摘要
目的研究离体肝脏缺血再灌注早期丝裂原活化蛋白激酶(mitogen activated protein kinase,p38MAPK)信号转导途径对肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)mRNA和细胞间黏附分子-1(inter-cellular adhesion molecule1,ICAM1)mRNA表达的影响。方法建立兔离体肝脏缺血再灌注模型,对照组(n=12):灌注液中不加特异性p38MAPK抑制剂SB202190;抑制组(n=12):灌注液中加入SB202190(浓度3μmol/L)。分别于肝脏离体前,冷保存末,再灌注10、30、60及120min时获取肝组织标本。分别应用Western blot法及免疫沉淀法检测肝组织中p38MAPK蛋白的表达及活性;RT-PCR法检测肝组织中TNF-α mRNA的表达水平,原位杂交法检测肝组织中ICAM1 mRNA的表达水平。结果2组动物肝组织中p38MAPK蛋白的表达水平各时相均无明显改变(P>0.05),且2组间其表达水平的差异也无统计学意义(P>0.05)。对照组肝组织中p38MAPK的活性在冷保存末及再灌注10、30及60min时均较离体前和再灌注120min时明显升高(P<0.01),也明显高于同时相的抑制组(P<0.01);抑制组p38MAPK活性各时相的变化差异无统计学意义(P>0.05)。离体前、冷保存末及再灌注10及30min,2组的肝组织中均仅有少量TNF-α mRNA和ICAM1 mRNA表达,组间及组内比较差异均无统计学意义(P>0.05);至再灌注60及120min,2组TNF-α mRNA和ICAM1 mRNA的表达水平均明显升高(P<0.05,P<0.01),但抑制组相应时相的表达水平明显低于对照组(P<0.01)。离体再灌注期间肝组织中p38MAPK的活性与TNF-α mRNA及ICAM1 mRNA的表达水平呈正相关(r=0.996,P<0.01;r=0.985,P<0.01)。结论p38MAPK可能是在转录水平对TNF-α和ICAM1的生成发挥调节作用,p38MAPK信号转导途径对TNF-α mRNA和ICAM1 mRNA的调节可能是导致离体肝脏缺血再灌注损伤的重要机理之一。
Objective To study the effect of p38MAPK activity on tumor necrosis factor-α （TNF-α） mRNA and intercellular adhesion molecule 1 （ICAM1） mRNA expressions of isolated rabbit liver during early stage of cold preservation and reperfusion period. Methods Based on the cold preservation and reperfusion model of isolated rabbit liver, the animals were divided into inhibition group （n= 12） with 3 μmol/L SB202190 （p38MAPK specificity inhibitor） in perfusate and control group （n= 12） with no SB202190 in perfusate. Liver tissue samples were harvested at the time points of before resection, end of cold preservation, and different reperfusion period （10, 30, 60 and 120 vain）. Protein expression and activity of p38MAPK were detected by Western blot and immunoprecipitation respectively, expression of TNF- α mRNA was detected by RT-PCR, and expression of ICAM1 mRNA was detected by in situ hybridization. Results There was no obvious change of expression of p38MAPK protein in liver tissue both in two groups during the total period （P〉0.05）, and there was no statistically significant difference between two groups （P〉0.05）. At time points of end of cold preservation, 10, 30 and 60 rain of reperfusion, the activity of p38MAPK in control group was significantly higher than that at the time points of before resection and 120 min of reperfusion （P〈0. 01）, and was also significantly higher than that in inhibition group at the same time points （P〈0.01）. There was no significant difference in activity of p38MAPK among all time points in inhibition group （P〉0.05）. The expressions of TNF-α mRNA and ICAM1 mRNA at the time points of before resection, end of cold preservation, and 10 and 30 min of reperfusion were significantly lower than those in 60 and 120 min of reperfusion in both two groups （P〈0.05, P〈0.01） ; The expressions of TNF-α mRNA and ICAM1 mRNA in inhibition group were significantly lower than those in control group at the time points of 60 and 120 min of reperfusi
出处
《中国普外基础与临床杂志》
CAS
2009年第6期443-448,共6页
Chinese Journal of Bases and Clinics In General Surgery
关键词
肝
缺血再灌注
损伤
丝裂原活化蛋白激酶
肿瘤坏死因子
细胞间黏附分子
抑制剂
Liver
Ischemia-reperfusiom Injury
Mitogen activated protein kinase
Tumor necrosis factor
Intercellular adhesion molecule
Inhibitor
