作者：姚弈伟，刘亚峰，陈淦一，王晓棣，陈鑫

单位：

210006江苏省南京市，南京医科大学附属南京医院 南京市第一医院心胸外科 通信作者：陈鑫，E-mail：stevecx@njmu.edu.cn

Units：

Department of Cardiothoracic Surgery, Nanjing First Hospital, Nanjing Medical University/Nanjing First Hospital, Nanjing210006, China Corresponding author: CHEN Xin, E-mail: stevecx@njmu.edu.cn

关键词：

主动脉瘤,腹; 细胞焦亡; 血管紧张素Ⅱ; 肌细胞,平滑肌; NLRP3; GSDMD; Caspase-1; 白介素18;

Keywords：

Aortic aneurysm, abdominal; Pyroptosis; Angiotensin Ⅱ; Myocytes, smooth muscle; NLRP3; GSDMD;Caspase-1; Interieukin-18

CLC：

DOI：

10.12114/j.issn.1008-5971.2022.00.111

Funds：

国家自然科学基金青年科学基金项目（81900417）

摘要：

目的 分析细胞焦亡在血管紧张素Ⅱ诱导的腹主动脉瘤（AAA）发生和发展中的作用。方法 2019年9月至2021年9月，将50只C57BL/6雄性野生型小鼠按照抽签法随机分为对照组（n=24）和血管紧张素Ⅱ组（n=26），所有小鼠在肩胛骨中部皮下植入渗透微型泵注入0.9%氯化钠溶液（对照组）或血管紧张素Ⅱ（血管紧张素Ⅱ组），以1 mg·kg-1·min-1的剂量持续灌注28 d。肉眼、腹主动脉超声检查、HE染色、马松三色染色及VVG染色、免疫荧光染色观察两组腹主动脉大体形态、动脉直径、管腔及动脉壁、弹力纤维破坏、胶原纤维沉积情况及细胞焦亡相关蛋白（NLRP3、GSDMD、Caspase-1和IL-18）表达情况。小鼠主动脉血管平滑肌细胞（MOVAS）分别加入DMEM（对照组）或血管紧张素Ⅱ（1μmol/L）(血管紧张素Ⅱ组)培养，采用Western blotting法检测MOVAS中细胞焦亡相关蛋白表达水平。结果 灌注28 d后，对照组小鼠均无动脉瘤形成，血管紧张素Ⅱ组小鼠发生动脉瘤。腹主动脉超声检查显示，血管紧张素Ⅱ组小鼠腹主动脉较对照组明显扩张。HE染色显示，血管紧张素Ⅱ组小鼠的腹主动脉管腔较对照组扩大，动脉壁变薄。马松三色染色和VVG染色显示，血管紧张素Ⅱ组小鼠弹力纤维降解，与AAA形成相关的胶原沉积明显增多。免疫荧光染色显示，血管紧张素Ⅱ组小鼠腹主动脉组织NLRP3、GSDMD、Caspase-1、IL-18蛋白表达增加。Western blotting法检测结果显示，对照组和血管紧张素Ⅱ组MOVAS中IL-18蛋白表达水平比较，差异无统计学意义（P>0.05）；血管紧张素Ⅱ组MOVAS中NLRP3、GSDMD、Caspase-1蛋白表达水平高于对照组（P<0.05）。结论 血管紧张素Ⅱ诱导的AAA形成与细胞焦亡相关，其主要是通过减少血管中膜层平滑肌细胞数量和促进炎症微环境导致血管壁弹性层被破坏，细胞焦亡在AAA的发生和发展过程中发挥重要作用。

Abstract：

【Abstract】　Objective To analyze the effect of pyroptosis in the formation and progression of angiotensin Ⅱ-induced abdominal aortic aneurysm (AAA) . Methods From September 2019 to September 2021, 50 C57BL/6 male wild-typemice were randomly divided into the control group (n=24) and the angiotensin Ⅱ group (n=26) by drawing lots. All mice weresubcutaneously implanted with osmotic micropumps in the middle of scapula and injected with 0.9% sodium chloride solution(control group) or angiotensin Ⅱ (angiotensin Ⅱ group) at a dose of 1 mg·kg-1·min-1 for 28 d. Macroscopic observation,abdominal aorta ultrasonography, HE staining, Masson staining, VVG staining and immunofluorescence staining were used toanalyze the abdominal general morphology, artery diameter, lumen and arterial wall, elastic fiber destruction, collagen deposition,and pyroptosis-related protein expression (NLRP3, GSDMD, Caspase-1 and IL-18) in the two groups. Mouse aortic smoothmuscle cells (MOVAS) was added with DMEM (control group) or angiotensin Ⅱ (1 μmol/L) (angiotensin Ⅱ group) , and thelevels of pyroptosis-related protein expression were detected by Western blotting. Results After 28 days of perfusion, noaneurysm was formed in the control group while aneurysms were observed in the angiotensin Ⅱ group. The abdominal aortasof mice in the angiotensin Ⅱ group were significantly dilated compared with those in the control group. HE staining indicatedthat the vascular lumen was enlarged and the arterial wall was attenuated in the angiotensin Ⅱ group compared with the controlgroup. Masson and VVG staining showed that the degradation of elastic fibers and the accumulation of AAA-related collagendeposition were significantly increased in the angiotensin Ⅱ group. Immunofluorescence staining indicated that the expressionsof NLRP3, GSDMD, Caspase-1 and IL-18 were increased in the angiotensin Ⅱ group. Western blotting result showed that therewas no significant difference in the expression level of IL-18 protein in MOVAS between these two groups (P > 0.05) ; while theexpression levels of NLRP3, GSDMD and Caspase-1 proteins in the angiotensin Ⅱ group were higher than those in the controlgroup (P < 0.05) . Conclusion The formation of angiotensin Ⅱ-induced AAA is related to pyroptosis, it reducing the numberof vascular smooth muscle cells in the membrane layer and promoting the destruction of the elastic layer of vascular wall via theinflammatory microenvironment, pyroptosis play an important role in AAA occurrence and development.

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