凝血酶预处理对海马神经元保护作用.docVIP

凝血酶预处理对海马神经元保护作用 　 作者：杨文琼,孙圣刚,童萼塘 　[摘要]　　目的:研究凝血酶预处理（TPC）对大剂量凝血酶（TM）诱导海马神经元凋亡的影响，阐明TPC的保护作用。方法:海马神经元分别经生理盐水（Sham组）、小剂量TM（TPC组）及凝血酶受体激活肽（TRAP组）预处理48 h后再加大剂量TM作用24 h。TUNEL法及流式细胞术检测凋亡细胞数和凋亡百分率，应用免疫细胞化学方法检测神经元Bcl-2及Bax蛋白表达。结果:与Sham组比较，TPC组TUNEL阳性细胞数和凋亡百分率明显降低（Plt;0.01），Bcl-2表达上调，Bax表达明显下调（Plt;0.01）。与Sham组比较，TRAP预处理组TUNEL阳性细胞数和凋亡百分率明显降低（Plt;0.01），Bcl-2表达上调，Bax表达明显下调（Plt;0.01）。TRAP预处理组与TPC组相比差异无显著性（Pgt;0.05）。结论:TPC可抑制TM导致的神经细胞凋亡，激活PAR-1，促进Bcl-2的表达和降低Bax的表达，可能是TPC抑制细胞凋亡的机制之一。 　　[关键词]　　凝血酶预处理；细胞凋亡；Bcl-2；Bax；蛋白酶激活受体-1 　　The Effect of Thrombin Preconditioning on the Apoptosis of Hippocampal Neurons 　　 　　Abstract:Objective To investigate the effect of thrombin preconditioning（TPC）on the apoptosis of hippocampal neurons and explore the potential mechanisms. Methods Hippocampal neurons were pretreated with saline,1U/ml TM and 5μmol/L thrombin receptor activating peptides（TRAP）for 48h before exposed to 40U/ml TM,respectively.The number of apoptotic cells and apoptotic rate of neurons were measured by terminal deoxynucleotidyl transferase (TdT) mediated dUTP-biotin nick end-labeling（TUNEL）method and Flow cytometry.Bcl-2 and Bax protein expression were measured by immuocytochemical method.Results The number of apoptotic cells and apoptotic rate of neurons were decreased significantly in TPC group（Plt;0.01）.Bcl-2 protein expression was increased, Bax protein expression was decreased significantly in TPC group（Plt;0.01）.Compared to Sham group,the number of apoptotic cells and apoptotic rate of neurons were decreased significantly in TRAP group（Plt;0.01）.Bcl-2 protein expression was increased,Bax protein expression was decreased significantly in TRAP group（Plt;0.01）.There was no significant difference in TRAP group and TPC group（Pgt;0.05）.Conclusion TPC could inhibit neuron apoptosis induced by TM.The mechanism of neuroprotective effect of TPC may be through activating PAR-1,up-regulating Bcl-2 protein expression and down-regulating Bax expression. 　　Key words:Thrombin preconditioning