Comparison of the homogeneity of mRNAs encoding SFRP5, FZD4, and Fosl1 in post-injury intervals: Subcellular localization of markers may influence wound age estimation
Comparison of the homogeneity of mRNAs encoding SFRP5, FZD4, and Fosl1 in post-injury intervals: Subcellular localization of markers may influence wound age estimation
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损伤后时间间隔内编码 SFRP5、FZD4 和 Fosl1 的 mRNA 的同质性比较：标记物的亚细胞定位可能影响伤口年龄估计
DOI：
10.1016/j.jflm.2016.07.013
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发表时间：
2016
影响因子：
1.5
通讯作者：
Sun Jun-hong
中科院分区：
文献类型：
--
作者：
Zhu Xi-yan;Du Qiu-xiang;Li San-qiang;Sun Jun-hong
在法医学中，利用mRNA准确判断伤口年龄时，必须考虑组间和组内相对表达的异质性。本研究的目的是评估损伤后各组SFRP5、FZD4和Fosl1 mrna的同质性。相应的蛋白显示不同的亚细胞位置。78只Sprague-Dawley大鼠分为对照组和挫伤组。在挫伤后4、8、12、16、20、24、28、32、36、40、44或48小时（每组n = 6）（腹腔水合氯醛麻醉），用致死剂量戊巴比妥处死大鼠，收集损伤肌肉样本。采用real-time PCR方法获取SFRP5、FZD4和Fosl1 mrna的原始Ct值。归一化为RPL13 mRNA水平后，计算各归一化Ct的变异系数（CV）、相对平均偏差（d%）及其在损伤后各时间间隔的相对表达水平。采用两种方法比较三个基因的同质性。首先，根据每个损伤后间隔的CV值对每个基因进行评分。然后，计算13个分数之和；低的总和表示高的同质性。其次，将计算得到的13个cv或d%s作为原始数据，用mean±SD表示。基于该平均值±SD，计算CV的CV和d%s的d%来表示变异；低值表明同质性高。FZD4 mRNA的变异总和低于SFRP5和Fosl1 mRNA，这与FZD4 mRNA在三个基因中具有最低的平均值、最小的CV和最小的d%s的结果一致。综上所述，这些数据表明，在损伤后的时间间隔内，编码膜质FZD4的mRNA可能比编码SFRP5和Fosl1的mRNA更均匀。
The inter-group heterogeneity and intra-group homogeneity of relative expression are very necessary when the mRNA were used to determine wound age accurately in forensic medicine. The aim of this study was to assess the intra-group homogeneity of SFRP5, FZD4 and Fosl1 mRNAs in post-injury intervals. The corresponding proteins show different subcellular locations. A total of 78 Sprague–Dawley rats were divided into control and contusion groups. At 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, or 48 h (n = 6 per group) after contusion (under anesthesia by chloral hydrate intraperitoneally), the rats were sacrificed using a lethal dose of pentobarbital, and samples of the injured muscles were collected. The raw Ct values of SFRP5, FZD4, and Fosl1 mRNAs were obtained using real-time PCR. After normalized to RPL13 mRNA levels, the coefficient of variation (CV) and the relative average deviation (d%) of each normalized Ct, and their relative expression levels, were calculated in each post-injury interval. Two methods were applied to compare the homogeneity of the three genes. First, each gene was given a score based on its CV value in each post-injury interval. Then, the sum of the 13 scores was calculated; a low sum indicated high homogeneity. Second, the 13 calculated CVs or d%s were used as raw data, which was described as the mean ± SD. Based on this mean ± SD, a CV of the CVs and a d% of the d%s were calculated to represent the variation; a low value indicated high homogeneity. The sum of the variability of FZD4 mRNA was lower than those of the SFRP5 and Fosl1 mRNAs, consistent with the results that the FZD4 mRNA had the lowest mean, the smallest CV of all CVs, and the smallest d% of all d%s, among the three genes. In conclusion, these data indicated that mRNA encoding membranous FZD4 was likely to be more homogeneous than those encoding SFRP5 and Fosl1 within post-injury intervals.