法舒地尔保护小鼠脑缺血再灌流损伤的机制  

The neuroprotective effects of Fasudil on brain ischemia/reperfusion injury through improving cerebral blood flow

作　　者：杨锡彤 程建杰 徐弘扬 王光明 YANG Xi - tong;CHENG Jian - jie;XU Hong - yang;WANG Guang - ming(Genetic testing center,the First Affiliated hospital of Dali University,Dali 671000,Yunnan,China)

机构地区：[1]大理大学第一附属医院基因检测中心

出　　处：《广东医学》2019年第11期1521-1525,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81360206);云南省中青年学术技术带头人后备人才基金项目(编号:2014HB025);云南省教育厅科学研究基金项目(编号:2019J0775)

摘　　要：目的探讨法舒地尔(Fasudil)保护小鼠脑缺血/再灌流(ischemia/reperfusion,I/R)损伤的作用机制。方法对成年C57BL/6J雄性小鼠进行法舒地尔灌胃处理,然后行大脑中动脉闭塞(middle cerebral artery occlusion,MCAo)动物模型,用氯化三苯四氮唑染色法(2,3,5-triphenyltetrazolium chloride,TTC)染色以分析脑梗死情况,激光散斑(LaserSpeckle)图像系统分析脑血流改变情况;对成年C57BL/6J雄性小鼠进行法舒地尔灌胃处理后,取脑组织用免疫印迹分析过氧化物酶体激活受体α(peroxisome proliferators-activated receptoralpha,PPARα)的表达状态,实时荧光定量PCR检测超氧化物歧化酶(superoxide dismutase,SOD)1~3的表达及化学发光法检测SODs的活性。结果雄性C57BL/6J小鼠经法舒地尔处理后,脑组织中PPARα和SOD1、SOD2、SOD3的表达升高(P=0.000、0.000、0.000、0.000),SODs酶活性升高(P=0.000);在小鼠脑MCAO动物模型中,法舒地尔预处理使脑坏死体积明显减小(P=0.000),再灌流后脑坏死区域的脑血流强于羟甲基纤维素(carboxymethyl cellulose,CMC)处理小鼠。结论法舒地尔通过促进PPARα的表达,影响SOD1、SOD2、SOD3的表达及活性,以改善小鼠脑缺血区域的脑血流,从而保护I/R损伤。Objective To investigate the neuroprotective mechanisms of Fasudil on brain ischemia /reperfusion injury with MCAO model.Methods Separate C57BL/6J mice was treated with Fasudil ( 10 mg /kg.BW) through intra- gastric administration for 3 days,while the mice treated with CMC ( 10 mL 0. 5% CMC/kg.BW) were used as control group.The treated mice were subjected to 60 minutes of focal ischemia and 18 hours reperfusion.The brain was collected and the infarct volume was analyzed through TTC staining with MCID image system.Cerebral blood flow was measured with laser speckle in MCAO.The brain collected from the mice treated with Fasudil or CMC,without MCAO,was used to check the expression of PPAR - alpha,SOD1,2,3 and the activity of SODs.The expression of PPAR - alpha was checked with immuno - blotting assay,the mRNA of SOD1,2,3 was assayed with real - time PCR,and the activity of SODs was analyzed with a chemiluminescence method.Results Compared with control group,the PPAR - alpha and SOD1,2,3 in the brain from Fasudil treated mice was significantly higher ( P = 0. 000).The activity of SODs in the Fa- sudil treated mice was significantly higher than control group ( P = 0. 000).The infarct volume was significantly attenuated with application of Fasudil ( P = 0. 000).Compared with the control group, the cerebral blood flow was attenuated with application of Fasudil.Conclusion The neuroprotective mechanisms of Fasudil on the brain ischemia /reperfusion injury is through promoting the expression of PPAR - alpha to boost the expression of SOD1,2,3 and the activity of SODs to en- hance the cerebral blood flow to attenuate the I/R injury.

关 键 词：脑缺血 法舒地尔 脑血流 过氧化物酶体激活受体α 超氧化物歧化酶 

分 类 号：R743.31[医药卫生—神经病学与精神病学] R322.81[医药卫生—临床医学]