蛭龙活血通瘀胶囊对心肌梗死模型大鼠外周血CD34~+、CD133~+的影响
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摘要
目的:通过检测心肌梗死模型大鼠血清中CD34~+、CD133~+的含量,探索蛭龙活血通瘀胶囊对心肌梗死大鼠骨髓的内皮祖细胞(endothelial progenitor cells EPCs)动员的影响以及可能的机制。方法:健康SD大鼠48只,随机分为假手术组、卡托普利组、模型组、蛭龙活血通瘀胶囊高剂量组(简称蛭龙高剂量组)、蛭龙活血通瘀胶囊中剂量组(简称蛭龙中剂量组)、蛭龙活血通瘀胶囊低剂量组(简称蛭龙低剂量组),每组8只。结扎冠脉前降支复制急性心肌梗死大鼠模型。应用蛭龙活血通瘀胶囊高中低剂量、卡托普利灌胃3周。各组大鼠于术后第0天、第1天、第3天、第5天、第7天、第20天尾静脉采血法取静脉血0.5~1ml,流式细胞仪分别检测血清中CD34~+、CD133~+的含量。处死大鼠,心脏直接采血3~5ml,分离出单核细胞(monocyte MNCs)层,培养4天,观察内皮祖细胞的形态。结果:血清中CD34~+、CD133~+的含量:与假手术组比较,各观察点模型组、卡托普利组、蛭龙高剂量组、蛭龙中剂量组、蛭龙低剂量组血清中CD34~+、CD133~+的含量均升高,差异具有统计学意义(P<0.001);与模型组比较,各观察点卡托普利组、蛭龙高剂量组、蛭龙中剂量组、蛭龙低剂量组血清中CD34~+、CD133~+含量均升高,差异具有统计学意义(P<0.001);与卡托普利组比较,各观察点蛭龙高剂量组血清中CD34~+、CD133~+含量无明显变化,差异无统计学意义(P>0.05);与蛭龙中剂量组比较,各观察点卡托普利组、蛭龙高剂量组血清中CD34~+、CD133~+含量均升高,差异具有统计学意义(P<0.001);与蛭龙低剂量组比较,各观察点卡托普利组、蛭龙高剂量组、蛭龙中剂量组血清中CD34~+、CD133~+含量均升高,差异具有统计学意义(P<0.001)。培养四天后,EPCs细胞体积较前变大,部分呈椭圆形,数量变多,多数贴壁生长。结论:(1)蛭龙活血通瘀胶囊能够有效促进大模型大鼠心肌梗死后EPCs向外周血动员。(2)蛭龙活血通瘀胶囊可能是通过增加外周血CD34~+、CD133~+的含量,上调血管内皮生长因子(VEGF)的水平,促进骨髓EPCs动员的效应。
Objective: By detecting myocardial infarction rats serum CD_(34)~+,CD_(133)~+ content,exploring Zhilong Huoxue Capsule on myocardial infarction in rats bone marrow endothelial progenitor cell mobilization effects and possible mechanism. Methods: 48 Healthy SD rats were randomly divided into controlled group,captopril group,model group,Zhilong Huoxue Capsule high-dose group,Zhilong Huoxue Capsule middle-dose group,Zhilong Huoxue Capsule low-dose group,each group had eight rats. Coronary artery ligation of copying the rat model of acute myocardial infarction( AMI). Control line open thoracic surgery,broke the pericardium and exposed to the heart,not ligation of coronary artery; Application of high,medium and low dose of Zhilong Huoxue Tongyu Capsule,captopril gavage for 3 weeks. The controlled group,the model group were amount of saline lavage for three weeks. Groups of rats in postoperative 0 day,1 day,3 days,5 days,7 days,20 days caudal vein blood sampling method in venous blood 0. 5 ~ 1 ml,flow cytometry instrument in serum was detected in the content of CD_(34)~+,CD_(133)~+ ; Put to death in the rat,the heart blood directly for 3 ~ 5 ml,isolate mononuclear cells layer,cultivating four days,observing the endothelial progenitor cells. Results: In serum of CD_(34)~+、CD_(133)~+ content: compared with the controlled group,each observation point model group,captopril group,Zhilong high-dose group,Zhilong middle-dose group,Zhilong low-dose group serum content of CD_(34)~+,CD_(133)~+ were higher,the difference was statistically significant( P < 0. 001); Compared with model group,the observation point the captopril group,Zhilong high-dose group,Zhilong middle-dose group,Zhilong low-dose group CD_(34)~+、CD_(133)~+ were higher,statistically significant difference( P < 0. 001); In comparison with the captopril group there was no significant change in the CD_(34)~+,CD_(133)~+ content in the Zhilong high-dose group serum of the level,and the difference was not statistically significant( P>0. 05). In comparison with the Zhilong middle-dose group,the CD_(34)~+、CD_(133)~+ content of the captopril group and Zhilong high-dose group serum of content were elevated,and the difference was statistically significant( P<0. 001). In comparison with Zhilong low-dose group,the CD_(34)~+,CD_(133)~+ content of the captopril group and Zhilong high-dose group serum of content were elevated,and the difference was statistically significant( P<0. 001). After four days of culture,the EPCs cell volume was larger than before,and the oval was partially enlarged in number. Conclusion:(1)Zhilong Huoxue Tongyu Capsule can effectively promote the rat bone marrow mobilization EPCs from peripheral blood after myocardial infarction.(2)Zhilong Huoxue Tongyu Capsule,which increases the level of VEGF by increasing the amount of CD_(34)~+,CD_(133)~+ in the serum,promotes the effect of the bone marrow EPCs mobilization.
