Nagoya City University Academic Repository
学 位 の 種 類 博士 (医学)
報 告 番 号 甲第１５５４号
学 位 記 番 号 第１１０９号
氏 名 Mohammed Hassan Gaballah
授 与 年 月 日 平成 29 年 3 月 24 日
学位論文の題名
Simultaneous time course analysis of multiple markers based on DNA microarray in incised wound in skeletal muscle for wound aging (骨格筋切創の受傷後経過時間推定のための DNA マイクロアレイに基づく 各種マーカーの経時的動態の同時解析)
Forensic Science International 2016; 266: 357-368
論文審査担当者 主査： 大塚 隆信
Simultaneous time course analysis of multiple markers based on
DNA microarray in incised wound in skeletal muscle for wound
aging
Mohammed Hassan Gaballah Abstract
The determination of wound age is essential in forensic practice. A vari-ety of methods for wound age estimation have been investigated and these methods were reviewed by forensic pathologists including routine histopatho-logical examination, immunohistochemical staining and reverse transcription PCR (RT-PCR). Recently, molecular pathology techniques have improved the diagnostic abilities of forensic pathologists in various fields including determination cause of death, and estimation of postmortem interval and wound age. In this study, transcriptome of injured skeletal muscle along with histopathological and immunohistochemistry staining, were analyzed to explore the biological effect of incised injuries using a mouse incised in-jury model. An incisional wound was made at the biceps femoris muscle of anesthetized mice, and the muscles were sampled at 6, 12, 24, 36 and 48 h post-injury. DNA microarray analysis using RNA extracted from the muscle samples of 12 h post-injury identified 3,655 upregulated and 3,583 downregu-lated genes. Referring to the results of the gene ontology and gene expression pathway analysis, time course expression of five cytokines, namely chemokine (C-C motif) ligand 4 (CCL4), chemokine (C-X-C motif) ligand 5 (CXCL5), interleukin-1 beta (IL-1b),interleukin- 6 (IL-6) and interleukin-7 (IL-7), were analyzed by quantative reverse transcription PCR (qRT-PCR). CXCL5 was the most upregulated gene throughout the post-injury period with higher ex-pression from 6 through 36 h post injury. Upregulation of CCL4 and IL-1b was also persisted until 36 h post injury. IL-6 mRNA was highly and rapidly expressed at 6 h post-injury followed by significant decrease at 12 h. Un-like other four cytokines, IL-7 showed slow and steady increasing over time until 48 h post-injury. Immunohistochemical staining of post-injury samples showed gradual mild increase of staining intensity proportional to increasing time points especially around the wound edges. The present study highlights the unique dynamics of each cytokine and reflects their roles in the process of muscle wound healing, and suggests the potential of them as a tool for forensic wound age estimation.
