骨髓相关蛋白在异丙肾上腺素制作的大鼠心肌损伤坏死模型中的诊断价值
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摘要
目的:在探索大鼠心肌损伤坏死模型中骨髓相关蛋白(MRP8/14)的变化的基础上,进一步了解MRP8/14在心肌坏死事件中的诊断价值。方法:首先检测健康成年Sprauge Dawley大鼠Ⅱ导联心电图,剔除异常心电图者,适应性饲养1周。把大鼠分成两组:模型组,16只;对照组,10只。模型组大鼠给予一次性多点皮下注射异丙肾上腺素10mg/kg,每隔24h注射1次,连续3次。给予对照组大鼠等量0.9%氯化钠。于第3次给药后1、3、6、8h经左侧颈总动脉采动脉血,用分光光度计测定血清中肌钙蛋白(cTnI)、MRP8/14含量,末次取血后立即打开大鼠胸腔取出心脏,并制作病理切片,在光镜下观察心肌形态学变化。结果:①光镜下发现模型组心肌组织坏死、肌纤维断裂、肌细胞水肿、大量炎性细胞浸润。②MRP8/14:模型组与对照组不满足球形对称,校正后两组差别具有统计学意义(P<0.001),不同时间点总体间差异具有统计学意义(P<0.001),且时间与分组存在交互效应(P<0.001)。对照组与模型组比较:在1h时间点两组比较差异无统计学意义,在3h、6h、8h时间点两组比较差异有统计学意义。结论:在异丙肾上腺素制作大鼠心肌损伤坏死模型中,与cTnI类似,MRP8/14可作为该模型的诊断指标,较cTnI更早期反映心肌坏死的发生。
Objective:To study the value of diagnosis of myeloid-related protein in the event of myocardial necrosis,through exploring the changes of myeloid-related protein in rat myocardial ischemia and necrosis model. Method:The rats whose ECG were normal were divided into two groups:control group(n=10)and model group(n=16).The rats in model group were given the disposable multi subcutaneous injection of isoproterenol 10mg/ kg,inject once every 24hours,three times in succession.The rats in control group were given the same dose of saline.We took blood 0.5ml from left carotid artery at four different time points:1hour,3hours,6hours,8 hours,after the last injection.The change of cTnI,MRP8/14in blood serum were examined by model visible-infrared spectromer after the last blood obtaining,then opened the rats' chest,got the heart at once,washed it with 0.9% NaCl and removed the big vessel and kept it in formalin solution for making HE histopathology slices. Result:①Under light microscopic,we observed that there were myocardial tissue patchy necrosis,myocardial fiber breakage,myocardial cell edema and inflammatory cell filtration in control group.②MRP8/14:The expressions of MRP8/14in two groups did not satisfied sphericity test.After correction,the difference between the two groups was statistically significant(P<0.001).There were significantly differences among different time points overall between two groups(P<0.001).There was interaction between time and grouping.Compared with control group,there were significant differences at 3hours,6hours and 8hours point in model group.Conclusion: Myeloid-related protein 8/14could be used as a diagnostic indicator of rats myocardial necrosis produced by isoproterenol,as well as troponin,even earlier than troponin rised when myocardial necrosised.
Objective:To study the value of diagnosis of myeloid-related protein in the event of myocardial necrosis,through exploring the changes of myeloid-related protein in rat myocardial ischemia and necrosis model. Method:The rats whose ECG were normal were divided into two groups:control group(n=10)and model group(n=16).The rats in model group were given the disposable multi subcutaneous injection of isoproterenol 10mg/ kg,inject once every 24hours,three times in succession.The rats in control group were given the same dose of saline.We took blood 0.5ml from left carotid artery at four different time points:1hour,3hours,6hours,8 hours,after the last injection.The change of cTnI,MRP8/14in blood serum were examined by model visible-infrared spectromer after the last blood obtaining,then opened the rats' chest,got the heart at once,washed it with 0.9% NaCl and removed the big vessel and kept it in formalin solution for making HE histopathology slices. Result:①Under light microscopic,we observed that there were myocardial tissue patchy necrosis,myocardial fiber breakage,myocardial cell edema and inflammatory cell filtration in control group.②MRP8/14:The expressions of MRP8/14in two groups did not satisfied sphericity test.After correction,the difference between the two groups was statistically significant(P<0.001).There were significantly differences among different time points overall between two groups(P<0.001).There was interaction between time and grouping.Compared with control group,there were significant differences at 3hours,6hours and 8hours point in model group.Conclusion: Myeloid-related protein 8/14could be used as a diagnostic indicator of rats myocardial necrosis produced by isoproterenol,as well as troponin,even earlier than troponin rised when myocardial necrosised.
