肌苷对脑缺血再灌注后神经细胞凋亡和卡配因基因表达的影响  

EFFECTS OF INOSINE ON APOPTOSIS AND EXPRESSION OF CALPAIN mRNA IN NEURONS AFTER CEREBRAL ISCHEMIA AND REPERFUSION IN RATS

作　　者：张红[1] 王粤[2] 金丽英[1] 刘广义[1] 郭云良[1] 

机构地区：[1]青岛大学医学院脑血管病研究所,山东青岛266003 [2]青岛市人民医院神经内科

出　　处：《齐鲁医学杂志》2004年第1期14-16,19,共4页Medical Journal of Qilu

基　　金：山东省自然科学基金资助项目 (Y2 0 0 1C0 4)

摘　　要：①目的　研究肌苷对大鼠局灶性脑缺血再灌注后神经细胞凋亡和卡配因基因表达的影响 ,探讨肌苷的神经保护作用机制。②方法　取成年健康雌性SD大鼠 6 8只 ,随机分为治疗组和对照组 ,每组再随机分为缺血1.5h再灌注 2h、6h、12h、1d、2d、3d、7d、14d组 (n =4 ) ,另外 4只为假手术组。应用线栓法建立SD大鼠大脑中动脉阻塞 (MCAO)再灌注模型 ,腹腔注射肌苷注射液 (10 0mg/kg) ,原位末端标记 (TUNEL)和原位杂交技术分别观察大鼠脑缺血再灌注后神经细胞凋亡和卡配因mRNA表达。③结果　脑缺血再灌注后 2h皮质区与纹状体区即出现凋亡细胞 ,并逐渐增加 ,皮质区 1d达高峰 ,纹状体区 2d达高峰 ,之后逐渐减少 ,至 14d接近假手术组水平 ;经肌苷治疗后 ,皮质区和纹状体区凋亡神经细胞减少 ,其中再灌注 12h～ 7d较对照组有显著差异 (t=2 .5 2 5～ 8.987,P <0 .0 5 )。脑缺血再灌注后皮质区和纹状体区卡配因mRNA的表达于 2h逐渐增加 ,皮质区 12h、纹状体区1d达高峰 ,3d后逐渐降低 ,至 14d仍明显高于假手术组 ;肌苷治疗组再灌注 12h～ 14d ,其在皮质区和纹状体区表达较对照组显著降低 (t=4 .4 4 3～ 8.876 ,P <0 .0 5 )。④结论　脑缺血再灌注后肌苷使卡配因mRNA表达降低 ,神经细胞凋亡减少 ,提示肌苷具有抑制细胞凋亡和?ObjectiveTo investigate the effects of inosine on neuronal apoptosis and expression of calpain mRNA after focal cerebral ischemia and reperfusion(FCIR) in rats, and to explore the neuroprotective mechanisms of inosine.MethodsSixty-eight adult healthy female SD rats were randomly divided into three groups: treatment (n=32); control (n=32); and sham-operated (n=4). The rats in the treatment and control groups were subdivided into ischemia 90 min and reperfusion 2-, 6-, and 12-hour, and 1-, 2-, 3-, 7-, and 14-day subgroups, with four rats in each subgroup. A FCIR model was established by intraluminal middle cerebral artery occlusion(MCAO) with a nylon monofilament suture. Inosine(100 mg/kg) was injected intraperitoneally twice before and 12 hours after reperfusion. In situ hybridization was performed to examine the expression of calpain mRNA. Apoptosis was observed with terminal deoxynucleotidy1 transferase-mediated uridine 5′-triphosphate-biotin nick end-labeling(TUNEL) staining. ResultsCalpain mRNA expression was detected in the cortex and striatum of ischemic hemisphere as early as two hours after reperfusion and reached the peak at 12 hours and one day in the cortex and striatum respectively. Inosine treatment diminished the calpain mRNA expression. TUNEL-positive cells were observed two hours after reperfusion in the cortex and striatum and peaked at one and two days after reperfusion in the cortex and striatum, respectively. Furthermore, inosine decreased the number of TUNEL-positive cells at and 12 hours after reperfusion. ConclusionInosine attenuates ischemic neuronal apoptosis by inhibiting calpain mRNA expression, which suggests that inosine is effective to inhibit apoptosis and protect nerves. [

关 键 词：肌苷 脑缺血 再灌注损伤 神经细胞凋亡 卡配因 基因表达 

分 类 号：R743.3[医药卫生—神经病学与精神病学]