树突状细胞与冠状动脉严重狭窄的关系及辛伐他汀的干预作用

The Relationship Between Dendritic Cells and Severe Coronary Stenosis and the Effect of Simvastatin Intervention

目的探讨冠状动脉严重狭窄患者动脉粥样硬化与树突状细胞关系及辛伐他汀免疫干预作用的可能机制。方法根据选择性冠状动脉造影结果,20例冠状动脉正常,无明显动脉粥样硬化斑块者为阴性对照组;20例冠状动脉严重狭窄,未服用他汀类药物者为动脉粥样硬化组;20例冠状动脉严重狭窄,服用辛伐他汀40mg/d,连续30天以上者为辛伐他汀组。分别抽取冠状动脉血20mL,采用密度梯度离心法分离单个核细胞,行树突状细胞体外培养扩增,采用流式细胞术检测树突状细胞免疫表型、平均荧光强度、刺激T淋巴细胞增殖能力的刺激指数;动脉粥样硬化组分为B1、B2两个亚组,B2组在树突状细胞培养第5天时加入100μmol/L辛伐他汀,其他培养与检测方法相同。结果冠状动脉血分离单个核细胞后经体外培养与扩增,均成功培养出典型树突状细胞,各组树突状细胞形态无差异;与阴性对照组比较,B1组CD1a阳性细胞比例、CD1a与CD80、CD83、CD86双阳性细胞比例、平均荧光强度、收获细胞总数、树突状细胞数量和刺激指数均明显增高(P<0.05);与B1组比较,B2组收获细胞总数无差异,但CD1a阳性细胞比例、CD1a与CD80、CD83、CD86双阳性细胞比例、平均荧光强度、树突状细胞数量和刺激指数均显著降低(P<0.05);与B1组比较,辛伐他汀组CD1a阳性细胞比例、CD1a与CD80、CD83、CD86双阳性细胞比例、平均荧光强度、收获细胞总数、树突状细胞数量和刺激指数均显著降低(P<0.05)。结论冠状动脉严重狭窄的冠心病患者病变冠状动脉中树突状细胞数量与表达强度显著升高,成熟度与刺激T淋巴细胞增殖能力增强,树突状细胞在冠状动脉严重狭窄病变中起着重要作用;辛伐他汀可抑制树突状细胞的定向分化,减少树突状细胞的数量,降低表达强度,抑制树突状细胞的成熟,降低树突状细胞刺激T淋巴细胞增殖的能力。

Aim To investigate the relationship between dendritic cells and severe atherosclerosis（As）of coronary heart disease patients and explore the possible immune mechanisms of Simvastatin.Methods According to the results of selective coronary angiography,20 negative control（no obvious atherosclerosis,group A）,20 severe coronary heart disease patients with severe coronary stenosis without any statins（As group,group B）and 20 severe coronary heart disease patients with severe coronary stenosis with simvastatins 40 mg daily for at least 30 days（Simvastatin group,group C）.20 mL blood samples were extracted through the angiographic catheter.Density gradient centrifugation was used to separate mononuclear cells（MNC）.MNC were cultured in vitro and differentiated to DC in vitro.In B2 group,100 μmol/L Simvastatin were added on day 5.Flow cytometry was used to detect the immune characteristics of DC.Percent of CD1a^＋,CD1a^＋CD80^＋,CD1a^＋CD83^＋,CD1a^＋CD86^＋ cells and the mean fluorescent intensity（MFI）were measured and recorded.Results After in vitro proliferation,mononuclear cells from coronary artery could differentiate into dendritic cells and the morphology of DC did not differ between groups;Compared with negative controls,the percent of CD1a^＋,CD1a^＋CD80^＋,CD1a^＋CD83^＋,CD1a^＋CD86^＋ cells,MFI,MNC number,DC number and stimulation index（SI）in group B1 were significantly increased（P〈0.05）;Compared with group B1,the MNC number in group B2 was not significantly higher,but the percent of CD1a^＋,CD1a^＋CD80^＋,CD1a^＋CD83＋,CD1a^＋CD86^＋cells,MFI,DC num-ber and stimulation index（SI）were decreased significantly（P〈0.05）;Compared with group B1,the percent of CD1a^＋,CD1a^＋CD80^＋,CD1a^＋CD83^＋,CD1a^＋CD86^＋cells,MFI,MNC number,DC number and stimulation index（SI）in group C were significantly decreased（P〈0.05）.Conclusion In coronary heart diseases with severe stenosis,the number of dendritic cells and the expression of CD molecules increased significantly,the maturation and stimulation intensi-ty enhanced and DC played a key role in the severe stenosis of coronary atherosclerosis;Simvastatin could inhibit the differ-entiation of DC,decrease the number of DC,reduce the expression intensity,inhibit the maturation of DC and thus reduce the stimulation function of DC.