针刺预处理全脑缺血大鼠海马CA1区细胞凋亡和热休克蛋白70 mRNA的表达  

Effect of acupuncture precondition on apoptosis and heat shock protein 70 mRNA expression in hippocamal CA1 of rats with global cerebral ischemia

作　　者：唐伟[1] 孙忠人[2] 张力[2] 王威[3] 

机构地区：[1]大连大学附属医院神经内科,辽宁省大连市116021 [2]黑龙江中医药大学附属第二医院神经内科,黑龙江省哈尔滨市150001 [3]大连理工大学神经信息研究所,辽宁省大连市116000

出　　处：《中国临床康复》2005年第37期90-92,共3页Chinese Journal of Clinical Rehabilitation

基　　金：辽宁省教育厅高等学校科学研究项目(20040050)~~

摘　　要：目的:探讨针刺预处理对全脑缺血大鼠海马CA1区细胞凋亡及热休克蛋白70mRNA表达的影响,并与缺血预处理的效果进行比较。方法:实验于2003-10在黑龙江中医药大学神经解剖实验室进行。取120只Wistar大鼠随机分为5组,每组24只:①正常对照组:不干预。②假手术组:暴露4条血管,不造模。③脑缺血组:四动脉阻断法全脑缺血10min制作大鼠全脑缺血模型。④脑缺血预处理组:预全脑缺血3min,再灌注24h后再次全脑缺血10min。⑤针刺预处理组:术前7d给予针刺,双侧足三里、曲池(,双侧连接全能脉冲电疗仪,频率为1Hz,电压为2V,30min/次,针刺百会30min/次,1次/d,7d后全脑缺血10min。每组分别于再灌注12,24,48和72h麻醉状态下取材,采用原位缺口末端标记法检测大鼠脑海马CA1区凋亡细胞数,免疫组织化学法检测大鼠脑海马热休克蛋白70阳性细胞数,原位分子杂交技术检测大鼠脑海马热休克蛋白70mRNA表达。结果:经补充后120只大鼠进入结果分析。①脑海马CA1区凋亡细胞数:正常对照组和假手术组无;脑缺血组再灌注12h即可见,且随时间延长逐渐增多,72h仍具有较高水平[(55.87±10.68)个/mm2],而脑缺血预处理组和针刺预处理组各时间点均显著低于脑缺血组(P<0.05),但两组间无差异(P>0.05)。②脑海马热休克蛋白70阳性细胞数:正常对照组和假手术组无;脑缺血组少量表达,再灌注24h为高峰[(20.84±5.93)个/mm2],而脑缺血预处理组和针刺预处理组各时间点均显著高于脑缺血组(P<0.05),但两组间无差异(P>0.05)。③脑海马热休克蛋白70mRNA阳性细胞数:正常对照组和假手术组无;脑缺血组少量表达,再灌注48h为高峰[(19.44±5.55)个/mm2],而脑缺血预处理组和针刺预处理组各时间点均显著高于脑缺血组(P<0.05),但两组间无差异(P>0.05)。结论:针刺预处理的脑保护机制可能与抑制细胞凋亡及上调热休克蛋白70mRNA、热休克蛋白70表达水�AIM： To explore the effect of acupuncture precondition on apoptosis and heat shock protein 70 （HSP70） mRNA expression in hippocamal CA1 of cerebral ischemic rats in comparison with ischemic precondition. METHODS： The experiment proceeded in the Neuroanatomy Laboratory of Heilongjiang University of Traditional Chinese Medicine in October 2003. 120 Wistar rats were randomized into 5 groups with 24 in each group： ① normal control group： no intervention; ②sham-operation group： only to expose 4 blood vessels; ③ischemic group： to block 4 blood vessels for 10 minutes; ④ ischemic precondition group： to block 4 blood vessels for 3 minutes firstly and then 10 minutes after 24 hours; ⑤ acupuncture precondition group： to electroacupuncture at Zusanli and Quchi in 2 V and 1 Hz once for 30 minutes and acupuncture at Baihui for 30 minutes every day, and at last global cerebral ischemia for 10 minutes after 7 days. All the rats were killed under anesthesia at hours 12, 24, 48 and 72 after reperfusion to measure the amount of apoptotic cells with TUNEL method, the number of HSP70-positive cells with immunohistochemical method and the expression of HSP70 mRNA with in suit molecular hybridization technique. RESULTS： After supplement, 120 rats were involved in the result analysis. ①Apoptotic cells in hippocampal CA1： There was no difference between the control and sham operation group. Apoptotic cells were found in the ischemic group at 12 hours after reperfusion, then increased gradually and still maintained a higher level at 72 hours （55.87±10.68）.The amount of apoptotic cells in the ischemic precondition group and the acupuncture precondition group was markedly lower than that in the ischemic group at any time points（P〈0.05）, but there was no difference between the two former groups（P〉0.05）. ②HSP70-positive cells in hippocampus： No positive cells were found in the control group and sham operation group. A few were found in the ischemic group, and reached the peak at 24 hour

关 键 词：脑缺血/针灸疗法 细胞凋亡 热休克蛋白70 缺血预处理 

分 类 号：R743[医药卫生—神经病学与精神病学]