【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。更多还原

【Abstract】 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,CREB and p-CREB proteins in brain tissue were significantly higher, the number of cAMP,p-PKA,p-CREB positive cells and their protein co-expression positive cells increased significantly(all P<0.05).Conclusion:Propofol may improve the neurological function of CIRI rats through cAMP/PKA/CREB pathway.更多还原