Wu Haidong, Wang Peng, Li Yi, Wu Manhui, Lin Jiali, Huang Zitong
Department of Emergency Medicine, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China; Institute of Cardiopulmonary Cerebral Resuscitation, Sun Yat-sen University, Guangzhou, China.
Biomed Res Int. 2016;2016:1253842. doi: 10.1155/2016/1253842. Epub 2016 Aug 28.
Objective. We investigated whether and how diazoxide can attenuate brain injury after cardiopulmonary resuscitation (CPR) by selective opening of mitochondrial ATP-sensitive potassium (mitoKATP) channels. Methods. Adult male Sprague-Dawley rats with induced cerebral ischemia (n = 10 per group) received an intraperitoneal injection of 0.1% dimethyl sulfoxide (1 mL; vehicle group), diazoxide (10 mg/kg; DZ group), or diazoxide (10 mg/kg) plus 5-hydroxydecanoate (5 mg/kg; DZ + 5-HD group) 30 min after CPR. The control group (sham group, n = 5) underwent sham operation, without cardiac arrest. Mitochondrial respiratory control rate (RCR) was determined. Brain cell apoptosis was assessed using TUNEL staining. Expression of Bcl-2, Bax, and protein kinase C epsilon (PKCε) in the cerebral cortex was determined by Western blotting and immunohistochemistry. Results. The neurological deficit scores (NDS) in the vehicle group decreased significantly at 24 h and 48 h after CPR. Diazoxide significantly improved NDS and mitochondrial RCR after CPR at both time points; 5-HD cotreatment abolished these effects. Diazoxide decreased TUNEL-positive cells following CPR, upregulated Bcl-2 and PKCε, downregulated Bax, and increased the Bcl-2/Bax ratio; 5-HD cotreatment reversed these effects. Conclusions. Diazoxide attenuates postresuscitation brain injury, protects mitochondrial function, inhibits brain cell apoptosis, and activates the PKC pathway by opening mitoKATP channels.
目的。我们研究了二氮嗪是否以及如何通过选择性开放线粒体ATP敏感性钾通道(mitoKATP)来减轻心肺复苏(CPR)后的脑损伤。方法。诱导脑缺血的成年雄性Sprague-Dawley大鼠(每组n = 10)在心肺复苏后30分钟接受腹腔注射0.1%二甲基亚砜(1毫升;载体组)、二氮嗪(10毫克/千克;DZ组)或二氮嗪(10毫克/千克)加5-羟基癸酸(5毫克/千克;DZ + 5-HD组)。对照组(假手术组,n = 5)接受假手术,无心脏骤停。测定线粒体呼吸控制率(RCR)。使用TUNEL染色评估脑细胞凋亡。通过蛋白质印迹法和免疫组织化学法测定大脑皮质中Bcl-2、Bax和蛋白激酶Cε(PKCε)的表达。结果。载体组的神经功能缺损评分(NDS)在心肺复苏后24小时和48小时显著降低。二氮嗪在两个时间点均显著改善了心肺复苏后的NDS和线粒体RCR;5-HD联合治疗消除了这些作用。二氮嗪降低了心肺复苏后的TUNEL阳性细胞,上调了Bcl-2和PKCε,下调了Bax,并增加了Bcl-2/Bax比值;5-HD联合治疗逆转了这些作用。结论。二氮嗪减轻复苏后脑损伤,保护线粒体功能,抑制脑细胞凋亡,并通过开放mitoKATP通道激活PKC途径。