Zhao Gui-Feng, Fan Ying-Chang, Jiang Xi-Juan
Department of Cardiology, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 May;32(5):671-5.
To observe the proliferation state of transplanted cells in acute myocardial infarction (AMI) rats, and the endothelial progenitor cells (EPCs) preconditioned by salvianolic acid B in different ratios with the bone mesenchymal stem cells (BMSCs).
The cultivation and purification of EPCs were performed by density-gradient centrifugation and plastic adherence method. Two types of cells were identified by immunocytochemical method (CD34, CD133, and CD44). The rat model of AMI was prepared by ligation of left anterior descending artery. The EPCs were pre-treated with the optimal concentration of salvianolic acid B (8 microg/ mL). They were mixed with BMSCs in different proportions (EPCs/BMSCs in the ratio of 1:1, 2:1, 4:1, and 8:1, respectively). BMSCs and EPCs were injected into the myocardial infarction area. The infarcted area was determined by the N-BT staining and hematoxylin-eosin staining. The expression of Ki-67 was detected by immunohistochemical assay.
Compared with the model group (19.60% +/- 3.23%), the myocardial infarction area of each implanted group obviously decreased (P < 0.05). Of them, the decrease was most obvious in the 4:1 group (11.37% +/- 2.18%) and the 8:1 group (9.23% +/- 2.35%, P < 0.05). Compared with the model group (cell/high magnification, 5.17 +/- 2.31), the Ki-67 positive cell number of each implanted groups significantly increased (P < 0.05). Of them, the Ki-67 positive cell number was obviously higher in the 8:1 group (15.00 +/- 3.16, P < 0.05).
EPCs pretreated by salvianolic acid B combined with BMSCs could reduce the myocardial infarcted area, improve the proliferation of BMSCs in the peripheral infarction and local ischemia. Besides, along with the increase of the implant proportion of EPCs, the infarct area was gradually reduced, and the proliferative expression was gradually enhanced.
观察急性心肌梗死（AMI）大鼠移植细胞的增殖状态，以及经丹酚酸B预处理的内皮祖细胞（EPCs）与骨髓间充质干细胞（BMSCs）不同比例混合后的情况。
采用密度梯度离心法和塑料贴壁法进行EPCs的培养与纯化。通过免疫细胞化学方法（CD34、CD133和CD44）鉴定两种细胞。采用结扎左冠状动脉前降支的方法制备大鼠AMI模型。用最佳浓度的丹酚酸B（8μg/mL）对EPCs进行预处理。将其与BMSCs按不同比例混合（EPCs/BMSCs比例分别为1:1、2:1、4:1和8:1）。将BMSCs和EPCs注入心肌梗死区域。通过N-BT染色和苏木精-伊红染色测定梗死面积。采用免疫组织化学分析法检测Ki-67的表达。
与模型组（19.60%±3.23%）相比，各植入组的心肌梗死面积均明显减小（P<0.05）。其中，4:1组（11.37%±2.18%）和8:1组（9.23%±2.35%，P<0.05）减小最为明显。与模型组（细胞/高倍视野，5.17±2.31）相比，各植入组的Ki-67阳性细胞数均显著增加（P<0.05）。其中，8:1组的Ki-67阳性细胞数明显更高（15.00±3.16，P<0.05）。
经丹酚酸B预处理的EPCs联合BMSCs可减小心肌梗死面积，促进BMSCs在梗死周边和局部缺血区域的增殖。此外，随着EPCs植入比例的增加，梗死面积逐渐减小，增殖表达逐渐增强。
