血管生成素样蛋白3基因敲低减轻胸主动脉瘤的发生和发展

Gene knockdown of angiopoietin-like protein 3 reduces the occurrence and development of thoracic aortic aneurysms

目的探讨血管生成素样蛋白3(ANGPTL3)在胸主动脉瘤(TAA)发生和发展中的作用机制。方法选取于北京安贞医院进行开放性手术修复的TAA患者为实验组(n=6),对照组主动脉组织来自心脏移植供体(n=6)。另取3周龄雄性C57BL/6J小鼠30只,随机分为对照(control)和β-氨基丙腈酯(BAPN)诱导的TAA模型组(每组n=15),4周后取小鼠主动脉组织。免疫组化检测人胸主动脉瘤组织中ANGPTL3的表达;免疫组化和Western blot检测小鼠主动脉组织中ANGPTL3、IL-6、MMP2、MMP9表达;TUNEL检测凋亡;免疫荧光共定位检测ANGPTL3在血管组织中的定位;用ANGPTL3 siRNA转染人主动脉平滑肌细胞;观察ANGPTL3抑制后对AngⅡ诱导平滑肌细胞凋亡及炎性反应的作用。结果在胸主动脉瘤患者和TAA小鼠的主动脉组织内,ANGPTL3蛋白表达水平显著增加(P<0.05);血管平滑肌细胞发生凋亡,同时炎性因子IL-1β、MMP2、MMP9蛋白表达均显著增加(P<0.05);ANGPTL3敲低可显著减轻AngⅡ诱导的平滑肌细胞凋亡和炎性因子分泌(P<0.05)。结论ANGPTL3在胸主动脉瘤发生发展过程中蛋白表达水平增高,而降低ANGPTL3蛋白表达水平可减轻平滑肌细胞凋亡和炎性反应。提示靶向ANGPTL3可抑制胸主动脉瘤的发生和发展。

Objective To explore the role of angiopoietin-like protein 3(ANGPTL3)in the development of thoracic aortic aneurysm(TAA)and its possible mechanism.Methods The TAA patients who underwent open surgical repair in Beijing Anzhen Hospital were selected as the experimental group(n=6),and the aortic tissue in the control group was obtained from a heart transplant donor(n=6).Another 303-week-old male C57BL/6J mice were randomly divided into control(control)andβ-aminopropionitrile(BAPN)-induced TAA model groups(n=15 per group).The aortic tissue was taken 4 weeks later.Immunohistochemical method was used to detect the expression of ANGPTL3 in human thoracic aortic aneurysm tissue,immunohistochemistry were used to detect the expression ofANGPTL3,IL-6,MMP2,and MMP9 in mouse aortic tissue,and TUNEL method was used to detect apoptosis.Immunofluorescence co-localization was used to detect the localization of ANGPTL3 in vascular tissues.Transfect human aortic smooth muscle cells with ANGPTL3 siRNA.Observe the effect of ANGPTL3 knockdown on AngⅡ-induced smooth muscle cell apoptosis and inflammatory response.Results In the aortic tissues of patients with thoracic aortic aneurysm and TAA mice,the expression of ANGPTL3 protein increased significantly(P<0.05),vascular smooth muscle cell apoptosis,and the protein expressions of inflammatory factors IL-1β,MMP2,and MMP9 were all significantly increased(P<0.05).In addition,ANGPTL3 knockdown can significantly reduce AngⅡ-induced smooth muscle cell apoptosis and inflammatory factor secretion(P<0.05).Conclusions The ANGPTL3 protein level is increased in the development of thoracic aortic aneurysm.Inhibition of ANGPTL3 protein expression can reduce the cell apoptosis and inflammation of smooth muscle cells.It suggests that targeting ANGPTL3 may inhibit the development of thoracic aortic aneurysms by inhibiting cell apoptosis and inflammation in smooth muscle.