摘要
目的:建立近似人类新生儿窒息的病理过程的新生鼠窒息心肌损伤模型,观察心肌HO-1 mRNA、NF-кB表达关系及血中CK-MB、cTnI的变化。方法:正常对照组8只,窒息组80只(分为窒息组40只,窒息+ZnPP组40只)。又分为窒息后6 h,24 h,48 h,72 h,7天,5个不同的时间段采血和心肌样本。①电镜下观察心肌组织细微病变;②原位杂交技术检测HO-1 mRNA在心肌组织的表达;③免疫组化技术检测NF-кB在心肌组织的表达;④ELISA技术检测血中CK-MB、cTnI水平。结果:窒息组:电镜下各时段心肌组织均有线粒体肿胀;核固缩,肌浆网肿胀;窒息+ZnPP组:电镜下各时段均有心肌细胞融合,肌丝溶解,线粒体肿胀,线粒体脊溶解。窒息+ZnPP组的病理损伤比窒息组严重。窒息组血中CK-MB,cTnI水平明显高于对照组,有统计学差异(P<0.01);窒息+ZnPP组血中CK-MB,cTnI水平也明显高于对照组,差异有统计学意义(P<0.01);窒息组血中CK-MB,cTnI水平明显低于窒息+ZnPP组,差异有统计学意义(P<0.05)。窒息组心肌组织细胞HO-1 mRNA表达高于对照组,差异有统计学意义(P<0.01);窒息+ZnPP组心肌细胞HO-1 mRNA表达高于对照组,差异有统计学意义(P<0.05);窒息+ZnPP组与窒息组比较,HO-1 mRNA表达明显降低,差异有统计学意义(P<0.01)。窒息组心肌组织细胞NF-кB表达高于对照组,差异有统计学意义(P<0.01);窒息+ZnPP组心肌细胞NF-кB表达高于对照组,差异有统计学意义(P<0.01);窒息+ZnPP组与窒息组比较,NF-кB表达明显增高,差异有统计学意义(P<0.05)。结论:①新生鼠窒息后心肌组织和细胞受到明显的损伤,CK-MB,cTnI在外周血中明显升高,心肌细胞融合,肌丝溶解,线粒体肿胀;②外周血中CK-MB持续时间较短,cTnI在血中持续时间较长,约7天,提示对心肌损伤的防治应持续至窒息后7天,以免加重心肌损伤和后遗症发生;③HO-1 mRNA表达与NF-кB表达呈负相关;④提示在窒息致心肌损伤的防治中,可适时使用HO-1 mRNA诱导剂,减少炎性细胞因子对心肌的损伤。
Objective：To establish an animal model of myocardial injury induced by asphyxia similar to the pathological course of myocardial injury induced by human neonatal asphyxia,observe the relationship between heme oxygenase-l（HO-1） mRNA expression and NF-кB expression and the changes of CK-MB,cTnI in blood.Methods：88 SD rats were selected and divided into control group（n=8）,asphyxia group（n=40） and asphyxia＋ZnPP group（n=40）.The blood samples and myocardial samples were obtained at 6 hours,24 hours,48 hours,72 hours and 7 days after asphyxia.The minimal lesions of myocardial tissues were observed under electron microscope;in situ hybridization technique was used to detect the expression level of HO-1 mRNA in myocardial tissues;the expression level of NF-кB in myocardial tissues were detected by Immunohistochemistry;ELISA technique was used to detect the expression levels of CK-MB,cTnI in blood.Results：Asphyxia group： at all time periods,swelling of mitochondria,karyopyknosis and swelling of sarcoplasmic reticulum in myocardial tissues were observed under electron microscope.Asphyxia＋ZnPP group： at all time periods,fusion of myocardial cells,solution of myofilaments,swelling of mitochondria and solution of mitochondria cristae in myocardial tissues were observed under electron microscope.The pathological injury in asphyxia＋ZnPP group was more severer than that in asphyxia group.The expression levels of CK-MB,cTnI in blood in asphyxia group were significantly higher than those in control group（P0.01）;the expression levels of CK-MB,cTnI in blood in asphyxia＋ZnPP group were significantly higher than those in control group（P0.01）;the expression levels of CK-MB,cTnI in blood in asphyxia group were significantly lower than those in asphyxia＋ZnPP group（P0.05）.The expression level of HO-1 mRNA in myocardial cells in asphyxia group was significantly higher than that in control group（P0.01）;the expression level of HO-1 mRNA in myocardial cells in asphyxia＋ZnPP group was significantly higher than that in control group（P0.05）;the expression level of HO-1 mRNA in myocardial cells in asphyxia＋ZnPP group was significantly lower than that in asphyxia group（P0.01）.The expression level of NF-кB in myocardial cells in asphyxia group was significantly higher than that in control group（P0.01）;the expression level of NF-кB in myocardial cells in asphyxia＋ZnPP group was significantly higher than that in control group（P0.01）;the expression level of NF-кB in myocardial cells in asphyxia＋ZnPP group was significantly higher than that in asphyxia group（P0.05）.Conclusion：After asphyxia,the myocardial tissues and cells of neonatal rats are damaged obviously,the expression levels of CK-MB and cTnI in peripheral blood increase significantly,fusion of myocardial cells,solution of myofilaments and swelling of mitochondria can be observed;the duration time of CK-MB in peripheral blood is short,the duration time of cTnI in peripheral blood is long,about 7 days,the phenomena indicate that the prevention and treatment of myocardial injury should continue till 7 days after asphyxia,in order to avoid the deterioration of myocardial injury and the occurrence of sequelae;there is a negative correlation between HO-1 mRNA expression and NF-кB expression;during the prevention and treatment of myocardial injury,appropriate application of HO-1 mRNA inducer can reduce myocardial injury induced by inflammatory factors.
出处
《中国妇幼保健》
CAS
北大核心
2011年第14期2180-2184,共5页
Maternal and Child Health Care of China
基金
云南省面上基金资助项目〔20107M203C〕
