乳化异氟烷预处理对大鼠心肌缺血再灌注时TLR4-α、NF-κB及血清中炎症细胞因子的影响

发布时间：2018-09-09 16:59

【摘要】：目的探讨乳化异氟烷预处理对心肌缺血再灌注损伤可能的保护作用。方法健康雄性SD大鼠48只,体重230~280 g,随机分成4组(n=12):S组(假手术)、IR组(缺血再灌注)、L组(脂肪乳剂预处理)、EI组(乳化异氟烷预处理)。取大鼠(每组6只),于再灌注120 min后采集股动脉血2 m L用于测定血清中心肌钙蛋白I(c Tn I)浓度及肌酸激酶同工酶(CK-MB)活性,实验结束后红四氮唑染色(TTC)法测定心肌梗死范围。另取大鼠(每组6只)于再灌注30 min(T1)、再灌注120 min(T2)时采集股动脉2 m L,采用酶联免疫吸附法(ELISA)测定血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-10(IL-10)的浓度,随后处死大鼠,取心脏,采用荧光定量逆转录聚合酶链反应(Real-time PCR)法测定心肌组织中TLR4 mRNA表达、Western blot法测定心肌细胞核因子-κB(NF-κB)活性,电镜下观察心肌细胞超微结构的变化。结果与S组比较,IR组、L组和EI组心肌细胞TLR4mRNA表达增强,NF-κB活性升高,血清TNF-α、IL-10、c Tn I浓度及CK-MB活性升高,心肌梗死面积增加(P0.05);与IR组比较,EI组心肌细胞TLR4 mRNA表达明显减弱,NF-κB活性降低,血清TNF-α、IL-10、c Tn I浓度及CK-MB活性降低(P0.05),IL-10浓度升高(P0.05),心肌梗死面积减小(P0.05),而L组上述指标差异无统计学意义。电镜结果显示:EI组心肌细胞损伤程度较IR组和L组减轻。结论乳化异氟烷预处理对缺血再灌注心肌具有一定的保护作用,这种保护作用可能与乳化异氟烷抑制TLR4 mRNA表达,降低NF-κB活性,抑制炎性反应有关。
[Abstract]:Objective to investigate the protective effect of emulsified isoflurane preconditioning on myocardial ischemia reperfusion injury. Methods 48 healthy male SD rats, weighing 230 ~ 280g, were randomly divided into 4 groups (n = 12): s group (sham operation) or IR group (ischemia / reperfusion) group (fat emulsion preconditioning) or Ei group (emulsified isoflurane preconditioning). The femoral arterial blood (2 mL) was collected from 6 rats in each group after 120 min reperfusion to determine the concentration of serum troponin I (c Tn I) and the activity of creatine kinase isoenzyme (CK-MB). The myocardial infarction size was measured by (TTC) staining with red tetrazole after the end of the experiment. Six rats in each group were collected 2 mL of femoral artery after reperfusion for 30 min (T1) and 120 min (T2). Serum levels of tumor necrosis factor- 伪 (TNF- 伪) and interleukin-10 (IL-10) were measured by enzyme-linked immunosorbent assay (ELISA). The activity of nuclear factor- 魏 B (NF- 魏 B) in myocardium was determined by fluorescence quantitative reverse transcriptase polymerase chain reaction (Real-time PCR). The ultrastructure of myocardial cells was observed by electron microscope. Results compared with S group, the expression of TLR4mRNA in cardiomyocytes of IR group and EI group increased, the concentration of serum TNF- 伪 -IL-10c Tn I and CK-MB activity increased, and the myocardial infarction area increased (P0.05), and the TLR4 mRNA expression of cardiac myocytes in EI group decreased significantly compared with IR group. Serum TNF- 伪 IL-10c Tn I concentration and CK-MB activity decreased (P0.05), IL-10 concentration increased (P0.05), myocardial infarction area decreased (P0.05), but there was no significant difference in the above indexes in L group. The results of electron microscope showed that the degree of myocardial cell injury in group E was less than that in group IR and group L. Conclusion Emulsion isoflurane preconditioning has a protective effect on ischemic reperfusion myocardium, which may be related to the inhibition of TLR4 mRNA expression, the decrease of NF- 魏 B activity and the inhibition of inflammatory response by emulsified isoflurane.
【作者单位】：西南医科大学附属中医医院麻醉科;
【分类号】：R614

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