法医学杂志

法医学杂志 ›› 2019, Vol. 35 ›› Issue (2): 160-165.DOI: 10.12116/j.issn.1004-5619.2019.02.006

所属专题： 损伤时间推断

• 专题 • 上一篇    下一篇

联合多个修复相关基因的mRNA用于大鼠骨骼肌损伤时间推断

鲁翰霖， 党丽虹， 李娜， 董塔娜， 杜秋香， 王英元， 孙俊红   

  1. 山西医科大学法医学院，山西 太原 030001
  • 发布日期:2019-04-25 出版日期:2019-04-28
  • 通讯作者: 通信作者：孙俊红，男，博士，教授，主要从事损伤病理学和猝死病理学研究；E-mail：junhong.sun@sxmu.edu.cn 通信作者：王英元，男，教授，主要从事损伤病理学和猝死病理学研究；E-mail：wyy580218@163.com
  • 作者简介:鲁翰霖（1991—），男，硕士研究生，主要从事法医病理学研究；E-mail：385109522@qq.com
  • 基金资助:
    国家自然科学基金青年科学基金资助项目（81601646）；国家自然科学基金面上基金资助项目（81571852）

Combination of mRNA of Repair-related Genes in Rat Skeletal Muscles for Wound Age Estimation

LU Han-lin, DANG Li-hong, LI Na, DONG Ta-na, DU Qiu-xiang, WANG Ying-yuan, SUN Jun-hong   

  1. School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China
  • Online:2019-04-25 Published:2019-04-28

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摘要： 目的 探讨DNA聚合酶δ相互作用蛋白3（polymerase delta-interacting protein 3，POLDIP3）、类染色体浓缩调节样蛋白（regulator of chromosome condensation 1 like，RCC1L）、高脯氨酸蛋白5（proline-rich 5，PRR5）、核糖核酸输出蛋白1（ribonucleic acid export 1，RAE1） 4种修复相关基因的mRNA联合应用于早、中期损伤时间推断的方法。 方法 制备大鼠骨骼肌挫伤模型，于损伤后4、8、12、16、20、24、28、32、36、40、44和48 h取挫伤区肌肉组织，观察大鼠骨骼肌挫伤后修复过程中的组织形态学变化。利用逆转录实时定量聚合酶链反应（reverse transcription real-time quantitative polymerase chain reaction，RT-qPCR）检测Poldip3、Rcc1l、Prr5、Rae1 mRNA的相对表达量。利用4种基因在损伤后各时间点的表达模式对损伤过程进行分段，再通过Fisher判别法对上述分段结果进行准确性验证。 结果 组织形态学变化结果显示，骨骼肌挫伤后随着时间的延长发生修复，联合4种基因的表达趋势可将48 h内的损伤时间分为4~12 h、16~28 h、32~48 h 3个时间段，Fisher判别分析结果显示此3个时间段分类的准确率分别为83.3%、75.0%、73.3%。 结论 根据各基因相对表达量进行分组判别的准确度较高，联合多种基因mRNA的相对表达较单一指标进行损伤时间推断更为准确，结合Fisher判别分析可应用于早、中期损伤时间推断。

关键词: 法医病理学, 骨骼肌, 聚合酶&delta, 相互作用蛋白3, 类染色体浓缩调节样蛋白, 高脯氨酸蛋白5, 核糖核酸输出蛋白1, 损伤时间推断, 大鼠

Abstract: Objective To investigate the estimation of early and mid-term wound age by a combination of four mRNAs, the DNA polymerase delta-interacting protein 3 （POLDIP3） mRNA, regulator of chromosome condensation 1 like （RCC1L） mRNA, proline-rich 5 （PRR5） mRNA, and ribonucleic acid export 1 （RAE1） mRNA in rats skeletal muscles. Methods The model of rat skeletal muscle contusion was established, and then contusion area muscle tissue was extracted 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44 and 48 h after injury. Histomorphological changes during the repair process after rat skeletal muscle contusion were observed. The relative expressions of Poldip3, Rcc1l, Prr5 and Rae1 mRNAs were detected by reverse transcription real-time quantitative polymerase chain reaction （RT-qPCR）. Different stages of wound age were classified by using the expression patterns of four genes at various time points after injury. The accuracy of the results was verified by Fisher discriminant analysis. Results Histomorphological results showed that the repair process after skeletal muscle contusion occurred with the prolonging of time. Through combination of the expression trends of the four kinds of mRNAs, the 48 h after injury could be divided into three periods, 4-12 h, 16-28 h and 32-48 h. The Fisher discrimination method showed that the classification accuracy rates of the three periods were 83.3%, 75.0% and 73.3%, respectively. Conclusion The classification discrimination based on the relative expression of every gene has a higher accuracy, and the accuracy of wound age estimation with combination of mRNA relative expressions is higher than that with a single indicator. By combining with Fisher discrimination method, this method can be used for early and mid-term wound age estimation.

Key words: forensic pathology, skeletal muscle, polymerase delta-interacting protein 3, regulator of chromosome condensation 1 like, proline rich 5, ribonucleic acid export 1, wound age estimation, rats