法医学杂志

法医学杂志 ›› 2014, Vol. 30 ›› Issue (4): 245-249.DOI: 10.3969/j.issn.1004-5619.2014.04.002

• 论著 • 上一篇    下一篇

不同温度下大鼠皮肤5种RNA指标与PMI的相关性

潘  晖1，张  恒1，吕叶辉1，马剑龙1，马开军2，陈  龙1   

  1. （1. 复旦大学上海医学院法医学系，上海200032； 2. 上海市公安局物证鉴定中心 法医物证学现场应用技术公安部重点实验室 上海市现场物证重点实验室，上海 200083）
  • 发布日期:2014-08-25 出版日期:2014-08-28
  • 通讯作者: 陈龙，男，博士，主任法医师，副教授，主要从事法医病理学研究及法医临床学研究；E-mail：chenlong@shmu.edu.cn
  • 作者简介:潘晖（1988—），女，山东青岛人，硕士研究生，主要从事法医病理学研究；E-mail：12211010056@fudan.edu.cn
  • 基金资助:

    国家自然科学基金资助项目（81172896，81373242）

Correlation between Five RNA Markers of Rat’s Skin and PMI at Different Temperatures

PAN HUI1, ZHANG HENG1, L?譈 YE-HUI1, MA JIAN-LONG1, MA KAI-JUN2, CHEN LONG1   

  1. （1. Department of Forensic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China; 2. Shanghai Key Laboratory of Crime Scene Evidence, Key Laboratory of Forensic Evidence and Science Technology, Ministry of Public Security, Institute of Forensic Science, Shanghai Public Security Bureau, Shanghai 200083, China）
  • Online:2014-08-25 Published:2014-08-28

被引次数

5

摘要： 目的 探讨大鼠皮肤内β-肌动蛋白（β-actin）、甘油醛-3-磷酸脱氢酶（glyceraldehyde-3-phosphate dehydrogenase，GAPDH）、18S核糖体RNA（18S ribosomal RNA，18S rRNA）、5S核糖体RNA（5S ribosomal RNA，5S rRNA）以及微小RNA-203（microRNA-203，miR-203）这5种RNA指标在不同温度下的表达水平与死亡时间（PMI）的相关性。 方法 将18只SD大鼠随机分为3组，处死后分别置于4 ℃、15 ℃和35 ℃的环境中，于死后0~120 h内11个时间点取大鼠腹部皮肤。抽提皮肤总RNA，利用实时荧光定量PCR技术检测5种RNA表达水平。运用geNorm软件选取合适内参后，利用GraphPad软件对内参标准化的RNA指标进行回归分析。 结果 5S rRNA和miR-203作为内参最合适。在4 ℃和15 ℃温度组中，β-actin和GAPDH的表达量变化与PMI线性关系良好。在35 ℃下，β-actin和GAPDH与PMI呈现S形曲线关系。而18S rRNA只在15 ℃和35 ℃温度组呈现一定的线性关系。 结论 皮肤组织比较适合作为提取RNA的检材，其中β-actin和GAPDH表达水平与PMI相关性良好，有望成为推测PMI的辅助指标。

关键词: 法医病理学, RNA, 皮肤, 温度, 死亡时间, 大鼠

Abstract: Objective To explore the correlation between postmortem interval （PMI） and five RNA markers of rat’s skin—β-actin, glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, 18S ribosomal RNA（18S rRNA）, 5S ribosomal RNA （5S rRNA）, and microRNA-203 （miR-203）, at different temperatures. Methods Eighteen SD rats were randomly divided into three environmental temperature groups： 4 ℃, 15 ℃ and 35 ℃, respectively. Skin samples were taken at 11 time points from 0 h to 120 h postmortem. The total RNA was extracted from the skin samples and the five RNA levels were detected by real-time fluorescent quantitative PCR. Proper internal reference was selected by geNorm software. Regression analysis of the RNA markers was conducted by GraphPad software. Results 5S rRNA and miR-203 were most suitable internal references. A good linear relationship between PMI and RNA levels （β-actin and GAPDH） was observed in two groups （4 ℃ and 15 ℃）, whereas the S type curve relationship between the expression levels of the two markers （β-actin and GAPDH） and PMI was observed in the 35 ℃ group. The partial linear relationship between 18S rRNA and PMI was observed in the groups （15 ℃ and 35 ℃）. Conclusion Skin could be a suitable material for extracting RNA. The RNA expression levels of β-actin and GAPDH correlate well with PMI, and these RNA markers of skin tissue could be additional indice for the estimation of PMI.

Key words: forensic pathology, RNA, skin, temperature, postmortem interval, rats

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