血管紧张素II经酸性鞘磷脂酶/神经酰胺通路致动脉内皮功能障碍的作用  

Acid sphingomyelinase/ceramide mediates arterial angiotensin Ⅱ-induced endothelial dysfunction

作　　者：苏玉婷[1] 孟星星 程耀萍 谢小萍[1] 张海军[1] 常耀明[1] 暴军香[1] SU Yu-ting MENG Xing-xing CHENG Yao-ping XIE Xiao-ping ZHANG Hai-jun CHANG Yao-ming BAO Jun-xiang(Department of Aerospace Hygiene, Fourth Military Medical University, Xi ＇an 710032, Shaanxi, China)

机构地区：[1]第四军医大学航空航天卫生学教研室,陕西西安710032

出　　处：《心脏杂志》2017年第1期34-39,共6页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(31071045;81401550)

摘　　要：目的探讨酸性鞘磷脂酶(acid sphingomyelinase,ASM)/神经酰胺(ceramide,Cer)通路在血管紧张素II(angiotensin II,Ang II)致动脉内皮功能障碍中的作用和机制。方法针对对照组,低、中、高剂量AngⅡ组及ASM抑制剂地昔帕明(desipramine,Dpm)组,采用Western blot检测法、实时定量PCR和免疫荧光化学方法检测ASM、Cer、总内皮型一氧化氮合酶(total endothelial nitric oxide synthase,t-e NOS)和磷酸化e NOS(phosphorylated e NOS,p-e NOS)含量变化;采用血管环张力描记技术检测主动脉血管环的舒张功能;采用二氢乙啶(dihydroethidium,DHE)荧光探针检测血管环超氧阴离子(superoxide anion,O_2^-·)水平。结果与对照组相比,AngⅡ孵育后主动脉对乙酰胆碱(acetylcholine,Ach)的舒张反应显著降低(P<0.05),呈剂量依赖性,对硝普钠(sodium nitroprusside,SNP)的舒张反应无明显改变;AngⅡ孵育组动脉ASM表达显著增加(P<0.05),Cer含量显著增多(P<0.05);ASM抑制剂Dpm可显著抑制AngⅡ所致ASM和Cer改变(P<0.05),亦可使动脉对Ach的舒张反应恢复正常;AngⅡ可显著降低动脉p-e NOS含量(P<0.05),增加O_2^-·水平(P<0.05),Dpm则可使p-e NOS增加(P<0.05),降低O_2^-·(P<0.05)。结论 ASM/Cer可能通过抑制e NOS磷酸化并增加O_2^-·含量介导AngⅡI所致动脉内皮功能障碍。AIM To investigate the effect and mechanism of acid sphingomyelinase （ASM）/ceramide （Cer） pathway in arterial endothelial dysfunction caused by augiotensin II （Ang II）. METHODS Western blot, real-time polymerase chain reaction （RT-PCR） and immunohistochemistry were carried out to examine the content of ASM, Cer, total endothelial nitric oxide synthase （t-eNOS） and phosphorylated eNOS （p-eNOS）. Isometric force recording system was used to detect vascular function. Dihydroethidium （DHE） fluorescent probe was used to evaluate the level of superoxide anion （O2^-·） in arteries. RESULTS After incubation with AngII （ 100 nmol/L）, the vasodilation response of aorta to acetylcholine （Ach） decreased significantly （P 〈 0. 05 ）, which was dose dependent, whereas vasodilation response to sodium nitroprusside （SNP） did not change. ASM protein and Cer level were increased significantly by Ang II incubation compared with the control level （P 〈 0. 05 ）, which could be reduced by incubation with desipramine （Dpm）, a specific ASM inhibitor. Dpm pretreatment significantly increased the vasodilation response to Ach. AnglI could significantly reduce p-eNOS content of artery as well as increase the level of O2^-· , whereas Dpm could increase p-eNOS and lower the level of O2^-· . CONCLUSION Ang II induces endothelial dysfunction through ASM/Cer, which might inhibit the phosphorylation of eNOS and enhance the content of O2^-·.

关 键 词：血管紧张素II 动脉 内皮 酸性鞘磷脂酶 神经酰胺 

分 类 号：R318.01[医药卫生—生物医学工程]