树突状细胞与冠状动脉侧支循环关系的研究

目的：探讨冠状动脉严重狭窄冠心病患者冠状动脉血树突状细胞（dendritic cells, DC）的数量、表型、功能状态与冠状动脉侧支循环的关系。方法：收集40例冠状动脉严重狭窄冠心病患者,根据冠状动脉侧支循环（coronary collateral circulation, CCC）形成情况分为CCC形成组（A组,n=22）和无CCC形成组（B组,n=18）。采用密度梯度离心法分离冠状动脉血单个核细胞,行DC体外培养与扩增,观察DC形态,检测收获细胞总数和DC数量;采用流式细胞仪检测DC表型与平均荧光强度（mean fluorescence intensity,MFI）,采用同种异体混合淋巴细胞反应检测DC刺激T淋巴细胞增殖能力,计算刺激指数(stimulation index,SI)。结果：（1）冠状动脉血分离单个核细胞后成功培养出典型DC,两组DC形态上无差异;（2）A组收获细胞数为 （3.95±1.41）×106,B组收获细胞数为（2.76±0.92）×106,A组收获细胞数显著增多,差异有统计学意义（P=0.003）;（3）A组DC数为（1.54±0.96）×106,B组DC数为（0.99±0.46）×106,A组DC数明显增多,差异有统计学意义（P=0.033）;（4）两组CD1a阳性细胞比例; MFI, CD1a与CD80, CD83, CD86双阳性细胞比例及MFI差异均无统计学意义（P>0.05）;（5）A组SI为4.96±2.30,B组SI为2.66±1.04,A组SI显著增高,差异有统计学意义（P=0.0003）。结论：在冠状动脉严重狭窄的冠心病患者中,冠状动脉侧支形成患者DC的数量显著增加,刺激T淋巴细胞增殖的能力增强。

Dendritic cells and coronary collateral circulation in coronary heart disease

ObjectiveTo determine the relationship between the number,phenotype and functional status of dendritic cells (DCs) and coronary collateral circulation (CCC) in coronary heart disease (CHD). MethodsForty patients with severe coronary stenosis were recruited and divided into a CCC formation group (Group A, n=22) and a non CCC formation group (Group B, n=18). Density gradient centrifugation was applied to separate the mononuclear cells (MNCs) from coronary artery blood samples, and MNCs were cultured and proliferated in vitro.The morphology of DCs was observed under converted microscope. The number of harvested cells and DCs was counted by hematocytometer. Flow cytometry was applied to investigate the phenotype and the mean fluorescence intensity (MFI). Mixed lymphocyte reaction was used to test the function of DCs to stimulate the proliferation of T lymphocytes. Stimulation index (SI) was calculated and compared. Results(1) After in vitro proliferation, DCs were cultured successfully from the mononuclear cells from coronary artery blood samples and the morphology of DCs was not different in the 2 groups. (2) The number of mononuclear cells(MNC no) was (3.95±1.41)×106, in the CCC group and (2.76±0.92)×106 in the non CCC group. The MNC number was significantly increased in the CCC group （P=0.003）. (3) The number of DCs was (1.54±0.96)×106 in the CCC group, and (0.99±0.46)×106 in the non CCC group（P=0.033）. (4)There was no statistical significance in the percent of CD1a+, CD1a+CD80+, CD1a+CD83+, CD1a+CD86+ cells, and MFI in the 2 groups（P>0.05）. (5) SI was 4.96±2.30 in the CCC group, whereas 2.66±1.04 in the non CCC group. The SI in the CCC group increased significantly（P=0.0003）. ConclusionIn CHD patients with severe coronary stenosis, patients with CCC formation have higher number of DCs and stronger potential of T lymphocyte stimulation.