【作者】 余福林；

【导师】 王晨霞；

【作者基本信息】 延安大学 ， 内科学， 2011， 硕士

【摘要】 研究目的:1.建立离体心脏缺血/再灌注损伤模型,监测大鼠的心功能变化并测定心肌生化指标,验证离体缺血/再灌注损伤模型建立成功。2.成功建立缺血/再灌注损伤模型后,探讨缺血/再灌注损伤后心肌细胞线粒体功能及其能量代谢变化。研究方法:1. SD雄性大鼠脱颈椎处死,迅速取出心脏后投入到4℃的K-H液中,行主动脉插管,丝线结扎固定,挂靠在Langendorff灌流装置,连接压力换能器传感器,开始灌流;RM-6280多道生理记录和分析系统记录分析心功能变化;收集灌流后液体,测定心肌生化指标。2.将实验动物分为正常对照组、单纯缺血组和缺血再灌注组(n=8)。各组灌流结束后,取大鼠左室全层心肌作为标本提取目的蛋白AMPK和Cyt– C。Western blot法测定心肌细胞AMPKα和Cyt - C的蛋白表达。研究结果:1.离体缺血/再灌注损伤模型结果:与对照组相比,实验组的LVSP(P <0.05)、+dp/dtmax(P <0.05)与-dp/dtmax(P <0.05)降低;LVEDP(P <0.05)升高,两组相比差异显著;而两组HR差异不显著(P >0.05)。I/R组心肌生化指标CK、LDH、CK-MB与实验组比较明显升高(P <0.05)。提示大鼠离体心脏缺血再灌注模型建立成功。2.缺血再灌注组AMPKα的蛋白表达显著降低(P<0.05)。而细胞色素C的蛋白表达均显著升高(P<0.05)。研究结论:1.成功建立离体缺血/再灌注损伤模型。2.缺血/再灌注损伤后心肌细胞能量蛋白AMPKα1的蛋白表达显著降低,而细胞色素C的蛋白表达均显著升高。更多还原

【Abstract】 Objective: 1. to establish in vitro ischemia / reperfusion injury model in rats to monitor changes in cardiac function and biochemical indicators of myocardial;Verify the in vitro ischemia / reperfusion injury model was established successfully. 2. Successfully established ischemia / reperfusion injury model; then test the mitochondrial function and energy metabolism.Methods: 1. SD rats were killed off the spine, removed the heart quickly into 4℃KH solution, aortic cannulation, silk ligation fixed, anchored in the Langendorff perfusion apparatus, connecting the pressure transducer sensor, start perfusion; RM-6280 multi-channel recording and analysis system recorded the physiological analysis of cardiac function; collecting fluid after perfusion and testing the myocardial biochemical indicators. 2. The experimental animals were divided into normal control group, ischemia-reperfusion group and ischemia group (n = 8). After perfusion in each group, taking the left ventricular myocardial to extract the protein of AMPK and Cyt - C. using Western blot to determine the protein of AMPKαand Cyt - C.Results: 1.The results In vitro ischemia / reperfusion injury model: Compared with the control group, LVSP (P <0.05), + dp / dtmax (P <0.05) and -dp/dtmax (P <0.05 ) decreased in experimental group; LVEDP (P <0.05) increased in experimental group with a significant difference; and HR groups was not significant (P > 0.05). Compared with the experimental group, I / R myocardial biochemical indicators CK, LDH, CK-MB was significantly higher (P <0.05). We can say the in vitro ischemia-reperfusion model is established successfully. 2. The expression of AMPKαprotein was significantly decreased (P <0.05) in Ischemia-reperfusion group. The expression of cytochrome C protein were significantly higher (P <0.05). Conclusions: 1. the in vitro ischemia / reperfusion injury model was successfully established. 2. The expression of AMPKαprotein was significantly decreased (P <0.05) in Ischemia-reperfusion group, but the expression of cytochrome C protein was significantly increased.更多还原