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Enhanced external counterpulsation promotes growth cytokines-mediated myocardial angiogenesis in a porcine model of hypercholesterolemia
作者单位:LUO Jing-yun(Division of Cardiology the First Affiliated Hospital, Sun Yat-sen University,Guangzhou, Guangdong 510080, China;Division of Cardiology, Guangzhou Red Cross Hospital,Guangzhou, Guangdong 510220, China);WU Gui-fu,XIE Qiang,MA Hong,ZHENG Zhen-sheng,DU Zhi-min(Division of Cardiology the First Affiliated Hospital, Sun Yat-sen University,Guangzhou, Guangdong 510080, China;The Key Laboratory on Assisted Circulation, the Ministry of Health, Guangzhou, Guangdong 510080, China);XIONG Yan,CHEN Guo-wei,YANG Da-ya,HE Xiao-hong(Division of Cardiology the First Affiliated Hospital, Sun Yat-sen University,Guangzhou, Guangdong 510080, China);ZHANG Yan(The Key Laboratory on Assisted Circulation, the Ministry of Health, Guangzhou, Guangdong 510080, China);LIU Dong-hong(Department of Ultrasound the First Affiliated Hospital, Sun Yat-sen University,Guangzhou, Guangdong 510080, China);WANG Kui-jian(Department of Biomedical Engineering the First Affiliated Hospital, Sun Yat-sen University,Guangzhou, Guangdong 510080, China) 
基金项目:China National Key Research Project of Science,国家自然科学基金 
摘    要:Background Enhanced external counterpulsation (EECP) improves ischemia in patients with refractory angina pectoris but the mechanism remains unclear. To explore the mechanisms of EECP action, we detected progenitor cells presenting any of the following markers CD34^+, CD29^+, and CD106^+.
Methods Growth cytokines-mediated progenitor cell mobilization and associated angiogenesis potential were assessed in a porcine model of hypercholesterolemia. Twenty-four male domestic swines were randomly assigned to 4 groups: normal diet (control, n=6), hypercholesterolemic diet (CHOL, n=-6), hypercholesterolemic diet with administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) (rhG-CSF, n=6), and hypercholesterolemic diet with EECP treatment (EECP, n=6). EECP was applied 2 hours every other day for a total of 36 hours. Serum levels of vascular endothelial growth factor (VEGF) and granulocyte colony-stimulating factor (G-CSF), peripheral blood progenitor cell counts, level of regional angiogenesis, and expression of VEGF and stromal cell derived factor l a (SDF-1α) in porcine myocardium were assessed, respectively.
Results A porcine model of hypercholesterolemia-induced arteriosclerosis was successfully established. There was no significant difference in serum levels of VEGF among the four groups. The serum levels of G-CSF in the EECP group increased significantly at week 15 and week 18 ((38.3±5.6) pg/ml at week 15 vs (26.2±3.7) pg/ml at week 12, P 〈0.05, and (46.9±6.1) pg/ml at week 18 vs (26.2±3.7) pg/ml at week 12, P 〈0.01). The serum levels of G-CSF in group 3 increased also significantly after receiving rhG-CSF injection for five days ((150±13.9) pg/ml at week 18 vs (24.8±5.4) pg/ml at week 12, P 〈0.01). Compared to other groups and other time points, progenitor celt counts increased significantly after 2-hour EECP treatment (108±13 vs 26±6 per 10^5 leukocytes, P 〈0.01), but not at week 18. The progenitor cell counts also increased significantly after subcutaneous injection of rhG-CSF for five days compared to the week 12 (baseline) (180±21 vs 25±7 per 10s leukocytes, P 〈0.01). There was no significant difference among the four groups at other time points. Moreover, the expression of VEGF and SDF-1α and the level of regional angiogenesis in myocardium increased significantly in both EECP and rhG-CSF groups.
