Effects of ganglioside on cell apoptosis of rats with acute cerebral ischemia-reperfusion injury / 中国组织工程研究

ABSTRACT

BACKGROUND: Cell apoptosis and expression of related apoptotic gene are found in rats with acute cerebral ischemia-reperfusion injury.OBJECTIVE: To study the effects of ganglioside on cell apoptosis in rats with acute cerebral ischemia-reperfusion injury.DESIGN: Randomized control animal experiment.SETTING: Department of Neurology, China-Japan Friendship Hospital of Jilin University.MATERIALS The experiment was conducted in the Animal Laboratory of China-Japan Friendship Hospital of Jilin University at April 2002.Forty-eight healthy male Wistar rats aged 3-4 months with the body mass of (220±50) g were selected and randomly divided into ischemia-reperfusion group and ischemia-reperfusion + administration group (intraperitoneal injection of ganglioside GM-1 at 30 minutes before ischemia) with 24 rats in each group, and each group was subdivided into three groups according to the reperfusion time 3-hour, 6-hour and 24-hour with 8 rats in each time-point.METHODS: ①Rat models of cerebral ischemia-reperfusion were established. ②Diphenylamine method was adopted to detect changes of DNA splitting rate in brain tissues at 3 hours,6 hours and 24 hours after cerebral ischemia.100 mg of cerebral cortex was made into 10% homogenate by adding into 0.9 mL of splitting fluid,which was then put in the centrifuge tube for repeated freezing and melting. The supernatant and deposit were collected.DNA splitting rate = supernatant absorption/(supernatant absorption + deposit absorption). ③Immunohistochemical method was used to study the expression of protein kinase Cδ (PKCδ) as well as changes of ganglioside after administration. MAIN OUTCOME MEASURES: Changes of DNA splitting rate in cerebral cortex of rats as well as intensity of PKCδ expression.RESULTS: One rat in the normal saline control group died for exceeding anesthetization at the 6th hour of reperfusion, and 2 rats died at the 24th hour of reperfusion, which were supplemented respectively. With the time of reperfusion increasing, changes of DNA splitting rate significantly increased, which peaked at the 24th hour. The expression of PKCδ peaked at the 6th hour of reperfusion and gradually decreased. The DNA splitting rate and PKCδ expression were remarkably decreased at corresponding time-points in the ganglioside group.CONCLUSION: Ganglioside can inhibit cell apoptosis and reduce PKCδ expression after cerebral ischemia-reperfusion to protect cerebral ischemia-reperfusion injury.