基于环磷酸腺苷/蛋白激酶A/环磷酸腺苷反应元件结合蛋白通路探究丙泊酚对局灶性脑缺血再灌注大鼠神经功能改善机制  

Mechanism of propofol improving neurological function in rats with focal cerebral ischemia-reperfusion based on cAMP/PKA/CREB pathway

作　　者：王岩英[1] 周进国 刘晓宁 王芳[1] 张光信[1] WANG Yanying;ZHOU Jinguo;LIU Xiaoning;WANG Fang;ZHANG Guangxin(Department of Anesthesiology,Handan First Hospital,Handan 056002,China)

机构地区：[1]邯郸市第一医院麻醉科,河北邯郸056002

出　　处：《陕西医学杂志》2024年第4期455-461,共7页Shaanxi Medical Journal

基　　金：河北省邯郸市科学技术研究与发展计划项目(1823208074ZC)。

摘　　要：目的:基于环磷酸腺苷/蛋白激酶A/环磷酸腺苷反应元件结合蛋白(cAMP/PKA/CREB)通路探究丙泊酚对局灶性脑缺血再灌注大鼠神经功能的改善机制。方法:采用改良线栓法缺血2 h,再灌注24 h建立大鼠脑缺血再灌注损伤(CIRI)模型,将造模成功大鼠随机分为模型组和丙泊酚低(1 mg/ml)、中(2.5 mg/ml)、高剂量(5 mg/ml)组,各12只,另设含有12只大鼠的假手术组。分组后即开始给药,1次/d,共4周,末次给药12 h后,采用改良神经功能评分(mNSS)法进行神经缺损评分;采用TTC染色法检测脑梗死面积;HE、Nissl染色进行神经元细胞及尼氏小体形态学观察;Tunel法进行神经元细胞凋亡检测;Elisa法检测脑组织cAMP、脑源性神经营养因子(BDNF)、神经生长因子(NGF)含量;免疫荧光法检测脑组织环磷酸腺苷(cAMP)、p-PKA、p-CREB阳性细胞数及其蛋白共表达阳性细胞数;Western blot法检测脑组织PKA、p-PKA、CREB、p-CREB蛋白表达量。结果:模型组大鼠比较假手术组大鼠的mNSS评分、脑梗死面积百分比显著增加(均P<0.05),HE染色和Nissl染色可见明显的神经元细胞损伤和尼氏小体破坏,脑组织cAMP、BDNF、NGF含量和PKA、p-PKA、CREB、p-CREB蛋白表达量显著下降,模型组大鼠比较假手术组大鼠的cAMP、p-PKA、p-CREB阳性细胞数和蛋白共表达阳性细胞数也明显下降(均P<0.05)。与模型组比较,丙泊酚给药组大鼠mNSS评分、脑梗死面积百分比显著降低(均P<0.05),HE染色和Nissl染色可见神经元细胞损伤和尼氏小体破坏有不同程度改善,脑组织cAMP、BDNF、NGF含量和PKA、p-PKA、CREB、p-CREB蛋白表达量显著升高(均P<0.05),cAMP、p-PKA、p-CREB阳性细胞数及其蛋白共表达阳性细胞数均显著升高(均P<0.05)。结论:丙泊酚可能通过cAMP/PKA/CREB通路改善CIRI大鼠神经功能。Objective:To investigate the mechanism of propofol in improving neurological function in rats with focal cerebral ischemia-reperfusion based on the cAMP/protein kinase A/cAMP response element-binding protein(cAMP/PKA/CREB)pathway.Methods:The rats CIRI model was established by improved thread occlusion for 2 hours and reperfusion for 24 hours.The successful rats were randomly divided into model group,and low(1 mg/ml),medium(2.5 mg/ml)and high dose(5 mg/ml)propofol groups with 12 rats in each group,and 12 rats in sham operation group.After grouping,the drug was administered once a day for 4 weeks.After the last administration for 12 hours,the nerve defect was scored by mNSS method;TTC staining was used to detect the area of cerebral infarction;HE and Nissl staining were used to observe the morphology of neuronal cells and Nissl bodies;TUNEL method was used to detect neuronal apoptosis;the contents of cAMP,BDNF and NGF in brain tissue were detected by ELISA;the number of cAMP,p-PKA,p-CREB positive cells and the number of protein co-expression positive cells were detected by immunofluorescence;the protein expressions of PKA,p-PKA,CREB and p-CREB were detected by Western blot.Results:Compared with the sham group,the mNSS scores,percentage of cerebral infarction area in model group increased significantly(all P<0.05).HE staining and Nissl staining showed obvious neuronal cell damage and Nissl body destruction.The contents of cAMP,BDNF,NGF and the expression of PKA,p-PKA,CREB and p-CREB protein in brain tissue decreased significantly,and the number of cAMP,p-PKA,p-CREB positive cells and their protein co-expression positive cells decreased significantly(all P<0.05).Compared with the model group,the mNSS scores and the percentage of cerebral infarction area in the propofol administration group were significantly lower(all P<0.05).HE staining and Nissl staining showed that the neuronal cell damage and Nissl body destruction were improved in varying degrees,and the contents of cAMP,BDNF,NGF and the expression of PKA,p-PKA,

关 键 词：丙泊酚 局灶性脑缺血再灌注损伤 神经功能 环磷腺苷 蛋白激酶A 环磷腺苷效应元件结合蛋白 

分 类 号：R-332[医药卫生]