利拉鲁肽对脑缺血再灌注损伤大鼠脑自由基代谢及血管内皮生长因子表达的影响

liraglutide on superoxide dismutase,malonaldehyde metabolism and the expression of vascular endothelial cell growth factor in Cerebral Ischemia-reperfusion

目的探讨利拉鲁肽对大鼠脑缺血再灌注损伤脑自由基代谢及血管内皮生长因子(VEGF)表达的影响。方法将SD大鼠60只随机分为假手术组、脑缺血再灌注组和利拉鲁肽大、中、小剂量组,每组12只,后4组应用线栓法制备大脑中动脉闭塞2 h后再灌注模型,脑缺血再灌注24 h后对其神经功能缺损进行评分,采用2,3,5-氯化三苯基四氮唑(TTC)染色法检测脑梗死灶大小,取脑组织制备成10%匀浆检测其超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量,采用免疫印迹(Western blot)法及实时荧光定量PCR(Real-time PCR,qPCR)检测大鼠脑组织VEGF的表达。结果与假手术组比较,脑缺血再灌注组脑组织SOD活性降低(95.49±8.16)Mu/L,MDA含量增加(8.15±1.14)μmol/L(P<0.05),脑组织VEGF蛋白表达量增加(0.64±0.18、6.69±0.54)(均P<0.05);与脑缺血再灌注组比较,利拉鲁肽可改善24 h后神经功能缺损(14.45±1.88、12.52±1.49、10.6±1.35)分及减少脑梗死体积(128.1±12.6、105.8±11.7、92.6±10.3)mm^3(均P<0.05),脑组织SOD活性增加(104.89±8.38、112.17±9.35、120.2±10.2)Mu/L,MDA含量减少(7.66±1.23、7.04±1.08、6.49±0.97)μmol/L(均P<0.05),脑组织VEGF蛋白表达量增加[(0.74±0.15、0.84±0.23、0.96±0.24)(8.15±1.56、8.94±1.61、9.89±1.78)](均P<0.05),且与剂量呈正相关性。结论利拉鲁肽能减少大鼠脑缺血再灌注大鼠神经细胞的氧自由基浓度及上调VEGF表达改善脑灌注损伤。

Objective To observe the effect of liraglutide on SOD、MDA metabolism and the expression of VEGF in cerebral ischemia-reperfusion rats. Methods 60 male SD rats were randomly divided into sham group（ n =12）,ischemia-reperfusion group（ n =12）,high-dose liraglutide treatment group（ n =12）,middle-dose liraglutide treatment group（ n =12）and low-dose liraglutide treatment group（ n =12）. The later 4 groups were occluded the middle cerebral artery for 2 hours and reperfused. The 5 groups was sacrificed 24 hours after operation,The neurological dysfunction was scored. The cerebral infarction size was measured with 2,3,5- triphenyltetrazolium chloride staining. and then the level of SOD、MDA were detected,Western blotting method and Real time PCR was used to detect the expression of VEGF. Results Compared with the sham operation group,the activity of SOD of their brain tissue decreased（95.49±8.16）Mu/L,the content of MDA increased（8.15±1.14）μmol/L,the content of VEGF increased （0.64±0.18、6.69±0.54）（ P 〈 0.05 ） in the model group;Compared with I/R group,the infarct volume was decreased （14.45±1.88、12.52±1.49、10.6±1.35）and the neurological function scale was decreased（128.1±12.6、105.8±11.7、92.6±10.3） too. the activity of SOD of their brain tissue increased（104.89±8.38、112.17±9.35、120.2±10.2）Mu/L,the content of MDA decreased（7.66±1.23、7.04±1.08、6.49±0.97）μmol/L,the content of VEGF increased[（0.74±0.15、0.84±0.23、0.96±0.24）（8.15± 1.56 、8.94±1.61、9.89±1.78）] （ P 〈 0.05 ） in the liraglutide pretreatment group. and the positive correlation was showed between the different dose groups. Conclusion liraglutide can protect brain in cerebral ischemia-reperfusion rats through various way,such as decreasing the concentration of hydroxy radical and promoting the expression of VEGF.