盐酸曲马多预处理对去甲肾上腺素诱导大鼠离体心肌细胞核转录因子-κB活化的影响  

Effects of tramadol hydrochloride preconditioning on the activation of nuclear factor-κB induced by norepinephrine in cultured neonatal rat cardiomyocytes

作　　者：吕志敢[1] 郭政[2] 

机构地区：[1]山西医学科学院、山西大医院麻醉科,太原030032 [2]山西医科大学第二医院麻醉科,太原030001

出　　处：《中华危重病急救医学》2013年第11期646-649,共4页Chinese Critical Care Medicine

基　　金：国家自然科学基金（30471656）

摘　　要：目的 观察盐酸曲马多预处理对去甲肾上腺素（NE）诱导大鼠离体心肌细胞核转录因子-κB（NF-κB）活化的影响.方法 采用1～3日龄新生SD大鼠建立体外培养心肌细胞实验模型,培养4～5d呈亚融合状态后用于实验.取16孔心肌细胞,按随机数字表法分为空白对照组及1×10-6、1×10-7、1×10-8 mmol/LNE组（NE1、NE2、NE3组）4组,每组4孔；另取12孔心肌细胞,按随机数字表法分为空白对照组、NE3组（1×10-8 mmol/L NE）、T＋NE组（1×10-8 mmol/L NE诱导前30 min加入1×10-5 mmol/L盐酸曲马多）3组,每组4孔.加入NE后24 h,采用免疫组化染色技术测定NF-κB的表达,采用流式细胞仪测定心肌细胞胞质NF-κB的活化程度.结果 免疫组化结果显示,NE3组细胞核周围和核内NF-κB阳性表达较空白对照组增多,NF-κB的平均吸光度（A）值和表达量[阳性单位（PU）]均明显高于空白对照组（A值：0.30±0.03比0.12±0.04,t=7.200,P=0.008；PU：39±14比22±6,t=4.610,P=0.020）.流式细胞仪检测显示,NE1组、NE2组、NE3组细胞质中NF-κB水平均明显低于空白对照组[（45.8±1.9）％、（38.3±1.8）％、（20.9±1.6）％比（54.6±0.6）％,均P＜ 0.05],且NE3组明显低于NE1组、NE2组（均P＜0.05）.T＋NE组细胞质中NF-κB水平明显高于NE3组[（57.8±0.4）％比（20.9±1.6）％,t=57.524,P=0.001].结论 盐酸曲马多预处理可抑制NE诱导大鼠心肌细胞NF-κB的活化,在细胞水平对缺血心肌起保护作用.Objective To investigate the effects of tramadol hydrochloride preconditioning on activation of nuclear factor-κB （NF-κB） induced by norepinephrine （NE） in cultured neonatal rat cardiomyocytes.Methods 1 to 3-day old Sprague-Dawley （SD） rats were raised,and their cardiomyocytes were cultured for 4 to 5 days.Sixteen wells of cultured cardiomyocytes were randomly divided into four groups （n =4）：blank control group,NE 1 group in which the cells were incubated with 1 × 10-6 mmol/L of NE,NE2 group in which cells were incubated with 1 × 10-7 mmol/L of NE,and NE3 group in which cells were incubated with 1 × 10-8 mmol/L of NE.Another 12 wells of cultured cardiomyocytes were randomly divided into three groups （n＝4）：blank control group,NE3 group in which cells were incubated with 1 × 10-8 mmol/L of NE,tramadol plus NE group （T ＋ NE group） in which cells were incubated with 1 × 10-5 mmol/L of tramadol hydrochloride plus 1 × 10-8 mmol/L of NE.Tramadol hydrochloride was added 30 minutes before the use of NE in T ＋ NE group.After exposure to NE for 24 hours,the expression and activation of NF-κB in the cultured cardiomyocytes were determined by immunocytochemistry and flow cytometry （FCM）.Results The results of immunocytochemistry showed that the expression of NF-κB （including average optical density and positive units） in NE3 group was significantly increased as compared with that in the blank control group （A average optical density：0.30 ±0.03 vs.0.12 ±0.04,t=7.200,P=0.008; positive units：39 ± 14 vs.22 ±6,t=4.610,P=0.020）.The results of FCM showed that the levels of cytoplasm NF-κB in cultured cardiomyocytes were decreased significantly in NE1 group,NE2 group and NE3 group compared with blank control group [（45.8 ± 1.9）％,（38.3 ± 1.8）％,（20.9 ± 1.6）％ vs.（54.6 ± 0.6）％,all P＜0.05],and the level in NE3 group was significantly lower than that in NE1 and NE2 groups （both P＜0.05）.The level of the cardiomyocytes cytoplasm NF-κB in T �

关 键 词：盐酸曲马多 心肌细胞 去甲肾上腺素 核转录因子-ΚB 

分 类 号：R614[医药卫生—麻醉学]