摘要
目的 探讨烟碱对心肌缺血/再灌注（I/R）损伤大鼠炎症细胞因子的影响.方法 50只健康雄性SD大鼠按随机数字表法分为假手术组、I/R组、烟碱高剂量（400μg/kg）组、烟碱低剂量（40μg/kg）组及α-银环蛇毒素（α-BGT,1μg/kg）组5组,每组10只.采用结扎心脏左冠状动脉前降支30 min、再灌注90 min制作大鼠心肌I/R损伤模型;假手术组仅穿线不结扎.制模前30 min各药物组颈静脉注射相应剂量药物干预,假手术组和I/R组给予等量生理盐水.于再灌注末取右颈动脉血,测定肿瘤坏死因子-α（TNF-α）、白细胞介素-8（IL-8）、IL-10浓度和肌酸激酶同工酶（CK-MB）、心肌肌钙蛋白I（cTnI）活性;然后处死动物,取缺血区心肌组织测定髓过氧化物酶（MPO）活性;采用免疫组化和逆转录-聚合酶链反应检测心肌组织细胞间黏附分子-1（ICAM-1）蛋白及mRNA表达,并观察心肌超微结构.结果 与假手术组比较,I/R组血浆TNF-α、IL-8、IL-10、CK-MB、cTnI、心肌MPO活性及ICAM-1蛋白和mRNA表达均显著升高[TNF-α（ng/L）：158.7±32.7比31.5±5.8,IL-8（ng/L）：0.71±0.06比0.30±0.04,IL-10（ng/L）：69.0±7.8比41.4±4.3,CK-MB（U/L）：2 540±169比1 120±102,cTnI（μg/L）：26.2±4.6比0.9±0.2,MPO（U/g）：4.2±0.6比1.6±0.4,ICAM-1蛋白：0.210±0.025比0.100±0.018,ICAM-1 mRNA：1.82±0.23比1.18±0.20,P＜0.05或P＜0.01],病理学显示心肌组织损伤较重.与I/R组比较,烟碱高剂量组血浆TNF-α、IL-8降低[TNF-α（67.3±9.8）ng/L,IL-8（0.47±0.04）ng/L],IL-10升高[（147.5±12.5）ng/L],CK-MB、cTnI及心肌MPO活性、ICAM-1蛋白和mRNA均降低[CK-MB（1 282±145）U/L,cTnI（4.7±1.4）μg/L,MPO（2.5±0.4）U/g,ICAM-1蛋白0.140±0.026,ICAM-1 mRNA 1.31±0.25,P＜0.05或P＜0.01],心肌组织损伤减轻;而烟碱低剂量组和α-BGT组上述指标与I/R组比较差异无统计学意义.结论 烟碱可阻断内皮细胞表达黏附分子,阻断中性粒细胞黏附、游出,改善抗炎/促炎反应平衡,从而拮抗大鼠心肌I/R损伤时的过度炎症反应.
Objective To investigate the effect of nicotine on inflammatory cytokines in myocardial ischemia/reperfusion （I/R） injury in rat. Methods Fifty male Sprague-Dawley （SD） rats were divided into five groups by random numbers table （each n= 10）： sham operation group （S group）, I/R group, nicotine 400 μg/kg group （H group）, nicotine 40 μg/kg group （L group） and α-bungarotoxin （α-BGT, 1 μg/kg）group. The anterior descending branch of left coronary artery was occluded for 30 minutes followed by 90 minutes reperfusion to reproduce myocardial I/R injury rat model, while in S group the anterior descending branch of left coronary artery was only exposed without occlusion procedure. Thirty minutes before myocardial ischemia, drugs in corresponding doses were given intravenously via jugular vein. At the end of 90 minutes of reperfusion, blood samples were collected from carotid artery to determine the levels of tumor necrosis factor-α （TNF-α）, interleukin-8 （IL-8）, IL-10, MB isoenzyme of creatine kinase （CK-MB）,and cardiac troponin I （cTnI）, then the animals were sacrificed and the hearts were harvested for pathological study and determination of myeloperoxidase （MPO） activity. Immunohistochemistry and reverse transcription-polymerase chain reaction （RT-PCR） were used to assess intercellular adhesion molecule-1 （ICAM-1） protein and mRNA expression in heart tissue. Results Compared with the S group, the concentrations of TNF-α, IL-8, IL-10, CK-MB, cTnI, MPO activity, ICAM-1 protein and mRNA expression were significantly increased in I/R group [TNF-α （ng/L）： 158. 7± 32. 7 vs. 31.5±5.8, IL-8（ng/L）： 0.71±0.06 vs. 0. 30±0. 04, IL-10 （ng/L）： 69.0±7.8 vs. 41.4±4.3, CK-MB （U/L）： 2540±169 vs. 1 120±120, cTnI （μg/L）： 26.2±4.6 vs. 0.9±0.2, MPO （U/g）： 4.2±0.6 vs. 1.6±0.4,ICAM-1 protein： 0. 210±0. 025 vs. 0. 100±0. 018, ICAM-1 mRNA： 1.82±0.23 vs. 1.18±0. 20, P〈0. 05or P〈0. 01]. Injury to myocardial ultrastructure was worse in I/R group. Compared with the I/R group,the plasma levels of TNF-α and IL-8 were lower [TNF-α （67.3±9.8） ng/L, IL-8 （0. 47±0. 04） ng/L],IL-10 was higher [（147.5± 12.5） ng/L], CK-MB, cTnI, MPO, ICAM-1 protein and mRNA were lower obviously in H group [CK-MB （1 282±145） U/L, cTnI （4. 7±1.4） μg/L, MPO （2. 5±0. 4） U/g, ICAM-1 protein 0. 140 ± 0. 026, ICAM-1 mRNA 1.31 ± 0.25, P 〈 0. 05 or P 〈 0. 01]. Injury to the myocardial ultrastructure was less marked in H group. The indexes of those in L group and α-BGT group compared with I/R group were not statistically significantly different. Conclusion Nicotine can block endothelial expression of adhesion molecules and neutrophil adhesion and infiltration to promote a balance of anti-inflammatory and pro-inflammatory response, thus prevents excessive inflammatory response to myocardial I/R injury in rat.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2010年第10期624-627,I0002,共5页
Chinese Critical Care Medicine
基金
基金项目：湖北省武汉市医药卫生资助项目（WX09A04）
关键词
烟碱
缺血/再灌注损伤
心肌
胆碱能抗炎通路
炎症细胞因子
大鼠
Nicotine
Myocardial ischemia/reperfusion injury
Cholinergic anti-inflammatory pathway
Inflammatory cytokine
Rat
