Sestrin2在慢性肾功能不全小鼠动脉粥样硬化血栓形成中的作用和机制  

The role and mechanism of Sestrin2 in atherosclerotic thrombosis in mice with chronic renal insufficiency

作　　者：靳文[1] 鄢文[2] 李冬义[1] 劳钰 刘一炫[1] 陈洁琼[1] 赵雅红[1] JIN Wen;YAN Wen;LI Dongyi;LAO Yu;LIU Yixuan;CHEN Jieqiong;ZHAO Yahong(The Third Department of Cardiovascular Medicine,Guangdong Second People's Hospital,Guangzhou 510317,China)

机构地区：[1]广东省第二人民医院心血管内三科,广州510317 [2]广东省第二人民医院肿瘤一科,广州510317

出　　处：《实用医学杂志》2019年第15期2370-2374,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目（编号：81673949）;海珠区科普计划项目（编号：海科工商信计2018-60）

摘　　要：目的探讨Sestrin2在慢性肾功能不全诱导的动脉粥样硬化血栓形成中的作用和分子机制。方法在体实验选用30周龄载脂蛋白E(apolipoprotein E,ApoE)基因敲除小鼠为研究对象,部分予以行5/6肾大部分切除术制备慢性肾功能不全合并动脉粥样硬化小鼠模型,部分予以行假手术。实验分为3组:对照组(行假手术+等容量生理盐水灌胃,n=8)、慢性肾功能不全组(行5/6肾大部分切除术+等容量生理盐水灌胃,n=8)和Sestrin2激动剂组(行5/6肾大部分切除术+10 mg/kg Eupatilin灌胃1周,n=8)。1周后检测FeCl3诱导的颈动脉血栓形成时间和小鼠断尾出血时间;分离小鼠血小板和血浆,流式细胞技术检测血小板内活性氧和P-selectin的表达,蛋白印迹法检测血小板Sestrin2的表达,采用脲酶波氏比色法和苦味酸速率比色法检测小鼠血浆肌酐和尿素氮的表达。结果与对照组相比,慢性肾功能不全小鼠血浆尿素氮和血肌酐水平、血小板活性氧生成以及P-selectin表达显著升高(P <0.05),颈动脉血栓形成时间较对照组明显减少(P <0.05),鼠尾出血时间无显著延长(P> 0.05)。Sestrin2激动剂能够降低慢性肾功能不全小鼠的血浆尿素氮和肌酐水平、血小板活性氧生成以及P-selectin表达(P <0.05),延长颈动脉血栓形成时间(P <0.05),对鼠尾出血时间无明显影响(P> 0.05)。结论 Sestrin2参与了慢性肾功能不全合并动脉粥样硬化性血栓的形成,其机制可能与血小板活化有关。Objective To investigate the role and molecular mechanism of Sestrin2 in atherosclerotic thrombosis induced by chronic renal insufficiency. Methods Apolipoprotein E(ApoE)gene knockout mice of 30 weeks old were selected as the research objects in vivo. Some mice were subjected to 5/6 partial nephrectomy to prepare chronic renal insufficiency model,and some were subjected to sham operation. The experiment was divided into three groups:control group(sham operation + isovolumic saline intragastric administration,n = 8),chronic renal insufficiency group(5/6 partial nephrectomy with isovolumic saline intragastric administration,n = 8)and Sestrin2 agonist group(5/6 partial nephrectomy+10 mg/kg Eupatilinintragastric administration for 1 week,n = 8).One week later,the time of carotid artery thrombosis induced by FeCl3 and the time of tail-cutting bleeding in mice were measured,the platelets and plasma were isolated,the expression of ROS and P-selectin in platelets was detected by flow cytometry,the expression of Sestrin2 in platelets was detected by Western blotting,the expression of creatinine and urea nitrogen in plasma was detected by urease Bohr colorimetry and picric rate colorimetry.Results Compared with the control group,plasma urea nitrogen and creatinine levels,platelet reactive oxygen species(ROS)production and P-selectin expression in chronic renal insufficiency mice increased significantly(P <0.05),carotid artery thrombosis time decreased significantly(P < 0.05),and Tail-cutting hemorrhage time did not prolong significantly(P > 0.05). Sestrin2 agonist could reduce the levels of plasma urea nitrogen and creatinine,the production of platelet reactive oxygen species and the expression of P-selectin in mice with chronic renal insufficiency(P < 0.05),prolong the time of carotid artery thrombosis(P < 0.05),and had no significant effect on the time of tail-cutting bleeding(P > 0.05). Conclusion Sestrin2 participates in the pathological process of atherosclerotic thrombosis induced by chronic renal insufficiency. The

关 键 词：Sestrin2 慢性肾功能不全 动脉粥样硬化 血小板 血栓 

分 类 号：R543.5[医药卫生—心血管疾病]