摘要
目的:了解脑红蛋白在正常大鼠脑中的分布以及缺氧缺血性脑损伤(hypoxic-ischemicdamage,HID)的表达,防治脑缺氧性损伤提供新思路。方法:①wistar大鼠,灌以40g/L多聚甲醛-磷酸缓冲液,取脑于40g/L多聚甲醛中4℃下固定,分子生物学常规方法液体细菌培养、提取纯化质粒、脑红蛋白质粒线性化、脑红蛋白cRNA探针标记,原位杂交组织化学方法检测脑红蛋白mRNA在正常大鼠脑中的分布。②Wistar大鼠,随机分为6组,其中5组制备脑缺氧缺血模型,分别在血流阻断1,5,15,30,60min断头取大脑,对照组为假手术组。设计脑红蛋白mRNA基因引物,提取脑组织总RNA,反转录合成单链cDNA,进行PCR反应后分析结果。Stata统计软件包oneway单因素方差分析进行数据处理。结果:①脑红蛋白mRNA阳性细胞在正常大鼠脑中定位于神经元中,分布广泛;扣带皮质,梨状皮质中的阳性细胞密集,阳性产物集中于细胞质;海马各区均有脑红蛋白mRNA阳性神经元分布;前脑核团中,杏仁中央核、杏仁内侧核、杏仁皮质前核、杏仁皮质后核、梨状内核均见脑红蛋白mRNA阳性神经元的分布,丘脑中脑红蛋白mRNA阳性神经元散在分布;小脑脑红蛋白mRNA阳性产物主要分布于蒲肯野细胞。②大鼠脑组织缺氧缺血时脑红蛋白mRNA表达呈现时间相的变化:缺血1min时。
AIM:To study the distribution of neuroglobin(NGB) gene in normal rat brain and the distribution of NGB gene in rats with hypoxic-ischemic damage (HID) so as to provide new way for the prevention and treatment of anoxic brain damage. METHODS:①After Wistar rats were perfused with 40 g/L paraformaldehyde phospho ric acid buffer,the brains were taken out and fixed in 40 g/L paraformaldehyde p hosphoric acid at 4℃.Liquid bacteria culture was performed with molecular biolo gy routine method,plasmid was purified and extracted,NGB plasmid linearity was m ade,NGBcRNA probe was used for labeling and distribution of NGBmRNA was detected in normal rat brain with in situ hybridization and histochemical method.②Wista r rats were randomized into 6 groups,and brain hypoxia-ischemia model was estab lished in the five groups,in which blood flow was blocked for 1,5,15,30 and 60 m inutes respectively.The rest one was used as control group(sham operation group) .Gene primer was designed,total RNA in brain tissue was extracted, mono-chain c DNA was synthesized through inverted-transcription,and results of PCR response was analyzed.The data were analyzed with Stata software by means of one-way mon o-agent analysis of variance. RESULTS:①Positive neurons of NGBmRNA were widely distributed in the normal ra t brains.The distribution of NGBmRNA positive cells was dense in cinguli cortex and pyriform cortex,and positive produce was concentrated in cytoplasm.NGBmRN po sitive cells were found in all the hippocampal areas. In forebrain nuclear mass, NGBmRN positive neurons were distributed in almond central nucleus,almond inside nucleus,almond cortex pronucleus, almond cortex postnucleus and piriform intran ucleus ether,while the NGBmRNA positive neurons in thalamus were scattered.Posit ive product of NGBmRNA in cerebellum was mainly distributed in Purkinje's cells. ②The expression of NGBmRNA was changed with time during hypoxia-ischemia as fo llows:At 1 minute of hypoxia-ischemia,the expression of NGBmRNA reached to the peak
出处
《中国临床康复》
CSCD
2004年第16期3060-3062,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30100049)~~
