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目的 :了解缺血低氧刺激与大鼠心肌血管内皮生长因子 (VEGF)产生的关系、意义及其可能机制。方法 :1.建立Wistar大鼠急性心肌梗塞模型 ,将 2 8只大鼠随机分为 4组 ,每组 7只 :A组正常对照 ;B组急性心肌梗塞 1d ;C组急性心肌梗塞 3d ;D组急性心肌梗塞 7d。心肌冰冻切片 ,免疫组化探测VEGF。 2 .原代培养大鼠心肌细胞随机分组 :单纯缺氧组 ,A :缺氧培养 0h ;B :缺氧培养 6h ;C :缺氧培养 12h ;D :缺氧培养 2 4h ;蛋白激酶C(PKC)激动剂波佛酯 (PMA)组 ,A :加入PMA 0ng/mL ;B :加入PMA 10ng/mL ;C :加入PMA 10 0ng/mL ;D :加入PMA10 0 0ng/mL ;均缺氧培养 2 4h ;PKC抑制剂chelerythrine(CT)组 ,A :加入CT 0nmol/L ;B :加入CT 10nmol/L ;均缺氧培养 2 4h。免疫组化测定培养心肌细胞中血管内皮生长因子 (VEGF) ,经计算机扫描 ,图像分析定量。结果 :随缺血低氧时间延长 ,心肌产生VEGF增加 ,与对照组比较各组均有显著差异 ,缺血低氧时间与VEGF产生量呈正相关。不同剂量PMA与chelerythrine分别增强及减弱VEGF在缺氧心肌细胞中的表达。结论 :缺血低氧强烈刺激心肌产生血管内皮生长因子 ,缺血心肌通过分泌VEGF对自身具有重要的保护意义。其信号转导途径部分是通过PKC通路实现的。
Objective: To investigate the relationship between the expression of vascular endothelial growth factor (VEGF) and ischemic hypoxia in rats and its possible mechanism. Acute myocardial infarction model was established in Wistar rats, 28 rats were randomly divided into 4 groups with 7 rats in each group: normal control group A, acute myocardial infarction group 1d, acute myocardial infarction group C 3d, Group Acute myocardial infarction 7d. Myocardial frozen section, immunohistochemical detection of VEGF. The primary cultured rat cardiomyocytes were randomly divided into three groups: hypoxia group, A: hypoxia for 0h, B: hypoxia for 6h, C: hypoxia for 12h, D: hypoxia for 24h, protein kinase C PKC agonist wave-PMA group, A: adding PMA 0ng / mL; B: adding PMA 10ng / mL; C: adding PMA 10ng / mL; D: adding PMA10 0ng / mL; 2 4h; PKC inhibitor chelerythrine (CT) group, A: adding CT 0nmol / L; B: adding CT 10nmol / L; hypoxic culture 24h. The expression of vascular endothelial growth factor (VEGF) in cardiomyocytes was detected by immunohistochemistry and quantified by computerized scanning and image analysis. Results: With the prolongation of ischemia and hypoxia, myocardial VEGF increased, compared with the control group, there was a significant difference between each group. There was a positive correlation between the time of hypoxia and the amount of VEGF production. Different doses of PMA and chelerythrine enhance and weaken the expression of VEGF in hypoxic cardiomyocytes, respectively. CONCLUSION: Ischemia and hypoxia strongly stimulate the production of vascular endothelial growth factor in myocardium. It is important that ischemic myocardium have its own protective significance by secreting VEGF. Part of its signal transduction pathway is through the PKC pathway.