平滑肌细胞gp130在腹主动脉瘤中的作用机制研究  

Research on the mechanism of smooth muscle cell gp130 in abdominal aortic aneurysm

作　　者：韩迎春 杨雨琪 李玉琳 杜杰 HAN Yingchun;YANG Yuqi;LI Yulin;DU Jie(Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029)

机构地区：[1]首都医科大学附属北京安贞医院,北京市心肺血管疾病研究所,北京100029

出　　处：《北京生物医学工程》2024年第6期592-597,640,共7页Beijing Biomedical Engineering

基　　金：国家自然科学基金(32200925、81930014);北京市自然科学基金(7232012)资助。

摘　　要：目的 探讨IL-6细胞因子家族的共受体糖蛋白130(glycoprotein 130,gp130)对血管平滑肌细胞表型和腹主动脉瘤的影响,挖掘腹主动脉瘤防治的潜在新靶点。方法 使用他莫昔芬[0.1 mg/(g·d)]对Sm22-ER-Cre^(+)gp130^(loxP/loxP)小鼠连续腹腔注射5 d的方法诱导平滑肌细胞gp130的敲除。选取8周的平滑肌细胞gp130特异性敲除(CKO)和对照小鼠各17只,两组小鼠均通过联合Pcsk9腺相关病毒尾静脉注射(1×10^(11) v.g.)、高脂饲喂和血管紧张素II灌注[1 000 ng/(min·kg),28 d]制备小鼠腹主动脉瘤模型。血管超声检测腹主动脉直径变化,腹主动脉最大外径超过造模前小鼠的50%则判定为腹主动脉瘤形成。EVG染色检测血管弹力板断裂情况。实时定量PCR、蛋白免疫印迹和免疫组化染色检测血管的收缩相关基因及炎症相关基因的表达。胶原收缩实验检测小鼠原代血管平滑肌细胞的收缩功能。结果 与对照组相比,CKO小鼠腹主动脉瘤发生率降低(P<0.05)、血管直径下降(P<0.05)、弹力板断裂情况改善。CKO小鼠主动脉收缩相关基因(α-Sma、Sm22等)表达上调、炎症相关基因(Ccl2、Il-6等)下调、炎症细胞浸润减少。与对照细胞相比,分离的CKO原代血管平滑肌细胞收缩功能增强(P<0.05)。结论 平滑肌细胞特异性敲除gp130通过维持平滑肌细胞收缩表型和收缩功能、减轻血管炎症,在腹主动脉瘤进展中发挥保护作用。Objective To explore the impact of the interleukin-6 cytokine family co-receptor glycoprotein 130(gp130) on vascular smooth muscle cell phenotype and abdominal aortic aneurysm(AAA),identifying potential new targets for AAA prevention and treatment.Methods Smooth muscle cell gp130 knockout was induced by administering tamoxifen [0.1 mg/(g·d)] via intraperitoneal injection to Sm22-ER-Cre+gp130~(loxP/loxP) mice for five consecutive days.A total of 17 eight-week-old smooth muscle-specific gp130 knockout(CKO) and control mice were selected.Both groups underwent combined Pcsk9 adeno-associated virus via tail vein injection(1×10~(11) v.g.),high-fat feeding,and angiotensin II infusion [1 000 ng/(min·kg) for 28 days] to establish a mouse model of AAA.Vascular ultrasound was used to monitor changes in abdominal aortic diameter,with aneurysm formation defined as an increase of more than 50% in maximum outer diameter compared to baseline.EVG staining assessed the rupture of vascular elastic laminae.Real-time quantitative PCR,Western blotting,and immunohistochemistry analyzed the expression of contraction-related and inflammation-related genes.Collagen contraction assays evaluated the contraction function of primary mouse vascular smooth muscle cells.Results Compared to the control group,CKO mice showed a reduced incidence of AAA(P<0.05),decreased vascular diameter(P<0.05),and improved elastic lamina integrity.Expression of contraction-related genes(a-Sma,Sm22,etc.) was upregulated in CKO mice,while inflammation-related genes(Ccl2,Il-6,etc.) were downregulated,accompanied by reduced inflammatory cell infiltration.Isolated primary CKO vascular smooth muscle cells exhibited enhanced contraction function compared to control cells(P<0.05).Conclusions Smooth muscle cell-specific knockout of gp130 protects against the progression of AAA by maintaining smooth muscle cell contraction phenotype and function,as well as alleviating vascular inflammation.

关 键 词：腹主动脉瘤 平滑肌表型 收缩功能 血管炎症 GP130 

分 类 号：R318[医药卫生—生物医学工程]