大鼠肢体缺血-再灌注后脑CBS表达的变化及意义  

Changes of CBS expression in brain following ischemia and reperfusion of limbs

作　　者：王瑜玲[1] 马华民[2] 王冰[1] 张彦[1] 黄新莉[3] 

机构地区：[1]河北医科大学第三医院神经内科,河北石家庄050051 [2]河北医科大学第三医院药剂科,河北石家庄050051 [3]河北医科大学病理生理教研室,河北石家庄050017

出　　处：《中国病理生理杂志》2015年第2期213-218,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30800440);河北省自然科学基金资助项目(No.C2008001040;No.H2012206009;No.H2014206093188)

摘　　要：目的：观察肢体缺血-再灌注（IR）后脑组织内胱硫醚β-合酶（CBS）表达的变化及意义。方法：夹闭大鼠腹主动脉下段4 h、开放3-24 h复制肢体IR模型。采用逆转录多聚酶链反应检测脑内CBS mRNA表达的变化;Western blotting法检测脑内CBS蛋白的表达;采用全自动酶标仪测定脑组织中CBS活性和硫化氢含量;采用CBS抑制剂羟胺抑制CBS活性后,光镜观察脑组织的病理学变化。结果：肢体IR后脑组织CBS mRNA和蛋白表达水平明显高于假手术组,再灌12 h表达至峰值（均P〈0.01）,再灌注至24 h与假手术组比较无显著差异（均P〉0.05）。肢体IR后脑组织中CBS活性及硫化氢浓度的变化与CBS mRNA和蛋白表达的变化一致;与假手术组相比,单纯缺血组上述各指标均没有明显变化（均P〉0.05）;肢体IR后抑制CBS活性,IR组脑组织损伤加重。结论：肢体IR后,脑组织中CBS表达上调,所诱生的CBS对神经元具有保护效应。AIM： To detect the changes of cystathionine β-synthase（ CBS） expression in the brain following ischemia-reperfusion of hind limbs in the rats and to elucidate their significance. METHODS： Hind limb ischemia was induced by clamping infrarenal aorta with a microvascular clip and brain injury was made by following reperfusion. The brain tissue was obtained from the animals subjected to sham operation,4 h of ischemia without reperfusion and 3 h,6 h,12 h,24 h of reperfusion following 4 h of ischemia. The expression of CBS at mRNA and protein levels was measured at different time points by reverse transcription-polymerase chain reaction（ RT-PCR） and Western blotting. The CBS activity and hydrogen sulfide（ H2S） concentration in the brain were determined by a universal microplate reader. The observation of pathologic changes of the brain was made following the inhibition of CBS by hydroxylamine. RESULTS： The relative expression of CBS at mRNA and protein levels in IR group significantly increased compared with sham group. It reached a peak at 12h（ P〈0. 01）,and returned to baseline at 24 h（ P〈0. 05）. This time course correlated with increased CBS activity and H2 S concentration. No statistically significant difference in the above indexes between sham operation group and ischemia group was observed（ P〉0. 05）. Inhibition of CBS activity induced more severe brain injury in IR group. CONCLUSION：Up-regulation of CBS / H2 S pathway is involved in the protection for neurons during the ischemia-reperfusion of hind limbs.

关 键 词：脑 四肢 再灌注损伤 胱硫醚Β-合酶 硫化氢 

分 类 号：R363[医药卫生—病理学]