CAJ | 학술논문

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细胞凋亡的影响.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孔培养板(100 μl/孔)或培养皿(2 ml/皿),采用随机数字表法,将其随机分为4组(n=12),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)和二氮嗪预先给药+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二氮嗪和100 μmol/L二氮嗪+100 μmol/L线粒体ATP敏感性钾通道阻断剂5-羟葵酸.于复氧2 h时测定细胞活力和凋亡率.结果与C组比较,H/R组细胞活力降低,细胞凋亡率升高(P＜0.01);与H/R组比较,DZ组细胞活力升高,细胞凋亡率降低(P＜0.05或0.01);5-羟葵酸可抑制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可抑制大鼠心肌微血管内皮细胞凋亡,从而减轻缺氧复氧损伤,其机制与激活线粒体ATP敏感性钾通道有关.
목적 탐토이담진예선급약대대서심기미혈관내피세포결양복양시세포조망적영향.방법배양SD대서심기미혈관내피세포,이1×106개/ml적밀도접충우96공배양판(100 μl/공)혹배양명(2 ml/명),채용수궤수자표법,장기수궤분위4조(n=12),정상대조조(C조)불작임하처리,결양복양조(H/R조)、이담진예선급약조(DZ조)화이담진예선급약+5-간규산조(DZ+5-HD조)균진행결양2 h복양2 h,DZ조화DZ+5-HD조재결양전2 h분별가입100 μmol/L이담진화100 μmol/L이담진+100 μmol/L선립체ATP민감성갑통도조단제5-간규산.우복양2 h시측정세포활력화조망솔.결과 여C조비교,H/R조세포활력강저,세포조망솔승고(P＜0.01);여H/R조비교,DZ조세포활력승고,세포조망솔강저(P＜0.05혹0.01);5-간규산가억제이담진예선급약도치적상술개변(P＜0.05혹0.01).결론 이담진예선급약가억제대서심기미혈관내피세포조망,종이감경결양복양손상,기궤제여격활선립체ATP민감성갑통도유관.
Objective To investigate the effects of diazoxide pretreatment on apoptosis 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 1×106/ml and randomly divided into 4 groups ( n = 12 each) : normal con trol 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 and apoptotic rate were detected at the end of reoxygenation. Results Compared with group C, the cell viability was significantly decreased, while the apoptotic rate increased in group H/R ( P ＜ 0.01) . Compared with group H/R, the cell viability was significantly increased, while the apoptotic rate decreased 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 through inhibiting apoptosis in rat myocardial microvascular endothelial cells, and the mechanism is related to the activation of mitochondrial ATP-sensitive potassium channels.