凝血酶预处理对海马神经元的保护作用  

The Effect of Thrombin Preconditioning on the Apoptosis of Hippocampal Neurons

作　　者：杨文琼[1] 孙圣刚[2] 童萼塘[2] 

机构地区：[1]郧阳医学院附属东风医院神经内科,湖北十堰442008 [2]华中科技大学同济医学院附属协和医院神经内科,湖北武汉430022

出　　处：《郧阳医学院学报》2006年第6期325-327,330,共4页Journal of Yunyang Medical College

基　　金：海南省自然基金资助项目(N30215)

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48h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Bc l-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(P<0.01),Bc l-2表达上调,Bax表达明显下调(P<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(P<0.01),Bc l-2表达上调,Bax表达明显下调(P<0.01)。TRAP预处理组与TPC组相比差异无显著性(P>0.05)。结论:TPC可抑制TM导致的神经细胞凋亡,激活PAR-1,促进Bc l-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。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 5 μmol/ L thrombin receptor activating peptides （TRAP） for 48h before exposed to 40U/ml TM, respectively. The number of apoptotic ceUs 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（P 〈0.01 ）. Bcl -2 protein expression was increased, Bax protein expression was decreased significantly in TPC group（P 〈0.01 ）. Compared to Sham group, the number of apoptotic ceUs and apoptotic rate of neurons were decreased sig- nificantly in TRAP group（ P 〈 0.01 ）. Bcl - 2 protein expression was increased, Bax protein expression was decreased signif- icantly in TRAP group（P 〈0.01 ）. There was no significant difference in TRAP group and TPC group（P 〉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.

关 键 词：凝血酶预处理 细胞凋亡 BCL-2 BAX 蛋白酶激活受体-1 

分 类 号：R363[医药卫生—病理学]