颈内动脉注射质粒pLXSN-bcl-2对大鼠局灶性脑缺血后热休克蛋白70表达的影响  

Effect of Injecting Plasmid pLXSN-bcl-2 lntraventrically on the Expression of the HSP70 Protein after Transient Focal Cerebral Ischemia in Rats

作　　者：肇杰[1] 何志义[1] 陈玉 邓淑敏[1] 李宇[1] 

机构地区：[1]中国医科大学附属第一医院神经内科,沈阳110001 [2]辽宁省抚顺市矿业集团总医院神经内科,辽宁抚顺113008

出　　处：《中国医科大学学报》2008年第3期319-321,328,共4页Journal of China Medical University

基　　金：辽宁省自然科学基金资助项目(20032054)

摘　　要：目的观察bcl-2基因的过度表达在局灶性脑缺血中的脑保护作用,及能否引起热休克蛋白70(HSP70)表达的变化。方法参照改良的Longa栓线法构建大鼠局灶性脑缺血模型,缺血2h后将栓线拔出,经颈内动脉向大鼠脑内分别注人质粒pLXSN-bcl-2、空质粒pLXSN和生理盐水各260μl。存活的各组大鼠分别再灌注3,24,72h,断头取大脑。用免疫组织化学方法观察Bcl-2蛋白和HSP70蛋白表达情况。结果再灌注3h大鼠脑内即可见Bcl-2蛋白,HSP70表达,表达逐渐增加,二者均在再灌注24h达到高峰,再灌注72h表达降低,二者均以缺血区表达最强,但Bcl-2组两种蛋白的表达在各时间点均较对照组(空质粒组和生理盐水组)强,而对照组之间两种蛋白的表达在各个时间点均无差异。结论大鼠颈内动脉注射质粒pLXSN-bcl-2后,局灶性脑缺血大鼠脑组织中Bcl-2蛋白的广泛表达促进了HSP70基因的表达,保护了缺血脑组织。Objective To discuss the protective effect of the bcl-2 gene in the rat model of transient focal ischemia, and the relationship between the Bcl-2 protein and HSP70 family. Methods The rat model of transient focal cerebral ischemia was established with the modified Longa method and the suture was taken out 2 hours later. Then pLXSN plasmid （ 260 μl ）, pLXSN-bcl-2 plasmid （ 260 μl ）and normal saline were injected into the left internal carotid artery. All the living rots were decapitated at 3 h, 24 h, 72 h after repeffusion. Immunohistochemistry method was used to determine the expression of the Bcl-2 protein and the HSP70 protein, and the infarct area was delineated by triphenyhetrazolium chloride stain. Results The expression of Bcl-2 protein and HSP70 protein was seen 3 h after the repeffusion,and reached the peak 24 h, and then the expression decreased. The expression of Bcl-2 protein and HSPTO protein in mrs injected with pLXSN- bcl-2 was greater than that of the control groups at each time point,and much of the expression was found at the ＂penumbra field＂ and the hippocampus, caudate nucleus and thalamus, which were very sensitive to ischemic insult. Further more, the expression of Bcl-2 protein in rats injected with pLXSN-bcl-2 was extensive, while that in the control group was limited within the infarct focal site. Conclusion Injecting plasmid pLXSN-bcl-2 intraventrically into the internal carotid artery can increase the expression of the Bcl-2 protein in the ischemic rat brain. The overexpression of Bcl-2 protein can promote the expression of HSP70, and protect the cerebral tissue from ischemic insult.

关 键 词：脑缺血 基因治疗 质粒 BCL-2基因 热休克蛋白70 

分 类 号：R743.3[医药卫生—神经病学与精神病学]