Time-dependent expression of SNAT2 mRNA in the contused skeletal muscle of rats: a possible marker for wound age estimat
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ORIGINAL ARTICLE
Time-dependent expression of SNAT2 mRNA in the contused skeletal muscle of rats: a possible marker for wound age estimation Qiu-xiang Du • Jun-hong Sun • Ling-yu Zhang Xin-hua Liang • Xiang-jie Guo • Cai-rong Gao Ying-yuan Wang
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Accepted: 13 August 2013 / Published online: 18 September 2013 Ó Springer Science+Business Media New York 2013
Abstract To estimate the age of skeletal muscle contusion, the expression of SNAT2 mRNA in contused skeletal muscle of rats was detected by real-time polymerase chain reaction (PCR). In total, 78 Sprague–Dawley male rats were divided into control and contusion groups. At 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, and 48 h (n = 6) after contusion, the rats were sacrificed with a lethal dose of pentobarbital. Another 24 rats received contusion injuries at 6, 12, 18, and 24 h (n = 6) after death. Total RNA was isolated from muscle specimens using the TRIzol reagent and reverse-transcribed into first-strand cDNA. Sequencespecific primers and TaqMan fluorogenic probes for SNAT2 mRNA and RPL13 mRNA were designed using the AlleleID 6 software, and the expression levels of SNAT2 mRNA were determined by real-time PCR. At 4, 16, 20, and 24 h after contusion, expression levels of SNAT2 mRNA normalized to RPL13 mRNA increased by 2.07 (P \ 0.05), 2.53 (P \ 0.05), 2.68 (P \ 0.05), and 2.06 fold (P \ 0.05) respectively, versus that in the control group. However, there was no significant change in the expression level of SNAT2 mRNA from 24 to 48 h (P [ 0.05) after contusion, when normalized to RPL13 mRNA. There was no change in the expression level of SNAT2 mRNA between the normal skeletal muscle from the left limb of the same injured rat and the control. Also, no degradation of SNAT2 mRNA was detected in the postmortem samples (P [ 0.05). This result suggests that
Q. Du J. Sun (&) L. Zhang X. Liang X. Guo C. Gao Y. Wang Department of Forensic Pathology, Shanxi Medical University, 56 South Xinjian Nan Road, Taiyuan 030001, Shanxi, People’s Republic of China e-mail: [email protected]
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the determination of SNAT2 mRNA levels by real-time PCR may be useful for estimating wound age. Keywords Wound age estimation Contused skeletal muscle SNAT2 mRNA Real-time PCR
Introduction Historically, wound age estimation has been one of the most common areas of forensic research. With the development of genetically engineered animals molecules essential for wound healing have been identified; in particular, cytokines and growth factors are useful markers for the determination of wound vitality or age [1]. However, wound age estimation remains an unsolved challenge for forensic pathologists. It is important to identify more suitable markers for determining wound vitality or age in forensic medicine. Wound healing is a complex series of reactions and interactions among cells and ‘‘mediators.’’ As a general rule, the processes are divided into three phases: inflammation, proliferation, and maturation [1]. Damage to skeletal muscle due to blunt trauma presents patholog
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