Conclusions The results demonstrated that EECP could facilitate angiogenesis in the myocardium of atherosclerotic swines by increasing endogenous G-CSF, inducing an enhanced mobilization of progenitor cells and augmenting myocardial expression of VEGF and SDF-1α.

关 键 词:增强型体外反搏  高胆固醇血症  心肌血管生成  生长因子介导  猪模型  粒细胞集落刺激因子  血管内皮生长因子  致动脉粥样硬化

Enhanced external counterpulsation promotes growth cytokines-mediated myocardial angiogenesis in a porcine model of hypercholesterolemia
Authors:LUO Jing-yun  WU Gui-fu  XIONG Yan  CHEN Guo-wei  XIE Qiang  YANG Da-ya  HE Xiao-hong  ZHANG Yan  LIU Dong-hong  WANG Kui-jian  MA Hong  ZHENG Zhen-sheng  DU Zhi-min
Institution:[1]Division of Cardioiogy , the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China [2]Department of Ultrasound, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China [3]Department of Biomedical Engineering , the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China [4]Division of Cardiology, Guangzhou Red Cross Hospital, Guangzhou, Guangdong 510220, China [5]The Key Laboratory on Assisted Circulationl the Ministry of Health, Guangzhou, Guangdong 510080, China
Abstract:Background Enhanced external counterpulsation (EECP) improves ischemia in patients with refractory angina pectoris,but the mechanism remains unclear. To explore the mechanisms of EECP action, we detected progenitor cells presenting any of the following markers CD34+, CD29+, and CD106+.Methods Growth cytokines-mediated progenitor cell mobilization and associated angiogenesis potential were assessed in a porcine model of hypercholesterolemia. Twenty-four male domestic swines were randomly assigned to 4 groups: normal diet (control, n=6), hypercholesterolemic diet (CHOL, n=6), hypercholesterolemic diet with administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) (rhG-CSF, n=6), and hypercholesterolemJc diet with EECP treatment (EECP, n=6). EECP was applied 2 hours every other day for a total of 36 hours. Serum levels of vascular endothelial growth factor (VEGF) and granulocyte colony-stimulating factor (G-CSF), peripheral blood progenitor cell counts, level of regional angiogenesis, and expression of VEGF and stromal cell derived factor 1α (SDF-1α) in porcine myocardium were assessed, respectively.Results A porcine model of hypercholesterolemia-induced arteriosclerosis was successfully established. There was no significant difference in serum levels of VEGF among the four groups. The serum levels of G-CSF in the EECP group increased significantly at week 15 and week 18 ((38.3±5.6) pg/ml at week 15 vs (26.2±3.7) pg/ml at week 12, P <0.05,and (46.9±6.1) pg/ml at week 18 vs (26.2±3.7) pg/ml at week 12, P <0.01). The serum levels of G-CSF in group 3 increased also significantly after receiving rhG-CSF injection for five days ((150±13.9) pg/ml at week 18 vs (24.8±5.4)pg/ml at week 12, P <0.01). Compared to other groups and other time points, progenitor cell counts increased significantly after 2-hour EECP treatment (108±13 vs 26±6 per 105 leukocytes, P <0.01), but not at week 18. The progenitor cell counts also increased significantly after subcutaneous injection of rhG-CSF for five days compared to the week 12 (baseline) (180+91 vs 25±7 per 105 leukocytes, P <0.01). There was no significant difference among the four groups at other time points. Moreover, the expression of VEGF and SDF-1α and the level of regional angiogenesis in myocardium increased significantly in both EECP and rhG-CSF groups.Conclusions The results demonstrated that EECP could facilitate angiogenesis in the myocardium of atherosclerotic swines by increasing endogenous G-CSF, inducing an enhanced mobilization of progenitor cells and augmenting myocardial expression of VEGF and SDF-1α.
Keywords:angiogenesis  atherosclerosis  progenitor cells  enhanced external counterpulsation  shear stress  swine
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