle worth AC，Kielty CM，etal.Direct cell contactin fluenees bonemarrowmese neymalstem cell fate[J].IntJBioehemCellBio，2004，36(4):714-727.
[159]RangappaS Ent wistle JWWeehslerA Setal.Cardiomyoeyte media tedeontaetprograms human mesenchymal stem cells to express eardiogenie phenotype[J].J Thorac Cardio vasc Surg，2003，126(1):124-132.
[160]XuM，WaniM，DaiYS，etal.Differentiation of bone marrow stromal cells into the eardiae Phenoty Perequires intercellular communieation with myocytes[J].Circulation，2004，110(17):2658-2665.
[161]YuanYC，henLF，ZhangSY，etal.Differentiation of mesenehy malstemcell sintoeardi of nyogeniceell sunder the induetion of myoeardia leellly sate[Jl.Chinese Journal of Cardiology，2005，33(2):170-173.
[162]汪朝晖，洗绍祥，杨忠奇，等.人参总皂戒诱导骨髓间充质干细胞分化为，。肌样细胞的实验研究[J].广州中医药大学学报，2006，23(2):100-103.
[163]李艳芬，孙兰军，赵英强，等.复方丹参滴丸干预骨髓间充质干细胞移植后心肌再生的实验研究[J].中国中医药现代远程教育，2009，9:81-83.
[164]陆长青，冉黎，张全波，等.丹参诱导骨髓间充质干细胞向神经元样细胞分化及相关基因的表达[J].中国组织工程研究与临床康复，2005，12(47):9363-9366.
[165]LueianoCA，AnastasiosPS，KarlHS，etal.Cardiae repair with intramyo cardialin jeetiono fall ogeneic mesenehymal stemeell saftermyoear dialin faretion[J].PNAS，2005，102(32):11474-11479.
[166]XuM，WaniM，DaiYS，etal.Differentiation of bonemarrow stromal cellsinto the cardiae phenoty Perequires in tereellular conununication with myoeytes[J].C1rculation，2004，110(17):2658-2665.
[167]曹寅，潘呈.三维细胞培养技术研究进展[J].安徽医药，2006，20(3):221-222.
[168]DoueetC，Ernoul，ZhangY，etal.PlateletlysatesPromotemesenehymalstemeellexPansion:asafetysubstitutefor2005，animalserumincell-basedtheraPyaPplications[J].JCellPhysiol，205(2):228-236.
[169]PittengerMF，MaekayAM，BeckSC，etal.Multi-lineage protential of adult humanmes enehymal stemcells[J].seienee，1999，284:143-147.
[1]Colter DC,Class R, DiGirolamo CM, et al.Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow[J].Proc Natl Acad Sci USA,2000,97(7):3213-3218.
[2]郭艳,史大卓,陈可冀.中医药结合干细胞移植治疗心肌损伤性疾病的前景[J].新中医,2005,37(5)：88-89.
[3]Oswald J,Boxberger S, Jorgensen B, et al.Mesenchymal stem cells can be differentiated into endothelial cells in vitro[J]. Stem Cells, 2004,22:377-384.
[4]赵春华.干细胞原理、技术与临床[M].北京化学工业出版社,2006：6290-6291.
[5]Jiang W,Ma A, Wang T, et al. Homing and differentiation of mesenchymal stem cells delivered intravenously to ischemic myocardium in vivo:a time-series study[J]. Pflugers Arch,2006,453(1):43-52.
[6]Penna C, Raimondo S, Ronchi G, et al. Early homing of adult mesenchymal stem cells in normal and infarcted isolated beating hearts [J]. J Cell Mol Med,2008,12 (2):507-521.
[7]Doucet C,Ernou I,Zhang Y,Llense JR,et al.Platelet lysates promote mesenchymal stem cell expansion:a safety substitute for animal serum in cell-based therapy applications[J]. J Cell Physiol,2005,205(2): 228-236.
[8]Pittenger MF,Mackay AM, Beck SC,et al.Multi-lineage protential of adult human mesenchymal stem cells[J]. science,1999,284:143-147.
[9]肖诗亮,郑日善.不同方法诱导的骨髓间充质干细胞移植修复大鼠梗死心肌[J].华中科技大学学报(医学版)2007,36(3)：339-342.
[10]常静,雷寒,陈建斌,等.大鼠心肌细胞诱导人骨髓问充质干细胞向心肌样细胞分化的研究[J].重庆医科大学学报,2007,32(9)：907-910.
[11]李进领,周崇臣,孙春霞.灯盏细辛对骨髓间充质干细胞经静脉移植后对缺血性脑血管病的影响[J].中华实用中西医杂志,2007,20(23)：2025-2027.
[12]曾意荣,樊粤光,刘红,等.补肾活血中药对大鼠骨髓间充质干细胞体外增殖的影响[J].中药新药与临床药理,2007,18(3)：93-96.
[13]宋清,张晓文,卢新政,等.参三七皂苷Rgl预适应对5-氮胞苷诱导大鼠骨髓 间充质干细胞向心肌细胞转化的干预[J].中国组织工程研究与临床康复,2007,11(24)：4693-4697．
[14]章亭,谭允育,潘彦舒.四物汤对红细胞免疫及骨髓干细胞增殖能力的影响[J].北京中医药大学学报,2000,23(1)：36-38.
[15]王春燕,曾和平,周亚东,等.补阳还五汤乙醇、氯仿提取物挥发性成分及调控鼠骨髓间充质干细胞活性研究[J].华南师范大学学报(自然科学版),2007,3：88-92.
[16]黄明明,崔晓瑞,邵雪喜.虫草类的免疫药理研究Ⅰ.人工发酵的虫草菌的免疫抑制作用的观察[J].中华器官移植杂志,1984,5(2)：58.
[17]商洪才,张伯礼.缺血性中风急性期治疗现状及中医药的前景和策略[J].天津中医药,2005,22(12)：523-525.
[18]赵春华.干细胞原理、技术与临床[M].北京化学工业出版社,2006：625-627.
[19]贾延,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[J].中华医学杂志,2002,82(19)：1337-1341.
[20]阎德文,李海燕,吴清平,等.GSB中药血清对rBMSCs定向诱导分化为成骨细胞的影响[J].中国骨质疏松杂志,2007,13(7)：480-485.
[21]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[J].广州中医药大学学报,2007,24(1)：37-40.