血管扩张刺激磷蛋白缓解脓毒症小鼠心肌细胞受损  

Vasodilator-Stimulated Phosphoprotein Alleviates Cardiomyocyte Damage in Septic Mice

作　　者：张继龙[1] 单萍[1] 韩杨[1] 朱娜 ZHANG Jilong;SHAN Ping;HAN Yang(The First Hospital of Wuhan,Hubei Wuhan 430022,China)

机构地区：[1]湖北省武汉市第一医院急诊科,湖北武汉430022

出　　处：《河北医学》2023年第5期742-747,共6页Hebei Medicine

基　　金：湖北省武汉市卫健委课题,(编号:WX19C40)。

摘　　要：目的:验证二丁酰环磷腺苷(Bucladesine)介导的血管扩张刺激磷蛋白(VASP)变化与脓毒症小鼠心肌细胞作用关系。方法:盲肠结扎穿孔术构建脓毒症小鼠模型,实验分为:对照组(Con)、假手术组(Sham)、模型组(Mod)和Bucladesine干预组(Buc)。生化检测血清和心肌组织中丙二醛(MDA)和超氧化物歧化酶(SOD)含量。流式细胞术检测心肌组织中活性氧(ROS)含量和细胞中线粒体膜电位。TUNEL染色检测心肌细胞凋亡。苏木精-伊红(HE)染色观察心肌组织病变。Western blot检测心肌组织中血管扩张刺激磷蛋白磷酸化表达水平。结果:盲肠结扎穿孔成功诱导脓毒症小鼠模型,与对照组和假手术组相比,模型组血清和心肌组织中MDA含量显著升高(P<0.05),SOD含量显著降低(P<0.05),ROS含量显著升高(P<0.05),p-VASP和VASP蛋白表达水平显著升高(P<0.05),心肌纤维断裂,疏松,间隙增大,TUNEL棕色面积着色增加。与模型组相比,Buc组血清和心肌组织中MDA含量显著降低(P<0.05),SOD含量显著升高(P<0.05),ROS含量显著降低(P<0.05),p-VASP和VASP蛋白表达水平显著降低(P<0.05),心肌纤维紧密,TUNEL棕色面积着色减少。结论:Bucladesine抑制p-VASP能够缓解脓毒症导致的心肌功能抑制,其作用机理与调节心肌细胞线粒体活性和增强细胞抗氧化能力相关。Objective:Validation of dibutyryl cyclic adenosine(Bucladesine)-mediated changes in vasodilator-stimulated phosphoprotein(VASP)in relation to cardiomyocyte action in septic mice.Methods:A mouse model of sepsis was constructed by cecum ligation perforation,and the experiment was divided into:control(Con),sham-operated(Sham),model(Mod)and Bucladesine(Buc)groups.Biochemical assays were performed to detect tissue malondialdehyde(MDA)and superoxide dismutase(SOD)levels in serum and myocardium.Flow cytometry was performed to detect reactive oxygen species(ROS)levels in myocardial tissue and mitochondrial membrane potential in cells.TUNEL staining was performed to detect myocardial tissue apoptosis.Hematoxylin-eosin(HE)staining was performed to observe myocardial lesions.p-VASP expression levels in myocardial tissues were measured by Western blot.Results:Compared with the control and sham groups,the model group showed significantly higher MDA levels(P<0.05),lower SOD levels(P<0.05),higher ROS levels(P<0.05),higher p-VASP and VASP protein expression levels(P<0.05)in serum and myocardial tissue,myocardial fiber breakage,laxity,increased gap,and increased TUNEL brown area coloring.Compared with the model group,the serum and myocardial tissue in the Buc group showed significantly lower MDA content(P<0.05),higher SOD content(P<0.05),lower ROS content(P<0.05),lower p-VASP and VASP protein expression levels(P<0.05),tighter myocardial fibers,and reduced TUNEL brown area coloration.Conclusion:Inhibition of p-VASP by Bucladesine can alleviate the myocardial dysfunction caused by sepsis,which mechanism of action is associated with modulation of cardiomyocyte mitochondrial activity and enhancement of cellular antioxidant capacity.

关 键 词：血管扩张刺激磷蛋白 脓毒症 心肌功能 

分 类 号：R459.7[医药卫生—急诊医学]