氯吡格雷对兔髂腹动脉球囊损伤后血管内皮功能及平滑肌细胞的影响

Effects of clopidogrel on rabbit vascular endothelial function and smooth muscle cell in balloon injured iliac artery

目的:观察氯吡格雷对兔髂腹动脉球囊损伤后血管内皮功能及平滑肌细胞的影响。方法:37只新西兰兔随机分为正常组(n=7)、模型组(n=10)、氯吡格雷组(n=10)和阿托伐他汀组(n=10)。后3组通过高胆固醇喂养加髂动脉内膜剥脱建立动脉硬化模型。用生物化学技术检测血管成形术前后血清内皮素、一氧化氮浓度;用光镜、扫描电镜、透射电镜观察髂动脉损伤术后8周血管病理形态学改变并测量动脉内膜、中膜厚度和面积;用原位杂交技术检测血小板源性生长因子(PDGFmRNA)的表达。结果:①与模型组相比,氯吡格雷组和阿托伐他汀组血清内皮素浓度均降低,一氧化氮浓度均升高,差异有统计学意义(P<0.05,P<0.01);②氯吡格雷组和阿托伐他汀组动脉内膜厚度与内膜/中膜厚度比显著减少(P<0.05),内膜面积与内膜/中膜面积比明显减少(P<0.05);③PDGFmRNA的表达明显降低(P<0.05);④透射电镜见模型组血管损伤段平滑肌细胞胞体肥大,核增大,胞浆内线粒体、粗面内质网增加,胞内吞噬大量脂滴,呈“合成型”改变。氯吡格雷组和阿托伐他汀组增生的平滑肌细胞胞体及核较小,胞浆内粗面内质网、线粒体较少,胞内吞噬的脂滴较少,接近“收缩型”改变。扫描电镜示模型组内皮细胞排列紊乱,细胞连接破坏,有大量血小板和蛋白颗粒粘附;氯吡格雷组和阿托伐他汀组内皮细胞排列趋于正常,细胞连接恢复,血小板粘附少。结论:氯吡格雷能保护血管内皮功能,减轻球囊损伤后PDGF表达,抑制平滑肌细胞增殖和血管内膜增生,预防动脉粥样硬化的形成和发展。

Objective：To investigate the effects of clopidogrel on the iliac arterial endothelial function and smooth muscle cell in rabbits following balloon injury in iliac artery. Methods： Thirty -seven New Zealand rabbits were divided randomly into four groups： （1）normal group（ n = 7）;（2）model group （n = 10）; （3）Clopidogrel group （ n = 10） ; （4）atorvastatin group（ n = 10）. The arteriosclerosis model was established by linking with 1% cholesterol, 5 %lard and 3% yolk mixed forage following 8 weeks hypedipid diet and iliac endothelium denudation in the last three group. The plasma endothelin （ET） and nitric oxide（NO） concentrations in all rabbits were detected at the first and the end. All rabbits were killed after 8 weeks and the iliac arteries were removed. With HE staining, transmission electron microscope and scanning electron microscope, the artery morphology alteration was observed. The thickness, area of intima vs media were measured. The expression of platelet-derived growth factor （PDGFmRNA） was observed by hybridization in site method. Results： （1）After 8 weeks, the concentration of plasma ET in model group was higher than that in the clopidogrel and atorvastatin group, and the concentration of serum NO decreased obviously （ P 〈 0.05, P 〈 0.01） ;（2）In comparison with model group,the thickness, area of intima and the ratio of intima to media in clopidogrel group notably decreased （ P 〈 0. 05） ; （3）The expression of PDGF was significantly decreased （ P 〈 0.05） ,whereas there was no difference between clopidogrel and atorvastatin group; （4）Compared with model group, there were small vessel smooth muscle cell bodies and nucleus, less organs and myofilament contents, and less lipid droplet in clopidogrel and atorvastatin group under the transmission electron microscope, as well there were less platelet adherence in clopidogrel and atorvastatin group under scanning electron microscope. Condusion： Clopidogrel can effectively lessen the artery intima thickness and area after iliac endothelium denudation, which attributes to its inhibiting the PDGF expression and vessel smooth muscle cell proliferation, lessening the neointimal thickness and protecting the endothelial function in balloon -injured iliac arterial in rabbis, and preventing the form and development of arteriosclerosis.