title: Simultaneous time course analysis of multiple markers based on DNA microarray in incised wound in skeletal muscle for wound aging.
abstract: Assessment of incised wound age in skeletal muscles is important because fatal injuries are often complicated with muscle involvement. 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 injury 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 48h post-injury. DNA microarray analysis using RNA extracted from the muscle samples of 12h post-injury identified 3,655 upregulated and 3,583 downregulated 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-1β), 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 expression from 6 through 36h post injury. Upregulation of CCL4 and IL-1β was also persisted until 36h post injury. IL-6 mRNA was highly and rapidly expressed at 6h post-injury followed by significant decrease at 12h. Unlike other four cytokines, IL-7 showed slow and steady increasing over time until 48h 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.
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan. Electronic address: drgaba@med.nagoya-cu.ac.jp.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Hassan Gaballah
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Mamiko
authors.initials: M
authors.lastname: Fukuta
authors.affiliation: Department of Food and Nutritional Sciences, Chubu University College of Bioscience and Biotechnology 1200 Matumoto-cho Kasugai, 487-8501, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Maeno
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Seko-Nakamura
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Monma-Ohtaki
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Shibata
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Kato
authors.affiliation: Department of Forensic Medicine, Nagoya City University Graduate School of Medical Sciences 1 Kawasumi Mizuho-cho Mizuho-ku Nagoya, 467-8601, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Aoki
authors.affiliation: Department of Legal Medicine, Iwate Medical University 2-1-1 Nishitokuta, Yahaba-cho Shiwagun, Iwate 028-3694, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Takamiya
mesh_terms: D000818:Animals; D016207:Cytokines / Q000235:genetics; D049429:Forensic Pathology / Q000379:methods*; D005786:Gene Expression Regulation; D015850:Interleukin-6 / Q000235:genetics; D051379:Mice; D018482:Muscle, Skeletal / Q000293:injuries* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D013997:Time Factors; D014945:Wound Healing / Q000235:genetics*
chemical_list: D016207:Cytokines; D015850:Interleukin-6
keywords: Cytokines; DNA microarray; Gene expression; Incised wound; Skeletal muscle; Wound aging