人外周血心肌肌钙蛋白Ⅰ基因表达水平与心肌损伤程度的关系  

Relationship between the expression of human peripheral blood myocardial troponin Ⅰgene and myocardial damage

作　　者：姚登福[1] 朱健华[2] 吴玮[1] 高增栋[3] 吴信华[1] 邱历伟[1] 

机构地区：[1]南通医学院附属医院临床分子生物学中心,江苏省南通市226001 [2]南通医学院附属医院心内科,江苏省南通市226001 [3]南通医学院附属医院内科,江苏省南通市226001

出　　处：《中国临床康复》2004年第30期6625-6627,i001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：江苏省自然科学基金项目资助(BK99160)~~

摘　　要：目的:分子生物技术的发展,使监测心肌微小损伤成为可能。探讨外周血心肌肌钙蛋白Ⅰ(cardiactroponinⅠ,cTnⅠ)-mRNA对心肌损伤诊断的临床价值。方法:以电镜观察心肌损伤时的超微结构改变;从心肌组织和外周血中制备总RNA,以随机引物和反转录酶合成cTnⅠ-cDNA,再以巢式PCR扩增和测序分析;将cTnⅠ-mRNA扩增法用于临床诊断,与心肌酶谱(CK,CK-MB,AST,LDH)及cTnⅠ定性比较,以探讨其临床价值。结果:心肌损伤组织超微结构表现为心肌线粒体肿胀、嵴断裂、严重空泡样变及细胞核异形变、染色质浓缩边聚。巢式PCR法扩增cTnⅠ-mRNA片段的灵敏度为2μg/L,测序分析证实从心肌组织及外周血中扩增基因片段与原序列完全一致。对62例慢性心脏病患者外周血cTnⅠ-mRNA的扩增分析,其阳性率明显高于酶谱或cTnⅠ定性分析(P<0.05)。结论:心肌损伤时cTnⅠ-mRNA释放进入外周血,为心肌损伤诊断的敏感基因标志物。AIM:With the development of clinical molecular biology,the measurement of tiny myocardial injury is possible.This article is to investigate the clinical values of peripheral blood cardiac troponin Ⅰ(cTnⅠ) messenger RNA in the diagnosis of myocardial injury.METHODS:Microstructure of myocardial damages was observed under electron microscopy.The cTnⅠ mRNAs were extracted from human heart tissue and peripheral blood and synthesized to cTnⅠ cDNA through ramdom promers and reverse transcriptase,and then cTnⅠ cDNA was amplified by a nested polymerase chain reaction assay.Sequencing analysis of the PCR products was performed,and its clinical value was investigated and compared with heart zymogram(CK,CK MB,AST,LDH) and cTnⅠqualitatively.RESULTS:Microstructure of myocardial injury presented as myocardial mitochondrial swell,crista rupture,severe vacancy like denaturalization and nucleus abnormality and chromatin concentration.The sensitivity of cTnⅠ mRNA segment with nest PCR proliferation method was 2 μg/L,and the amplified gene segment from human myocardial tissues and peripheral blood was in accordance with original sequence by sequencing analysis.The cTnⅠ mRNA amplification analysis used in the peripheral blood of 62 patients with chronic heart disease showed that the positive rate was significantly higher than that of zymogram or cTnl analysis(P< 0.05).CONCLUSION:cTnⅠ mRNA in myocardial cell can release into peripheral blood,which is a sensitive gene marker for the diagnosis of myocardial injury.

关 键 词：心肌损伤 外周血 扩增 诊断 cTnⅠ 心肌肌钙蛋白Ⅰ RNA 基因表达水平 酶合成 巢式PCR法 

分 类 号：R542[医药卫生—心血管疾病] R541[医药卫生—内科学]