摘要
目的 探讨二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时NF-κB mRNA和fractalkine（FKN）mRNA表达的影响.方法 培养SD大鼠心肌微血管内皮细胞,以1×106个/ml的密度接种于96孔板（100μl/孔）或培养皿（2 ml/皿）,随机分为4组（n=24）.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2 h、复氧2 h.DZ组和DZ＋5-HD组在缺氧前2 h分别加入100 μmol/L二氮嗪、100 μmol/L二氮嗪＋100μmol/L 5-羟葵酸.于复氧2 h时测定细胞活力、细胞凋亡率、NF-κB mRNA和FKN mRNA表达水平.结果 与C组比较,H/R组细胞活力降低,细胞凋亡率升高,NF-κB mRNA和FKN mRNA表达上调（P＜0.01）;与H/R组比较,DZ组细胞活力升高,细胞凋亡率降低,NF-κB mRNA和FKN mRNA表达下调（P＜0.05）;5-羟葵酸可抑制二氮嗪预先给药导致的上述改变（P＜0.05）.结论 二氮嗪预先给药可下调NF-κB和FKN表达,从而减轻大鼠心肌微血管内皮细胞缺氧复氧损伤,其机制与激活线粒体ATP敏感性钾通道有关.
Objective To investigate the effects of diazoxide pretreatment on the expression of NF-κB mRNA and fractalkine （FKN） 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 （100μl/hole） or in 6 cm diameter dishes （2 ml/dish） with the density of 1 × 106/ml and randomly divided into4 groups （n = 24 each）： Ⅰ normal control group （group C）, Ⅱ H/R group, Ⅲ diazoxide pretreatment group （group DZ）, Ⅳ diazoxide pretreatment ＋ mitochondrial ATP-sensitive potassium channel blocker 5-hydroxydecanoate （5-HD） 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 100μmol/L were added to the cultured medium 2 h before hypoxia in group DZ and DZ ＋ 5-HD respectively. The cell vitality, apoptotic rate and expression of NF-κB mRNA and FKN mRNA were detected at end of reoxygenation. Results Compared with group C, the cell vitality was significantly decreased, apoptotic rate increased and the expression of NF-κB mRNA and FKN mRNA up-regulated in H/R group. Compared with group H/R, the cell vitality was significantly increased,apoptotic rate decreased and the expression of NF-κB mRNA and FKN mRNA down-regulated in group DZ. 5-HD could inhibit diazoxide pretreatment-induced changes mentioned above. Conclusion Diazoxide pretreatment can reduce H/R injury in rat myocardial microvascular endothelial cells through down-regulating the expression of NFκB and FKN, and the mechanism is related to activation of mitochondrial ATP-sensitive potassium channels.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2010年第11期1394-1396,共3页
Chinese Journal of Anesthesiology
