CAJ | 학술논문

背景:细胞凋亡与细胞色素C(cytochrome C,Cyt C)、卡配因、Bcl-2家族和半胱天冬酶家族等基因表达有密切关系.脑缺血再灌注过程中,下调卡配因的表达,抑制Cyt C的释放可减少细胞凋亡.目的:研究大鼠局灶性脑缺血再灌注后神经细胞凋亡及其与Cyt C和卡配因基因表达的关系.设计:随机对照的实验研究.地点和材料:实验地点:青岛大学医学院脑血管病研究所和山东省脑血管病防治重点实验室.动物:成年健康雌性SD大鼠36只,体质量230～270 g,清洁级,由中国科学院上海实验动物中心提供.方法:全部实验均由所有作者完成.具体方法:应用线栓法建立SD大鼠大脑中动脉阻塞(middle cerebral artery occlusion,MCAO)再灌注模型,应用TUNEL法观察脑缺血再灌注后神经细胞凋亡,原位杂交技术检测Cyt C mRNA和卡配因mRNA的表达.主要观察指标:凋亡细胞数;Cyt C和卡配因的阳性细胞数.结果:脑缺血再灌注后2 h脑组织即出现凋亡细胞,在皮质区和纹状体区分别于1 d和2 d达高峰,此后逐渐减少.脑组织Cyt C mRNA和卡配因mRNA的表达自缺血再灌注后2 h后逐渐增高,皮质区12 h达高峰[分别为(122.50±6.69)和(138.50±6.25)个/视野],纹状体区1 d达高峰[分别为(119.25±5.12)和(105.00±4.58)个/视野],以后逐渐下降.Cyt C mRNA和卡配因mRNA的表达与细胞凋亡的区域基本一致.结论:脑组织皮质区神经细胞较纹状体区对缺血性损伤更为敏感,Cyt C和卡配因基因表达在诱导细胞凋亡中具有重要作用.
배경:세포조망여세포색소C(cytochrome C,Cyt C)、잡배인、Bcl-2가족화반광천동매가족등기인표체유밀절관계.뇌결혈재관주과정중,하조잡배인적표체,억제Cyt C적석방가감소세포조망.목적:연구대서국조성뇌결혈재관주후신경세포조망급기여Cyt C화잡배인기인표체적관계.설계:수궤대조적실험연구.지점화재료:실험지점:청도대학의학원뇌혈관병연구소화산동성뇌혈관병방치중점실험실.동물:성년건강자성SD대서36지,체질량230～270 g,청길급,유중국과학원상해실험동물중심제공.방법:전부실험균유소유작자완성.구체방법:응용선전법건립SD대서대뇌중동맥조새(middle cerebral artery occlusion,MCAO)재관주모형,응용TUNEL법관찰뇌결혈재관주후신경세포조망,원위잡교기술검측Cyt C mRNA화잡배인mRNA적표체.주요관찰지표:조망세포수;Cyt C화잡배인적양성세포수.결과:뇌결혈재관주후2 h뇌조직즉출현조망세포,재피질구화문상체구분별우1 d화2 d체고봉,차후축점감소.뇌조직Cyt C mRNA화잡배인mRNA적표체자결혈재관주후2 h후축점증고,피질구12 h체고봉[분별위(122.50±6.69)화(138.50±6.25)개/시야],문상체구1 d체고봉[분별위(119.25±5.12)화(105.00±4.58)개/시야],이후축점하강.Cyt C mRNA화잡배인mRNA적표체여세포조망적구역기본일치.결론:뇌조직피질구신경세포교문상체구대결혈성손상경위민감,Cyt C화잡배인기인표체재유도세포조망중구유중요작용.
BACKGROUND: Programmed cell death has a close relationship with gene expression, such as cytochrome C(Cyt C), Calpain, Bcl-2 family and caspases. Down-regulation of calpaine gene expression during ischemia-reperfusional(IR) injury may inhibit Cyt C release, which results in attenuated cell apoptosisOBJECTIVE: To investigate the correlation of neuronal apoptosis with gene expressions of Cyt C and calpain in rats after focal cerebral IR injury.DESIGN: Randomized experimental case-control study.SETTING and MATERIALS: The study was carried out at the Cerebrovascular Disease Research Institute of Qingdao Medical College and Cerebrovascular Disease Prevention Laboratory of Shandong Province. Thirty-six healthy clean adult female SD rats, weighing 230 - 280 g,were provided by Shanghai Experimental Animal Center of Chinese Academy of Science.METHODS: Reperfusion animal models of middle cerebral artery occlusion (MCAO) were established on SD rat by thread-bolt method. TUNNEL method was used to observe neuronal apoptosis after IR and in situ hybridization technique was applied to detect the expressions of Cyt C mRNA and Calpain mRNA.MAIN OUTCOME MEASURES: Numbers of apoptosis neurons and Cyt C and Calpain-positive cells.RESULTS: Neuron apoptosis occured 2 hours after cerebral IR, increased to the peak 1 day and 2 days after IR in cortex and striatum, respectively, and then gradually decreased. Cerebral Cyt C mRNA and Calpain mRNA expressions increased 2 hours after IR, and to the peak at 12 hours in cortex [ (122.50 ± 6. 69) and( 138. 50 ± 6.25)-eye field, respectively], and 1 day in striatum[ ( 119.25 ± 5.12) and( 105.00 ± 4. 58)-eye field, respectively ],and then decreased gradually. The spacial expression of Cyt C mRNA and calpain mRNA were consistent with the region of neuronal apoptosis.CONCLUSION: Neurons in cortex were more vulnerable to ischemic injury than those in striatum. Cyt C and calpain gene expressions play a key role in inducing cell apoptosis.