锰超氧化物歧化酶对氯化钙诱导的小鼠腹主动脉瘤形成的影响及机制  

Effect of Mn-SOD on formation of abdominal aortic aneurysm induced by calcium chlorine in mice and its mechanism

作　　者：陈娇 张蕾 郑飞 王露 晏誉文 郭凌郧 杨建业 沈俊 王家宁 CHEN Jiao;ZHANG Lei;ZHENG Fei;WANG Lu;YAN Yu-wen;GUO Lingyun;YANG Jian-ye;SHEN Jun;WANG Jia-ning(Department of Cardiology,Renmin Hospital,Hubei University of Medicine,Shiyan 442000,China;Institute of Clinical Medicine,Renmin Hospital,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院附属人民医院心内科,湖北十堰442000 [2]湖北医药学院附属人民医院临床研究所,湖北十堰442000

出　　处：《中国病理生理杂志》2022年第10期1773-1780,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81270221);湖北省教育厅中青年人才项目(No.Q20212102)。

摘　　要：目的:初步探讨锰超氧化物歧化酶(manganese superoxide dismutase,MnSOD/SOD2)在小鼠腹主动脉瘤(abdominal aortic aneurysm,AAA)发生发展中的作用及机制。方法:(1)C57BL小鼠(共120只)随机分为3组:正常对照、AAA组和Ad-SOD2组。除对照组外,其余小鼠均通过氯化钙(calcium chloride,CaCl_(2))诱导建成AAA模型,Ad-SOD2组小鼠在建模后腹腔注射Ad-SOD2,并于注射后1、2、4、6周取材,行Western blot检测、HE和弹力纤维染色及ELISA试剂盒检测血清炎症因子等。(2)有/无腺病毒介导的SOD2处理大鼠主动脉血管平滑肌细胞(rat aortic smooth muscle cells,RASMCs)后予以H_(2)O_(2)处理,采用Western blot检测细胞SOD2等蛋白的表达,采用相应的试剂盒检测ROS水平、超氧阴离子水平及SOD2和总SOD活性,采用JC-10染色检测线粒体膜电位。结果:(1)建模成功后,AAA组SOD2和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的表达与对照组相比下降(P<0.05),但在第6周又有所升高(P<0.05),但骨桥蛋白(osteopontin,OPN)表达在2组间无明显差异(P>0.05);AAA组与对照组相比腹主动脉局部炎症反应加重,弹力纤维破坏明显(P<0.01);与AAA组相比,Ad-SOD2组SOD2和α-SMA的表达升高(P<0.01),腹主动脉炎症反应减轻,弹力纤维断裂减少(P<0.01)。各组小鼠血清中炎症因子(IL-6、IL-10、IL-1β)及基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)水平无明显差异(P>0.05)。但血清总SOD活性在建模4周后开始下降,SOD2活性在建模1周最高随后下降(P<0.05)。第6周时,Ad-SOD2组SOD2和SOD活性较AAA组升高(P<0.01)。(2)活性氧(reactive oxygen species,ROS)及超氧阴离子水平检测结果均显示Ad-SOD2组较模型组水平降低(P<0.01),同时JC-10染色提示Ad-SOD2可延缓线粒体膜电位下降(P<0.01)。结论:SOD2可通过清除ROS、延缓线粒体膜电位下降而明显抑制氧化应激条件下H_(2)O_(2)诱导的RASMCs凋亡。同时Ad-SOD2转染可减轻主动脉周围炎症反应、减少弹力纤�AIM:To investigate the role and mechanism of manganese superoxide dismutase,(MnSOD/SOD2)on the pathogenesis and development of AAA in mice.METH_(2)O_(2)DS:(1)C57BL mice(120 mice in total)were randomly divided into 3 groups:control group,AAA group and Ad-SOD2 group.In the model of AAA group,mice were induced by calcium chloride(CaCl_(2)),and the mice in the Ad-SOD2 group were injected with Ad-SOD2 intraperitoneally after modeling.The samples were collected at 1,2,4 and 6 week after the operation,and were subjected to Western blot,HE and elastic fiber staining.ELISA kits were used to detect serum inflammatory factors,etc.(2)Rat aortic smooth muscle cells(RASMCs)were treated with or without adenovirus-mediated SOD2(Ad-SOD2)and then treated with H_(2)O_(2).Western blot was used to detect SOD2 and other protein expressions.The level of ROS or superoxide anion was detected by the corresponding kit,and the mitochondrial membrane potential was detected by JC-10 staining.RESULTS:(1)After the AAA model is successfully established,the expression levels of SOD2 andα-smooth muscle actin(α-SMA)in the AAA group were decreased when compared with those in the control group(P<0.05),while both SOD2 andα-SMA were increased at the 6th week(P<0.05).No significant difference was observed between control and AAA groups on the expression of osteopontin(OPN).Compared with the control group,the local inflammation of abdominal aorta in the AAA group was aggravated,and the elastic fibers were damaged significantly(P<0.01).Compared with the AAA group,the expression levels of SOD2 andα-SMA in the Ad-SOD2 group were increased(P<0.01),meanwhile the inflammation of abdominal aortic and elastic fiber rupture were reduced(P<0.01).Serum inflammatory factors(IL-6,IL-10 and IL-1β)and matrix metalloproteinase 9(MMP9)between groups showed no significant changes(P>0.05).However,the total SOD activity in serum of calcium chloride induced-mice appeared to decrease at the 4th week,while the SOD2 activity reached the peak in the 1st week and then decrea

关 键 词：腹主动脉瘤 发病机制 平滑肌细胞 超氧化物歧化酶 

分 类 号：R543.16[医药卫生—心血管疾病] R363.21[医药卫生—内科学]