大鼠脊髓缺血再灌注损伤后CaMKII的表达及作用研究  

The expression and role of Ca MKII after spinal cord ischemia-reperfusion injury in rats

作　　者：仝淞铭 曹阳[1] 于德水[1] 魏子健[1] 刘超[1] 武林[1] 王建[1] 

机构地区：[1]辽宁医学院附属第一医院骨科,辽宁锦州121001

出　　处：《中国矫形外科杂志》2015年第12期1117-1122,共6页Orthopedic Journal of China

基　　金：国家自然科学基金资助基金(编号:81471853);辽宁省博士科研启动基金(编号:20121096);辽宁省高等学校优秀人才支持计划项目(编号:LR2013091);辽宁医学院博士及出国进修1年以上教师科研启动基金项目(编号:Y2012B011);辽宁医学院校长基金-奥鸿博泽研究生科技创新基金(编号:AH2014019)

摘　　要：[目的]探讨钙调蛋白依赖性蛋白激酶2(Ca MKII)在大鼠脊髓缺血再灌注损伤中作用。[方法]雄性SD大鼠56只,随机分为假手术组(n=8)、实验组(n=48),假手术组暴露肾动脉下腹主动脉,但不阻断,实验组采用Naslund腹主动脉阻断法制作脊髓缺血再灌注损伤模型,于再灌注后0、1、3、7、14、28 d取脊髓标本,HE染色观察脊髓病理变化,免疫组化和Westewrn blot检测Ca MKII的表达情况,TUNEL染色检测神经细胞凋亡。[结果]实验组脊髓组织出现明显病理学改变。Western blot在56KD分子量处可见Ca MKII蛋白特异性条带,假手术组Ca MKII表达较少,实验组0 d即出现Ca MKII表达增多,且随着时间延长表达量逐渐增多,于3 d达高峰后逐渐降低,至14 d时仍高于假手术组(P<0.01)。免疫组化结果显示,Ca MKII蛋白主要表达于脊髓神经元胞浆,Ca MKII阳性细胞数的变化趋势与免疫组化结果相似。TUNEL染色示假手术组有少量凋亡细胞散在分布,实验组术后0 d开始出现凋亡细胞增多,3 d达高峰,后逐渐减少,28 d时仍高于假手术组(P<0.05)。[结论]脊髓缺血再灌注损伤后Ca MKII表达增加,同时细胞凋亡数增多,Ca MKII可能通过调节细胞凋亡来影响脊髓缺血再灌注损伤。[Objective]To explore the role of Ca MKII in spinal cord ischemia- reperfusion injury rats. [Method]Fifty-six healthy adult SD rats,were randomly divided into sham group（ n = 8） and injuried group（ n = 48）,sham group rats were only subjected to exposure the abdominal aorta below renal artery but without occlusion,injuried group rats were created by using Naslund abdominal aorta occlusion,injuried group were harvested at 0 day,1 day,3 days,7 days,14 days,28 days after surgery.HE staining was used to observe the pathological changes of spinal cord,the expression of Ca MKII was evaluated by immunohistochemistry and Western blot. TUNEL staining was used to detect apoptosis cells. [Result]The injuried group showed the pathological changes obviously. Western blot specific bands of Ca MKII visible at a molecular weight of 56 k D,Ca MKII expressed less in sham group,0 day after reperfusion,the expression of Ca MKII began to increase,the expression of Ca MKII gradually increased as time went along,reached a peak at 3 days and then decreased,until 14 days,still higher than in sham group（ P〈0. 01）. Immunohistochemical results suggested that Ca MKII had expressed in the cytoplasm of neurons,the trends of Ca MKII positive cells were similar with the western blot results. TUNEL positive cells distributed in sham group,0 day after reperfusion,the apoptotic cells began to increase,reached a peak at 3 days,reduced gradually,and at 28 days were still higher than in sham group（ P〈0.05）. [Conclusion]The expression of Ca MKII was significantly increased after spinal cord ischemia- reperfusion injury,the apoptotic cells also increased. Ca MKII may mediat cell apoptosis to influence spinal cord ischemia- reperfusion injury.

关 键 词：脊髓损伤 缺血再灌注 凋亡 Ca MKII 

分 类 号：R651.2[医药卫生—外科学]