【摘要】 目的评价miR-499诊断急性心肌梗死的法医学应用价值。方法利用实时定量PCR技术,研究miR-499诊断急性心肌梗死的敏感性、死后稳定性及鉴别不同死因的能力。首先,在急性心肌梗死发生后不同时间,包括心梗即刻、心梗0.25、0.5、1、2、3、6、12h以及心梗7d,采用real-time PCR技术检测心血血浆中miR-499表达变化;其次,心梗1h死亡和断颈死亡后,分别在死亡即刻及3、6、12、24h采集心血检测miR-499的相对表达量;最后,将大鼠分为断颈死亡、窒息死亡、麻醉过量死亡以及心梗1h死亡组,采用实时荧光定量PCR方法检测miR-499的表达量。结果心梗1h,mi R-499的相对表达倍数达到峰值,是心梗即刻的(64.90±9.26)倍,并于心梗7d逐渐回落至与心梗即刻持平;不同死亡时间中,心梗1h死后miR-499的相对表达倍数及变化幅度均大于断颈死亡组,且在死后3h心梗组miR-499的相对表达倍数是断颈死亡组的3.01倍;在不同死亡原因中,心梗1h的△CT值高于其他死因,且心梗1h与麻醉死亡及窒息死亡的△CT值存在明显差异(P <0.01)。结论 miR-499用于急性心肌梗死的法医学诊断存在巨大潜力。更多还原

【Abstract】 Objective To evaluate the forensic value of miR(microRNA)-499 to diagnose acute myocardial infarction. Methods Real-time quantitative PCR(RT-qPCR) was conducted to explore the diagnostic sensitivity and postmortem stability of miR-499 in acute myocardial infarction and the discriminability of different causes of death. With the RT-qPCR to detect the miR-499 at different infarction interval, i.e., the moments of both the instantaneous and post-myocardial infarction at, 15 min, 30 min, 60 min and 2, 3, 6, 12 hours and 7 days, the experimental rats were taken of their heart blood to get known of the miR-499 expressions. Similarly, the rats, died from either the acute myocardial infarction lasting for 1 hour or broken neck, had their heart blood also taken at different postmortem interval(the moment of immediate death and after hours of 3, 6, 12, 24) to detect the miR-499 expressions. Finally, the experimental rats were divided into 4 groups, respectively killed with suffering for 1 hour myocardial infarction, neck breaking, suffocating and anesthetizing, having their miR-499 expressions detected through RT-qPCR. Results The miR-499 got peaked of relative expression multiplication at 1 hour myocardial infarction, showing 64.90±9.29 times of that at the immediate myocardial infarction which to be gradually fallen back from the peak until after 7 days. In different postmortem intervals, miR-499 was obviously greater at the death from 1 hour myocardial infarction than that from the neck breaking group in relation to its variation range and the relative expression multiplication. For 3 hours after death, miR-499 was 3.01 times higher of the myocardial infarction group than that of the broken neck group. In different causes of death, △ CT was higher of 1 hour myocardial infarction than that of the other death cause, demonstrating signi? cant difference among the 1 hour myocardial infarction, suffocation and anesthesia(P<0.01). Conclusion The miR-499 expression is of great potential in forensic diagnosis for acute myocardial infarction.更多还原