Objective: By detecting myocardial infarction rats serum CD_(34)~+,CD_(133)~+ content,exploring Zhilong Huoxue Capsule on myocardial infarction in rats bone marrow endothelial progenitor cell mobilization effects and possible mechanism. Methods: 48 Healthy SD rats were randomly divided into controlled group,captopril group,model group,Zhilong Huoxue Capsule high-dose group,Zhilong Huoxue Capsule middle-dose group,Zhilong Huoxue Capsule low-dose group,each group had eight rats. Coronary artery ligation of copying the rat model of acute myocardial infarction( AMI). Control line open thoracic surgery,broke the pericardium and exposed to the heart,not ligation of coronary artery; Application of high,medium and low dose of Zhilong Huoxue Tongyu Capsule,captopril gavage for 3 weeks. The controlled group,the model group were amount of saline lavage for three weeks. Groups of rats in postoperative 0 day,1 day,3 days,5 days,7 days,20 days caudal vein blood sampling method in venous blood 0. 5 ~ 1 ml,flow cytometry instrument in serum was detected in the content of CD_(34)~+,CD_(133)~+ ; Put to death in the rat,the heart blood directly for 3 ~ 5 ml,isolate mononuclear cells layer,cultivating four days,observing the endothelial progenitor cells. Results: In serum of CD_(34)~+、CD_(133)~+ content: compared with the controlled group,each observation point model group,captopril group,Zhilong high-dose group,Zhilong middle-dose group,Zhilong low-dose group serum content of CD_(34)~+,CD_(133)~+ were higher,the difference was statistically significant( P < 0. 001); Compared with model group,the observation point the captopril group,Zhilong high-dose group,Zhilong middle-dose group,Zhilong low-dose group CD_(34)~+、CD_(133)~+ were higher,statistically significant difference( P < 0. 001); In comparison with the captopril group there was no significant change in the CD_(34)~+,CD_(133)~+ content in the Zhilong high-dose group serum of the level,and the difference was not statistically significant( P>0. 05). In comparison with the Zhilong middle-dose group,the CD_(34)~+、CD_(133)~+ content of the captopril group and Zhilong high-dose group serum of content were elevated,and the difference was statistically significant( P<0. 001). In comparison with Zhilong low-dose group,the CD_(34)~+,CD_(133)~+ content of the captopril group and Zhilong high-dose group serum of content were elevated,and the difference was statistically significant( P<0. 001). After four days of culture,the EPCs cell volume was larger than before,and the oval was partially enlarged in number. Conclusion:(1)Zhilong Huoxue Tongyu Capsule can effectively promote the rat bone marrow mobilization EPCs from peripheral blood after myocardial infarction.(2)Zhilong Huoxue Tongyu Capsule,which increases the level of VEGF by increasing the amount of CD_(34)~+,CD_(133)~+ in the serum,promotes the effect of the bone marrow EPCs mobilization.
