H2S通过调控ACE2减轻小鼠颈动脉部分结扎术致动脉粥样硬化程度  

Hydrogen Sulfide Attenuates Atherosclerosis in a Partially Ligated Carotid Artery Mouse model via Regulating Angiotensin Converting Enzyme 2 Expression

作　　者：蔺艳军 曾华甦 高霖[1] 张绘莉[1] LIN Yan-jun;ZENG Hua-su;GAO Lin;ZHANG Hui-li(Department of Cardiology,Shanghai Ninth People's Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200000,China)

机构地区：[1]上海交通大学附属第九人民医院心内科,上海市200011

出　　处：《中国分子心脏病学杂志》2019年第1期2758-2763,共6页Molecular Cardiology of China

基　　金：国家自然科学基金(81570037);上海市科学技术委员会科研计划项目(13ZR1424200)

摘　　要：目的探讨硫化氢(hydrogen sulfide,H_2S)在载脂蛋白E (apolipoproteinE,ApoE)基因敲除小鼠颈动脉部分结扎术(Partial Ligation of left carotid artery,PLA)结合高脂喂养致动脉粥样硬化模型中对血管紧张素转换酶2 (angiotensin converting enzyme 2,ACE2)的调节作用。方法8周龄雄性ApoE-/-小鼠随机分为假手术组,PLA+生理盐水组,PLA+NaHS(硫化氢供体药物,1mg/kg/day)组,PLA+PAG(DL-propargylglycine,10mg/kg/day)组,PLA+NaHS+MLN-4760 (selective ACE 2 inhibitor,0.5mg/kg/day)组。采用左侧颈动脉部分结扎术结合高脂饲料喂养复制小鼠动脉粥样硬化模型,1周或4周后处死小鼠,收集外周血,分离结扎侧颈动脉,检测血浆H_2S活性,HE染色、Masoon染色观察颈动脉病变程度,免疫组化、western blot检测颈动脉组织中ACE2、CSE的表达,Elisa检测颈动脉组织中Angiotensin 1-7 (Ang (1-7)、AngiotensinII(AngII)的表达水平。结果PLA术后4周,小鼠动脉粥样硬化形成。与生理盐水组相比,NaHS干预组ACE2、Ang(1-7)表达水平明显上调,AngII表达水平下降(P<0.05),颈动脉粥样硬化面积明显缩小,内膜增生程度减轻,应用MLN-4760后,NaHS的抗动脉粥样硬化作用被逆转。PAG干预组ACE2、Ang(1-7)表达水平下降,AngII表达水平明显上升(P<0.05),颈动脉内膜增生程度加重,管腔明显狭窄甚至闭塞。结论在颈动脉结扎结合高脂饮食诱导的动脉粥样硬化模型中,硫化氢可通过调控ACE2的表达水平延缓动脉粥样硬化的进展。Objective To evaluate the effect of exogenous Hydrogen Sulfide(H2S) on angiotensin converting enzyme 2(ACE2) in the acute flow disturbance-induced atherosclerosis. Methods Male ApoE-/- mices aged 8 weeks were randomly divided into five groups: sham, PLA+saline, PLA+NaHS( 1mg/kg/day), PLA+PAG( 10mg/kg/day), PLA+NaHS +MLN-4760(0.5mg/kg/day). The mice underwent PLA or sham operation and then were fed a high-fat diet for 4 weeks that contained 10% fat from lard and was supplemented with 2%(w/w) cholesterol. Four weeks postligation, all the mice were sacrificed. Plasma level of H2S was calculated. Samples of carotid arteries and blood were collected and stored at -80℃. The LCA tissues were obtained and stained by HE staining and Masson staining to observe the pathological changes. The expression levels of ACE2 were detected by Western blot and Immunohistochemistry. The expression level of CSE were detected by Western blot. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of AngII and Ang(1-7) in the LCA tissue. Results Four weeks after ligation, NAHS treatment significantly impeded the plaque development, as characterized by alleviated neointimal hyperplasia and less atherosclerotic lesions in LCA. The levels of ACE2 and Ang(1-7) were upregulated, the AngII level was lower than the saline group. On the other hand, inhibition of H2S formation by PAG induced advanced atherosclerosis, as evidenced by more severe neointimal hyperplasia and more obvious narrowness in LCA. The levels of ACE2 and Ang (1-7) were deregulated ,and the level of AngII was higher than saline group. The anti-atherosclerosis property of NaHS was also significantly abolished by treatment with MLN-4760. Conclusion H2S may protect against atherosclerosis in acute flow disturbanceinduced atherosclerosis via upregulating the level of ACE2.

关 键 词：动脉粥样硬化 硫化氢 血管紧张素转化酶2 

分 类 号：R543.5[医药卫生—心血管疾病] R-332[医药卫生—内科学]