Rho激酶抑制剂盐酸法舒地尔对脑缺血再灌注损伤大鼠海马谷氨酸6受体表达及神经细胞凋亡的影响  

Effects of a Rho kinase inhibitor fasudil hydrochloride on GIuR6 and neuronal apoptosis in cerebral ische- mia/reperfusion injury

作　　者：魏秀娥[1] 荣良群[1] 张清秀[1] 王凯[1] 张风玉[1] 

机构地区：[1]徐州医学院第二附属医院神经内科,徐州221006

出　　处：《中华行为医学与脑科学杂志》2013年第9期776-779,共4页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：徐州市科技计划项目（XFIOC030,2010）

摘　　要：目的探讨Rho激酶抑制剂盐酸法舒地尔对大鼠脑缺血再灌注损伤的神经保护作用。方法将清洁级sD大鼠随机分成假手术组、缺血再灌注组、生理盐水组、盐酸法舒地尔组。盐酸法舒地尔组于缺血前30rain腹腔注射盐酸法舒地尔（15mg／kg）进行干预，生理盐水组腹腔注射等量的生理盐水。采用免疫共沉淀和免疫印迹的方法检测盐酸法舒地尔对缺血再灌注6hGluR6的磷酸化水平及蛋白表达的影响；采用TUNEL染色方法检测盐酸法舒地尔对缺血再灌注3d大鼠海马CAl区神经元凋亡的影响。结果1．与假手术组相比，缺血再灌注组6hGluR6丝氨酸的磷酸化水平增高（P〈0．05）；与缺血再灌注组及生理盐水组相比，盐酸法舒地尔组可明显抑制GluR6丝氨酸的磷酸化高峰（P〈0．05）。2．与假手术组大鼠海马CAl区每1mm长度范围内TUNEL阳性细胞数（8．60±2．19）相比，缺血再灌注组3d时TUNEL阳性细胞数（40．20±2．77）增多（P〈0．05）。与缺血再灌注组及生理盐水组（41．80±3．03）相比，盐酸法舒地尔组（19．80±2．86）降低（P〈0．05）。结论盐酸法舒地尔能抑制脑缺血／再灌注诱导的GluR6磷酸化水平，减少海马CAl区神经元的凋亡，对脑缺血再灌注损伤有神经保护作用。Objective To investigate the neuralprotective effect of Rho kinase inhibitor fasudil hydrochlo- ride in cerebral ischemia/reperfusion injury in rats. Methods The SD rats were randomly divided into four groups： the sham group, the ischemia/reperfusion group, the fasudil hydrochloride group and the physiological sa- line group. Fasudil hydrochloride were injected intraperitoneally 30 minutes before ischemia. And the physiological saline group were treated with the intraperitoneal injection of the same volume of saline. The phosphorylation and protein expression of GIuR6 at 6 hours during reperfusion were detected using immunoprecipitation and immunob- lotting analysis to examine the effect of Fasudil hydrochloride. Furthermore, TUNEL staining was used to examine the apoptosis of neurons in rat hippocampal CAI regions after 3 days reperfusion. Results 1. Immunoprecipitati- on and immunoblotting analysis were used to analyze the phosphorylation of GluR6 in serine site. The results showed that the GluR6 serine phosphorylation level increased significantly at 6 h of reperfusion compared with the sham group （P 〈 O. 05 ）. Fasudi｜ hydrochloride group could inhibit the increased phosphorylation of GluR6 at 6 h of reperfusion compared with the ischemia/reperfusion group and saline group, respectively （P 〈 0.05 ）. 2. TUNEL staining was used to examine the apoptosis of neurons in 3 days after reperfusion in CA1 regions of hippocampus. The results indicated that significant numbers of TUNEL positive cells （40.20 ± 2.77） were observed 3 days after ischemia/reperfusion. The numbers of viable neurons per 1 mm length of CA1 pyramidal cells were quantitatively analyzed. Fasudil hydrochloride markedly decreased the neuronal loss compared with the ischemia/reperfusion group （ 19.80 ± 2.86） （P 〈 0.05 ）. Conclusion Fasudil hydrochloride can inhibit induced phosphorylation of GluR6 by the ischemia/reperfusion. Fasudil hydrochloride can reduce the neurons apoptosis in hippocampal CA1 regions, and perform a

关 键 词：盐酸法舒地尔 RHO激酶抑制剂 全脑缺血 谷氨酸6受体 细胞凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]