引文
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[9]VORA A N,BONACA M P,RUFF C T,et al.Diagnostic evaluation of the MRP-8/14for the emergency assessment of chest pain[J].J Thromb Thrombolysis,2012,34:229-234.
[10]DEKKER M S,MOSTERD A,VAN'T HOF A W,et al.Novel biochemical markers in suspect-acute cor-onary syndrome:systematic review and critical appraisal[J].Heart,2010,96:1001-1010.
[11]SZEKANECZ Z,KOCH A E.Analysis of inflammatory leukocyte and endothelial chemotactic activity[J].Methods Mol Med,2007,135:359-364.
[12]SEELIGER S,VOGL T,ENGELS I H,et al.Expression of calcium-binding proteins MRP8 and MRP14in inflammatory muscle diseases[J].Am J Pathol,2003,163:947-956.
[13]HAN H G,WANG Z W,ZHANG N B,et al.Role of nitric oxide during early phase myocardial ischemic preconditioning in rats[J].Chin Med J,2008,121:1210-1214.
[14]WALLACE K B,HAUSNER E,HERMAN E,et al.Serum troponins as biomarkers of drug-induced cardiac toxicity[J].Toxicol Pathol,2004,32:106-121.
[2]ALTWEGG L A,NEIDHART M.Myeloid-related protein8/14complex is released by monocytes and granulocytes at the site of coronary occlusion:a novel,early,and sensitive marker of acute coronary syndromes[J].Eur Heart J,2007,28:941-948.
[3]STRONCEK D F,SHANKAR R A,SKUBITZ K M.The subcellular distribution of myeloid-related protein 8(MRP8)and MRP14in human neutrophils[J].J Transl Med,2005,3:36-40.
[4]CROCE K,GAO H,WANG Y,et al.Myeloid-related protein-14is cirtical for the biological response to vascular injury[J].Circulation,2009,120:427-436.
[5]IONITA M G,VINK A,DIJKE I E,et al.High levels of myeloid-related protein 14in human atherosclerotic plaques correlate with the characteristics of rup ture-prone lesions[J].Arterioscler Thromb VascBiol,2009,29:1220-1227.
[6]MAISEYEU A,BADGELEY M A,KAMPFRATH T,et al.In vivo targeting of inflammation associated myeloid-related protein 8/14via gadolinium immunonanoparticles[J].Arterioscler Thromb Vasc Biol,2012,32:962-970.
[7]YONEKAWA K,NEIDHART M,ALTWEQQ M,et al.Myeloid related proteins activate Toll-like receptor 4in human acute coronary syndromes[J].Atherosclerosis,2011,218:486-492.
[8]张永军,汤圣兴,等.生化标志物联合检测对急性冠脉综合征早期诊断的意义[J].皖南医学院学报,2011,30(5):365-367.
[9]VORA A N,BONACA M P,RUFF C T,et al.Diagnostic evaluation of the MRP-8/14for the emergency assessment of chest pain[J].J Thromb Thrombolysis,2012,34:229-234.
[10]DEKKER M S,MOSTERD A,VAN'T HOF A W,et al.Novel biochemical markers in suspect-acute cor-onary syndrome:systematic review and critical appraisal[J].Heart,2010,96:1001-1010.
[11]SZEKANECZ Z,KOCH A E.Analysis of inflammatory leukocyte and endothelial chemotactic activity[J].Methods Mol Med,2007,135:359-364.
[12]SEELIGER S,VOGL T,ENGELS I H,et al.Expression of calcium-binding proteins MRP8 and MRP14in inflammatory muscle diseases[J].Am J Pathol,2003,163:947-956.
[13]HAN H G,WANG Z W,ZHANG N B,et al.Role of nitric oxide during early phase myocardial ischemic preconditioning in rats[J].Chin Med J,2008,121:1210-1214.
[14]WALLACE K B,HAUSNER E,HERMAN E,et al.Serum troponins as biomarkers of drug-induced cardiac toxicity[J].Toxicol Pathol,2004,32:106-121.