引文
[1]陈伟伟,高润霖,刘力生,等.《中国心血管病报告2016》概要[J].中国循环杂志,2017,32(06):521~530
[2]郭艳娟,胡硕强,俞晓薇.急性心肌梗死患者急性期外周血内皮祖细胞和血清中血管内皮生长因子水平的意义[J].中国分子心脏病学杂志,2014,(02):896~898
[3]Krishnamurthy,P.,et al.Interleukin-10deficiency impairs bone marrow-derived endothelial progenitor cell survival and function in ischemic myocardium.Circ Res,2011,109(11):1280~1289
[4]Peichev,M.,et al.Expression of VEGFR-2and AC133by circulating human CD34(+)cells identifies a population of functional endothelial precursors.Blood,2000,95(3):952~958
[5]梁煜,林代华,王清.气虚血瘀是冠心病的病机关键释义[J].中医药学刊,2003,(04):588~599
[6]文小敏,袁晓清,贺运河.益气活血方对急性心肌梗死气虚血瘀型治疗作用的临床研究[J].南方医科大学学报,2008,(01):149~150
[7]李小林,杨思进,白雪.蛭龙活血通瘀胶囊对心肌梗死大鼠内皮祖细胞动员[J].泸州医学院学报,2014,(04):401~403
[8]Lutgens,E.,et al.Chronic myocardial infarction in the mouse:cardiac structural and functional changes.Cardiovasc Res,1999,41(3):586~593
[9]Makino,H.,et al.Decreased levels of circulating CD34(+)cells are associated with coronary heart disease in Japanese patients with type2diabetes.J Diabetes Investig,2015,6(4):473~478
[10]Bozdag-Turan,I.,et al.Relation between the frequency of CD34(+)bone marrow derived circulating progenitor cells and the number of diseased coronary arteries in patients with myocardial ischemia and diabetes.Cardiovasc Diabetol,2011,10:107
[11]Platet,N.,et al.Influence of oxygen tension on CD133phenotype in human glioma cell cultures.Cancer Lett,2007,258(2):286~290
[12]Mansour,S.,et al.COMPARE-AMI trial:comparison of intracoronary injection of CD133+bone marrow stem cells to placebo in patients after acute myocardial infarction and left ventricular dysfunction:study rationale and design.J Cardiovasc Transl Res,2010,3(2):153~159
[13]梁小卫,田福利,梁春,等.通心络体外促进人外周血内皮祖细胞的增殖、迁移、黏附的研究[J].国际心血管病杂志,2009,36(03):184~187
[14]Singh,N.,et al.ACE2/Ang-(1~7)/Mas axis stimulates vascular repair-relevant functions of CD34+cells.Am J Physiol Heart Circ Physiol,2015,309(10):1697~1707
[2]郭艳娟,胡硕强,俞晓薇.急性心肌梗死患者急性期外周血内皮祖细胞和血清中血管内皮生长因子水平的意义[J].中国分子心脏病学杂志,2014,(02):896~898
[3]Krishnamurthy,P.,et al.Interleukin-10deficiency impairs bone marrow-derived endothelial progenitor cell survival and function in ischemic myocardium.Circ Res,2011,109(11):1280~1289
[4]Peichev,M.,et al.Expression of VEGFR-2and AC133by circulating human CD34(+)cells identifies a population of functional endothelial precursors.Blood,2000,95(3):952~958
[5]梁煜,林代华,王清.气虚血瘀是冠心病的病机关键释义[J].中医药学刊,2003,(04):588~599
[6]文小敏,袁晓清,贺运河.益气活血方对急性心肌梗死气虚血瘀型治疗作用的临床研究[J].南方医科大学学报,2008,(01):149~150
[7]李小林,杨思进,白雪.蛭龙活血通瘀胶囊对心肌梗死大鼠内皮祖细胞动员[J].泸州医学院学报,2014,(04):401~403
[8]Lutgens,E.,et al.Chronic myocardial infarction in the mouse:cardiac structural and functional changes.Cardiovasc Res,1999,41(3):586~593
[9]Makino,H.,et al.Decreased levels of circulating CD34(+)cells are associated with coronary heart disease in Japanese patients with type2diabetes.J Diabetes Investig,2015,6(4):473~478
[10]Bozdag-Turan,I.,et al.Relation between the frequency of CD34(+)bone marrow derived circulating progenitor cells and the number of diseased coronary arteries in patients with myocardial ischemia and diabetes.Cardiovasc Diabetol,2011,10:107
[11]Platet,N.,et al.Influence of oxygen tension on CD133phenotype in human glioma cell cultures.Cancer Lett,2007,258(2):286~290
[12]Mansour,S.,et al.COMPARE-AMI trial:comparison of intracoronary injection of CD133+bone marrow stem cells to placebo in patients after acute myocardial infarction and left ventricular dysfunction:study rationale and design.J Cardiovasc Transl Res,2010,3(2):153~159
[13]梁小卫,田福利,梁春,等.通心络体外促进人外周血内皮祖细胞的增殖、迁移、黏附的研究[J].国际心血管病杂志,2009,36(03):184~187
[14]Singh,N.,et al.ACE2/Ang-(1~7)/Mas axis stimulates vascular repair-relevant functions of CD34+cells.Am J Physiol Heart Circ Physiol,2015,309(10):1697~1707