摘要
目的 观察吗啡预先给药对去甲肾上腺素(NE)诱导大鼠心肌细胞NF-κB活化的影响,探讨吗啡在细胞水平对缺血心肌的保护作用机制。方法 出生1-3d的SD大鼠,分离、培养心肌细胞,将培养4~5d呈亚融合状态下的细胞随机分为5组(n:4):对照组(C组)、10^-6mol·L^-1NE组(NE1组)、10^-7mol·L^-1 NE组(NE2组)、10^-8mol·L^-1 NE组(NB组)、10^-5mol·L^-1吗啡+10^-8mol·L^-1 NE组(M+NE组)。NE1组、NE2组、NB组细胞培养基内分别加入相应浓度的NE,M+NE组在加入NE前30min加入10^-5mol·L^-1吗啡。加入NE后24h,进行下述指标的测定,采用流式细胞术测定心肌细胞胞浆NF-κB表达水平;采用免疫细胞化学染色技术测定NF-κB、TNF-α表达,采用酶免疫试验测定TNF-α含量,采用反转录PCR技术测定TNF-αmRNA表达。结果 与C组相比,NE1组、NE2组、NE3组细胞浆NF-κB水平降低,其中NB组NF-κB表达水平最低,NR组、NE3组TNF-α含量及TNF-αmRNA表达升高,陋组NF-κB、TNF-α表达升高(P〈0.05);与NE3组比较,M+NE组细胞浆NF-κB水平升高,TNF-α含量及TNF-αmRNA表达降低(P〈0.05)。结论 10^-5mol·L^-1吗啡预先给药可在一定程度上抑制NE诱导大鼠心肌细胞NF-κB的活化,从而减少了TNF-α的释放。
Objective To investigate the effects of morphine pretreatment on norepinephrine (NE)-induced activation of NF-κB in cultured neonatal rat cardiomyocytes. Methods The cultured rat cardiomyocytes were obtained from 1-3 day old SD rats and cultured for 4-5 days. The cells were randomly divided into 5 groups : group A control; group B NE 10^-6mol·L^-1(NE1); group C NE 10^-7 mol·L^-1(NF2); group D NE 10^-8 mol·L^-1 (NE3 ) and group E morphine 10-5 mol·L^-1 + NE 10^-8mol·L^-1(M + NE). In group E morphine was added to the culture media 30min before NE. The following measurements were made 24 h after NE had been added: ( 1 ) percentage of NF-κB expression in the cytoplasm of cardiomyocytes indicating the degree of activation of NF-κB (flow cytometry ) ; ( 2 ) expression of NF-κB and TNF-α ( immuno-histochemistry ) ; ( 3 ) TNF-α concentration (enzyme immunometric assay EIA) and TNF-α mRNA expression (RT-PCR).Results The expression and the degree of activation of NF-κB and the level of TNF-α were significantly increased in the 3 NE groups ( NE1. 2.3 ) dose-dependently as compared with control group and group E (M + NE). Conclusion Morphine pretreatment can inhibit NE-induced activation of NF-κB in cardiomyocytes and in turn decrease TNF-α release.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2007年第1期66-69,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金资助项目(30471656)