尤瑞克林对局灶性脑梗死大鼠Slit2/Robo1表达的影响

Expressions of Slit2/Robo1 in Rats with Cerebral Infarction and Changes after Intervened by Urinary Kallidinogenase

目的:研究神经导向因子Slit2及其受体Robo1在大鼠脑梗死模型和尤瑞克林干预后的缺血脑组织中的表达变化。方法:54只雄性SD大鼠随机分为假手术组、模型组和尤瑞克林组,再随机分为梗死后缺血1 d、3 d、7 d三个亚组,每组各6只。采用线栓法制备大鼠永久性大脑中动脉缺血(permanent middle cerebral artery occlusion,pMCAO)模型。尤瑞克林组给予尤瑞克林尾静脉注射,模型组给予等体积生理盐水。应用Westernblot方法检测不同时间点Slit2蛋白和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)在缺血脑组织中的表达情况;RT-PCR方法检测不同时间点Robo1 mRNA的表达情况。结果:与假手术组相比,模型组和尤瑞克林组Slit2蛋白和eNOS蛋白的表达在各时间点均明显增高;尤瑞克林组与模型组相比,两蛋白在各时间点均呈现高表达,差异有统计学意义(P<0.05)。且Slit2蛋白和eNOS蛋白的表达变化呈显著正相关(P<0.01)。Robo1在模型组和尤瑞克林组中表达较假手术组早期明显降低,在3 d开始上升,呈升高趋势,7 d仍未达到假手术组大鼠的表达水平(P<0.05)。结论:局灶性脑梗死后大鼠Slit2蛋白表达升高,且Slit2可能参与大鼠脑梗死后血管新生过程,尤瑞克林可能通过升高Slit2/Robo1促进大鼠脑梗死后的血管新生过程。

Objective:To explore the relationship between Slit2/Robo1 signaling and investigate the expression of Slit2&Robo1 in rat cerebral infarction models and Urinary Kallidinogenase(UK)intervention.Methods:Fifty-four male Sprague Dawley rats were assigned into sham group(SG),model group(MG)and Urinary Kallidinogenase group(UKG)randomly.The groups were further randomized into three subgroups:1 d,3 d,and7 d after infarction occurred.Cerebral ischemia model was established via the suture method.UKG were injected UK via tail vein and the MG received equal volume of saline.Western-blot was used to analyze the level of Slit2 and eNOS at different time points and RT-PCR was used to detect the expression of Robo1 mRNA.Results:The expression of Slit2 and eNOS was significantly increased in the MG and UKG,meanwhile,it was positively correlated with eNOS.In the MG and UKG,the levels of Robo1 were significantly decreased in the early period and the expression remained suppressed until 3 d after occlusion,not yet reached the level of SG by 7 d,the difference was statistically significant.Conclusions:The expression of Slit2 protein in rats after focal cerebral infarction is increased,and Slit2 may participate in the process of angiogenesis after cerebral infarction in rats.Urinary Kallidinogenase may promote the process of angiogenesis in rats after cerebral infarction by increasing Slit2/Robo1.