辛伐他汀预防小鼠高脂血症合并脑缺血损伤及其对时钟基因的调控  

Simvastatin prevents cerebral ischemia-reperfusion injury and regulates clock genes in hyperlipidemia mice

作　　者：王敏[1] 阙文轩 陈刚领[2] WANG Min;QUE Wenxuan;CHEN Gangling(Medical Outpatient Department,China Pharmaceutical University,Nanjing 211198,China;School of Traditional Chinese Medicine,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学医学门诊部,江苏南京211198 [2]中国药科大学中药学院,江苏南京211198

出　　处：《中国药理学与毒理学杂志》2024年第7期496-503,共8页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(82074058);中央高校基本科研业务费专项资金资助(2632023GR03)。

摘　　要：目的探讨辛伐他汀对小鼠高血脂合并脑缺血再灌注损伤(CIRI)的预防作用及机制。方法将60只C57BL/6J小鼠分为假手术组、CIRI组〔采用大脑中动脉阻塞再灌注法(MCAO/R)制备小鼠CIRI模型〕、高脂组(ip给予泊洛沙姆407)、高脂+CIRI组(ip给予泊洛沙姆407,24 h后MCAO/R造模)、高脂+CIRI+辛伐他汀5和10 mg·kg^(-1)组(先ig给予辛伐他汀连续7 d,后按照高脂+CIRI组处理)。再灌注24 h后,对各组小鼠进行神经功能缺损评分;转棒实验测定掉落潜伏期;自主活动测试小鼠活动次数;TTC染色测定脑梗死体积;激光散斑血流成像检测小鼠脑血流量;取全血分离血清,并测定总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-PX)含量;实时荧光定量PCR和Westen印迹法检测小鼠右脑皮质时钟基因Rev-erbα和Bmal1 mRNA及蛋白表达水平。结果与假手术组相比,CIRI组小鼠神经功能缺损评分升高(P<0.01),掉落潜伏期缩短(P<0.01),活动次数减少(P<0.01)。与CIRI组相比,高脂+CIRI组小鼠神经功能损伤加重(P<0.01);脑梗死体积显著增加(P<0.01);脑血流量显著降低(P<0.01);血清中TC,TG,LDL-C及MDA含量显著升高(P<0.01),GSH-PX水平显著降低(P<0.01);脑皮质Rev-erbαmRNA和蛋白表达水平显著下调(P<0.01);Bmal1 mRNA和蛋白表达水平显著上调(P<0.01)。与高脂+CIRI组相比,高脂+CIRI+辛伐他汀5和10 mg·kg^(-1)组小鼠神经功能损伤减轻(P<0.01);脑梗死体积显著减小(P<0.01);脑血流量显著增加(P<0.01);血清中TC,TG,LDL-C及MDA含量显著降低(P<0.01),GSH-PX水平显著升高(P<0.01);脑皮质Rev-erbαmRNA和蛋白表达水平显著上调(P<0.01);Bmal1 mRNA和蛋白表达水平显著下调(P<0.01)。结论辛伐他汀预防性给药能有效减轻小鼠高脂血症合并CIRI,其机制与其降血脂及调节时钟基因表达有关。OBJECTIVE To investigate the preventive effect and mechanism of simvastatin on cerebral ischemia-reperfusion injury(CIRI)in hyperlipidemic mice.METHODS Sixty C57BL/6J mice were randomly divided into Sham group,CIRI group(CIRI model was prepared by middle cerebral artery occlusion and reperfusion(MCAO/R)),hyperlipemia group(i.p poloxamer 407),hyperlipemia+CIRI group(i.p poloxamer 407,followed by MCAO/R operation after 24 h),and hyperlipemia+CIRI+simvastatin 5 and 10 mg·kg^(-1) groups(i.g simvastatin for 7 d,and then treated as the hyperlipemia+CIRI group).After reperfusion for 24 h,the neurological deficit score(NDS)was evaluated;the Rotarod experiment was used to determine the first drop latency;autonomous activity was used to test the horizontal and vertical movement frequency of mice;TTC staining was used to measure the volume of cerebral infarction;laser speckle flow imaging was used to detect cerebral blood flow in mice;the kits were used to detect total cholesterol(TC),triglycerides(TG),low-density lipoprotein cholesterol(LDL-C),malondialdehyde(MDA)levels,and glutathione peroxidase(GSH-PX)levels in serum;qPCR and Western blotting were used to detect the mRNA and protein expression levels of clock genes Rev-erbαand Bmal1 in the right cerebral cortex of mice,respectively.RESULTS Compared with the Sham group,the NDS of the CIRI group was significantly increased(P<0.01),the latency period was shortened(P<0.01),and the number of activities was reduced(P<0.01);Compared with the CIRI group,the hyperlipemia+CIRI group showed aggravated neurological damage(P<0.01);the volume of cerebral infarction was significantly increased(P<0.01);the cerebral blood flow was significantly decreased(P<0.01);the levels of TC,TG,LDL-C,and MDA in serum were significantly increased(P<0.01),while the level of GSH-PX was significantly decreased(P<0.01);the mRNA and protein expression levels of Rev-erbα in the cerebral cortex were significantly downregulated(P<0.01);the mRNA and protein expression levels of Bmal1 were significantly upre

关 键 词：辛伐他汀 高血脂 脑缺血再灌注损伤 时钟基因 Rev-erbα BMAL1 

分 类 号：R972[医药卫生—药品]