AMD3100促进载脂蛋白E^(-/-)小鼠动脉粥样硬化斑块形成

AMD3100 Aggravates Apolipoprotein E^(-/-) Mice Atherosclerosis Plaque Formation

目的探讨AMD3100对载脂蛋白E-/-小鼠主动脉动脉粥样硬化斑块形成的影响及其可能机制。方法12只8周龄雄性载脂蛋白E-/-小鼠随机分为AMD3100组(2.5mg/kg,隔天注射)和对照组(PBS 0.1mL,隔天注射),高脂高胆固醇饲料喂养12周后,获取组织标本和骨髓细胞,石蜡切片HE染色检测小鼠主动脉根部动脉粥样硬化病变;通过计数典型内皮祖细胞克隆形成单位和观察次级集落单位的大小与细胞密度检测内皮祖细胞的克隆形成能力;逆转录聚合酶链反应和Western blotting检测内皮祖细胞CXCR4 mRNA和蛋白的表达。结果AMD3100组小鼠主动脉窦横切面粥样斑块面积与管腔面积之比与对照组比较增加了38.8%(37.2%±3.6%比26.8%±2.5%,P<0.05);AMD3100组小鼠骨髓源内皮祖细胞的克隆形成能力显著低于对照组(初级内皮祖细胞集落单位个数为9.67±2.16比21.83±2.64,次级内皮祖细胞集落单位个数为1.67±0.31比4.11±0.65,P<0.01);AMD3100组CXCR4 mRNA和蛋白的表达均显著低于对照组(P<0.01)。结论AMD3100促进载脂蛋白E-/-小鼠动脉粥样硬化斑块形成,可能与抑制骨髓源内皮祖细胞的克隆形成并下调CXCR4表达相关。

Aim To study the effects of AMD3100 on atherosclerosis and the possible mechanism in apolipoprotein E^-/- mice, Methods 12 male apolipoprotein E^-/- mice, 8 weeks old, were randomly divided into two groups： AMD3100 group （2.5 mg/kg, the next day intraperitoneal injection） and control group （PBS 0.1 mL, the next day intraperitoneal injection）, After fed with high fat and cholesterol western food for 12 weeks, all mice tissue specimen and bone marrow cells were harvested. The Hematoxylin/Eosin staining of paraffin section was performed to detect atherosclerotic plaque of aortic root. By counting the typical endothelial progenitor cells-colony forming units （EPC-CFU） and observing the size and cell density of secondary EPC-CFU, the clonality of endothelial progenitor cells was measured. The expression of CXCR4 mRNA and protein were examined by RT-PCR and Western blotting. Results Compared with control group, the area percentage of atherosclerotic lesion and vessel lumina increased 38.8% in AMD3100 treated apolipoprotein E^-/- mice （37.2% ± 3.6% vs 26.8% ± 2.5%, P 〈 0.05）. The clonality of endothelial progenitor cells derived from AMD3100 group decreased in comparison to control group （primary EPC-CFU： 9.67±2.16vs21.83±2.64, secondary EPC-CFUs： 1.67±0.31 vs4.11±0.65;P〈0.01）. The expression of CXCR4 mRNA and protein of AMD3100 group were also reduced （ P 〈 0.01 ）. Conclusions The atherosclerotic lesion was aggravated by administration of AMD3100 in apolipoprotein E^-/- mice. Possible mechanism of this action for AMD3100 are associated with the inhibition of the clonality of bone marrow endothelial progenitor cells and down-regulatipon of CXCR4 expression on endothelial progenitor cells.