目的:观察冠状动脉(冠脉)外周脂肪及冠脉外膜对动脉粥样硬化的影响。方法:血管外膜分组:Ⅰ组:正常对照冠脉(6例);Ⅱ组:冠心病者冠脉(5例);Ⅲ组:冠心病者乳内动脉(12例);Ⅳ组:冠心病者桡动脉(6例);Ⅴ组:冠心病者大隐静脉(14例)。冠脉外周脂肪分组:冠心病组(16例),对照组(6例)。各组分别行血管的苏木精-伊红染色及CD68^+抗体标记的免疫组化检测;冠脉外周脂肪脂联素及TNF-α的mRNA水平检测及CD68^+抗体免疫组化检测。脂多糖(100ug/L)及硬脂酸(0.5mmol/L)刺激培养的对照组冠脉外周脂肪细胞,检测上清液TNF—α及IL-6浓度。结果:与其他血管外膜比较,Ⅱ组冠脉外膜可见明显的巨噬细胞聚集带。冠脉外周脂肪内浸润巨噬细胞数,冠心病组:(39±7.1)个/400倍,对照组:(12±4.3)个/400倍,前者巨噬细胞浸润明显较后者密集,P〈0.05;冠心病组脂联素mRNA表达明显降低,而TNF-α的表达明显增高。对照组冠脉外周脂肪细胞经硬脂酸及脂多糖刺激后,上清液TNF-α及IL-6浓度明显增高(P〈0.05)。结论:冠心病者冠脉外周脂肪及冠脉外膜炎性细胞局部浸润产生的炎症反应促进了冠脉病变的形成。
Objective:Perivascular adipose tissue secrete a number of adipocytokines. The aim of the study was to assess adipocytokine-vascular ineractions and associations between perivascular adipose tissue and coronary artery atherosclerosis. Method:Fourteen coronary artery disease (CAD) patients undergoing open heart surgery and eleven donors were chosen, vasculars in different segments were collected and divided into Ⅰ , Ⅱ,Ⅲ , Ⅳ ,and Ⅴ group for hematoxylin-eosin staining and immunohistological analysis. Perivascular adipose tissue were taken near the proximal tract of the right coronary artery before cardiopulmonary bypass, detected by using RT-PCR and immunohistocbemistry. Perivascular adipose cells were cultured in M199 medium for 24 h, culture medium was changed and the adipocytes were grown for 6 d. Stearic acid (SA) of the final concentration of 0.5 mmol/L and LPS 100 ug/L (the final concentration) respectively were added, 24 h later the concentration of TNF-α and IL-6 in the supernatant were detected. Result: Macrophages (CD68^+ cells) accumulated preferentially at the interface between the perivascular adipose tissue and adventitia of atherosclerotic aortas. The quantity of CD68^+ cell in perivascular adipose tissue of CAD group was significantly higher than non-CAD group. Adiponectin mRNA expression was 3 fold higher in perivascular adipose tissue of non-CAD group than CAD group (P〈0.05). Furthermore, the level of TNF-α mRNA in the perivascular adipose tissue of non-CAD group was also significantly lower than CAD group (P〈0.05). SA and LPS stimulation increased the concentration of TNF-α and IL-6 in the supernatant. Conclusion: Inflammation that occurs locally in perivaseular adipose tissue and adventitia of coronary artery contributed to the pathogenesis of CAD.