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用雄性Wistar大鼠36只,形成三动脉结扎全脑缺血模型,用改良的异硫氰酸胍一步法从脑组织中提取总RNA,与a-~(32)P标记的c-fos cDNA(4.8kb)探针进行斑点杂交,观察大鼠脑缺血过程中c-fosmRNA表达的变化,动物分别在全脑缺血30min后,再灌0min,15min、30min、60min,90min,120min,150min、180min后断头处死,每次处死3只,假手术对照组(3只)除不夹闭血管外,处理同缺血组。结果显示:假手术对照组c-fos mRNA的斑点灰度为1.07±0.25,再灌15min呈上升趋势
Thirty-six male Wistar rats were subjected to three-artery occlusion of global cerebral ischemia, and total RNA was extracted from the brain tissue by one-step guanidinium isothiocyanate method. The total RNA was extracted from brain tissue and incubated with a- (32) P-labeled c-fos cDNA (4.8kb) were used to detect the expression of c-fos mRNA in rat brain after cerebral ischemia. The animals were subjected to ischemia for 30 min, reperfusion for 0 min, 15 min, 30 min, 60 min, 90 min, 120 min, 150 min After 180 minutes, the rats were decapitated and sacrificed 3 times each. The sham operation control group (3) was treated with the same ischemia group except for not clamping the blood vessel. The results showed that the gray level of c-fos mRNA in control group was 1.07 ± 0.25