内皮祖细胞移植对盐敏感性高血压大鼠心肌重构的影响研究

摘要

目的：分析内皮祖细胞（EPCs）移植对盐敏感性高血压大鼠心肌重构的影响。方法2015年1月—2016年2月于陕西省医学实验动物中心实验室进行实验，选取 Dahl 盐敏感大鼠66只，按随机数字表法分为正盐组、高盐组及 EPCs 移植组，各22只。正盐组大鼠每日予以0.3%氯化钠饮食，高盐组及 EPCs 移植组大鼠每日予以8%氯化钠饮食，其中 EPCs 移植组大鼠尾静脉注入 CM-Dil 标记的 EPCs；3组大鼠均连续喂养6周。观察第3代 EPCs 形态，比较3组大鼠心肌重构指标〔前壁厚度、后壁厚度、左心室舒张末期内径（LVDd）、左心室收缩末期内径（ LVDs）、短轴缩短率（FS）、左心室射血分数（LVEF）〕、新生毛细血管数、血管内皮细胞生长因子（VEGF）和 CXC 趋化因子配体16（CXCL16）基因及蛋白表达情况，同时观察3组大鼠 CM-Dil 标记阳性细胞及心肌纤维化情况。结果第3代EPCs 生长迅速、纯度高、形态单一。正盐组、EPCs 移植组大鼠前壁厚度、FS、LVEF 高于高盐组，LVDs、LVDd 低于高盐组（P <0.05）；EPCs 移植组大鼠前壁厚度、FS、LVEF 高于正盐组，LVDs 低于正盐组（ P <0.05）。正盐组、EPCs 移植组大鼠新生毛细血管数多于高盐组（P <0.05）；EPCs 移植组大鼠新生毛细血管数多于正盐组（ P <0.05）。正盐组、EPCs 移植组大鼠 VEGF mRNA 相对表达量高于高盐组，CXCL16 mRNA 相对表达量低于高盐组（ P <0.05）；EPCs 移植组大鼠 VEGF mRNA 相对表达量高于正盐组，CXCL16 mRNA 相对表达量低于正盐组（ P <0.05）。正盐组、EPCs 移植组大鼠 VEGF 蛋白相对表达量高于高盐组，CXCL16蛋白相对表达量低于高盐组（P <0.05）；EPCs 移植组大鼠 VEGF 蛋白相对表达量高于正盐组，CXCL16蛋白相对表达量低于正盐组（P <0.05）。EPCs 组大鼠可见 CM-Dil 标记阳性细胞，正盐组和高盐组大鼠未见 CM-Dil 标记阳性细胞。正盐组大鼠未发生心肌纤维化，高盐组及 EPCs 移植组大鼠均出现不同程度的心肌纤维化；与高盐组比较，EPCs 移植组心肌纤维化程度较轻，巨噬细胞浸润较少。结论 EPCs移植可延缓盐敏感性高血压大鼠心肌重构，增加大鼠心肌毛细血管数并上调 VEGF 基因及蛋白的表达，下调 CXCL16基因及蛋白的表达，有利于减少心肌纤维化。%Objective To investigate the impact of endothelial progenitor cells(EPCs)transplantation on myocardial remodelling of salt - sensitive hypertensive rats. Methods From January 2015 to February 2016,this experiment was carried out in the laboratory of Shaanxi Medical Laboratory Animal Center. A total of 66 salt - sensitive Dahl rats were selected and divided into A group,B group and C group according to random number table,each of 22 rats. Rats of A group were fed with 0. 3%sodium chloride diet,rats of B group and C group were fed with 0. 9% sodium chloride diet,meanwhile rats of C group received caudal vein injection of CM-Dil marked EPCs;rats of the three groups were continuously fed for 6 weeks. Cellular forms of the third generation of EPCs were observed,index of myocardial remodelling( anterior wall thickness,posterior wall thickness, LVDd,LVDs,FS and LVEF),number of new blood capillary,gene expression and protein expression of VEGF and CXCL16 were compared among the three groups,CM-Dil marked cells and incidence of myocardial fibrosis were observed. Results The third generation of EPCs growed quickly with high - purity and single - form. Anterior wall thickness,FS and LVEF of A group and C group were statistically significantly higher than those of B group,while LVDs and LVDd of A group and C group were statistically significantly lower than those of B group( P < 0. 05);anterior wall thickness,FS and LVEF of C group were statistically significantly higher than those of A group,while LVDs of C group was statistically significantly lower than that of A gruop(P < 0. 05). Number of new blood capillary of A group and C group was statistically significantly more than that of B group,respectively(P < 0. 05),meanwhile number of new blood capillary of C group was statistically significantly more than that of A group(P < 0. 05). Relative expression quantity of VEGF mRNA of A group and C group was statistically significantly higher than that of B group,respectively,while relative expression quantity of CXCL16 mRNA of A group and C group was statistically significantly lower than that of B group,respectively(P < 0. 05);relative expression quantity of VEGF mRNA of C group was statistically significantly higher than that of A group,while relative expression quantity of CXCL16 mRNA of C group was statistically significantly lower than that of A group(P < 0. 05). Relative expression quantity of VEGF protein of A group and C group was statistically significantly higher than that of B group,respectively,while relative expression quantity of CXCL16 protein of A group and C group was statistically significantly lower than that of B group,respectively( P < 0. 05);relative expression quantity of VEGF protein of C group was statistically significantly higher than that of A group,while relative expression quantity of CXCL16 protein of C group was statistically significantly lower than that of A group( P < 0. 05). CM-Dil marked positive cells were detected in C group,but not in A group or B group. No rat of A group found myocardial fibrosis,while rats of B group and C group found varying degrees of myocardial fibrosis,and the degree of myocardial fibrosis of C group was statistically significantly milder than that of B group,with less macrophages infiltration. Conclusion EPCs transplantation can delay the myocardial remodelling of salt - sensitive hypertensive rats,increase the myocardial blood capillary,up - regulate the gene expression and protein expression of VEGF,down - regulate the gene expression and protein expression of CXCL16,is helpful to reduce the incidence of myocardial fibrosis.