二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时PI3K mRNA和AktmRNA表达的影响  

Effects of diazoxide pretreatment on expression of phosphatidylinositol 3-kinase mRNA and protein serine/ threonine kinase mRNA in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation

作　　者：曹苏[1] 陈秋萍[1] 康焕菊[1] 沈施仁[1] 

机构地区：[1]南通大学附属医院麻醉科,226001

出　　处：《中华麻醉学杂志》2011年第7期871-873,共3页Chinese Journal of Anesthesiology

基　　金：江苏省卫生厅科技项目（z201018）;南通大学自然科学项目（102048）

摘　　要：目的评价二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时磷脂酰肌醇．3激酶（P13K）mRNA和蛋白质丝氨酸苏氨酸激酶（Akt）mRNA表达的影响。方法培养sD大鼠心肌微血管内皮细胞，以1×10^6个／ml密度接种于96孔培养板（100μl／孔）或培养皿（2ml／血），采用随机数字表法，将其随机分为4组（n=25）。正常对照组（C组）不作任何处理；缺氧复氧组（H／R组）、二氮嗪预先给药组（DZ组）、二氮嗪预先给药＋线粒体ATP敏感性钾通道阻断剂5－羟葵酸组（DZ＋5－HD组）均进行缺氧2h，复氧2h。DZ组和DZ＋5-HD组在缺氧前2h分别加入100μmol／L二氮嗪、100gmol／L二氮嗪＋100μmol／L5-羟葵酸。于复氧2h时测定细胞活力、细胞凋亡率、P13KmRNA和AktmRNA表达。结果与c组比较，H／R组细胞活力降低，细胞凋亡率升高，P13KmRNA和AktmRNA表达上调（P〈0．05或0．01）；与H／R组比较，DZ组细胞活力升高，细胞凋亡率降低，P13KmRNA和AktmRNA表达上调（P〈0．05或0．01）；5-羟葵酸可逆转二氮嗪预先给药导致的上述改变（P〈0．05或0．01）。结论二氮嗪预先给药减轻大鼠心肌微血管内皮细胞缺氧复氧损伤的机制与激活线粒体ATP敏感性钾通道，促进P13K和Akt基因的转录有关。Objective To investigate the effects of diazoxide pretreatment on expression of phosphatidylinositoi 3-kinase（PI3K） mRNA and protein serine/threonine kinase（Akt） mRNA in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation （H/R）. Methods The SD rat myocardial microvascular endothelial cells were cultured. The cells were seeded in 96-well plates （100 μl/hole） or in 6 cm diameter dishes （2 ml/dish） with the density of 1 × 10^6/ml and randomly divided into 4 groups （ n = 25 each） ： normal control group （group C）, H/R group, diazoxide pretreatment group （group DZ） and diazoxide pretreatment ＋ 5-hydroxydecanoate （5-HD, a mitochondrial ATP-sensitive potassium channel blocker） group （group DZ ＋ 5-HD） . The cells were exposed to 2 h hypoxia followed by 2 h reoxygenation. Diazoxide 100 μmol/L and diazoxide 100 μmol/L ＋ 5-HD 100 μmol/L were added to the culture medium 2 h before hypoxia in groups DZ and DZ ＋ 5-HD respectively. The cell viability, apoptotic rate and expression of PI3K mRNA and Akt mRNA were detected at the end of reoxygenation. Results Compared with group C, the cell viability was significantly decreased, while the apoptotic rate increased and expression of PI3 K mRNA and Akt mRNA up-regulated in group H/R （P 〈 0.05 or 0.01 ）. Compared with group H/R, the cell viability was significantly increased, while the apoptotic rate decreased and expression of PI3K mRNA and Akt mRNA up-regulated in group DZ （P 〈 0.05 or 0.01 ）. 5-HD could inhibit diazoxide pretreatment-induced changes mentioned above （ P 〈 0.05 or 0.01 ）. Conclusion Diazoxide pretreatment can reduce H/R injury by promoting PI3K gene and Akt gene transcription through activation of mitochondrial ATP-sensitive potassium channels in rat myocardial microvascular endothelial cells.

关 键 词：二氮嗪 心脏 内皮 血管 氧 细胞低氧 1-磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 RNA 信使 

分 类 号：R965[医药卫生—药理学]