摘要
目的探讨脑缺血再灌注海马组织腺苷脱氨酶与正常海马组织腺苷脱氨酶性质的差别。方法以测氨法、考马斯蓝染料法、caiyongSDS-PAGE、聚丙烯酰胺凝胶电泳、DNS-Cl法测定再灌注海马组织腺苷脱氨酶与正常海马组织腺苷脱氨酶的酶活性、蛋白含量、分子量测、N-末端氨基酸。结果正常大鼠及假手术大鼠脑海马组织、再灌注脑海马组织ADA的热稳定性、分子量、等电点、N-末端氨基酸、免疫原性及对巯基修饰的敏感性性质基本相似。再灌注脑海马组织巯基修饰的敏感性、热稳定性、酶活性则与正常大鼠及假手术大鼠脑海马组织ADA不同。正常大鼠及假手术大鼠脑海马组织ADA在低浓度PCMB时即被强烈抑制,DTT可使被抑制的酶活性得到部分恢复,说明巯基是正常大鼠.假手术大鼠海马组织ADA活性中心必需的基团。而脑缺血再灌注海马组织ADA在低浓度PC-MB活性无明明显变化,这说明再灌海马组织ADA中巯基不是其活性中心的必需基团。热稳定性测定结果表明脑缺血再灌注海马组织ADA热稳定性较正常大鼠.假手术大鼠海马组织ADA热稳定性强。脑缺血再灌注海马组织ADA活性较正常大鼠。假手术大鼠海马组织ADA活性增强。ADA活性增强可能与热稳定性提高和活化基团不同有关。结论脑缺血再灌注海马组织腺苷脱氨酶性质发生了改变,这些ADA酶性质的变化可能影响腺苷在脑缺血再灌注损伤中的作用。
Objective To study the change of Adenosine Deaminase（ADA） in Hippocampus tissue after reperfusion comparing with those in normal Hippocampus tissue. Methods the molecular weights, isoelectric points, N--terminal amino acid , antigenicity, thermostability were tested. Results ADA both in Hippocampus tissue after reperfusion and in normal Hippocampus tissue have similar molecular weights, isoelectric points, N--terminal amino acid ,antigenicity. ADA＇s activity of normal Adenosine could he inhihited by p--chloramercuric benzoic acid ,and partially trstored by dithiothreital. But ADA＇ s activity of Hippocampus tissue after reperfusion could not he. On the other hand, ADA of Hippocampus tissue after reperfusion had much lower Km and stronger thermostahility compared with those in normal Hippocampus tissue. Conclusions The characteristic of ADA in Hippocampus tissue after reperfusion has been changed. The changes alter activity of ADA. Perhaps, those will determine Adenosine＇s effect in brain reperfusion injury.
出处
《神经疾病与精神卫生》
2006年第6期415-417,共3页
Journal of Neuroscience and Mental Health
关键词
腺苷脱氨酶
海马
缺血再灌注
Adenosine deaminase
Hippocampus tissue
Reperfusion
