大鼠骨骼肌损伤修复中FZD4时序性变化及其应用 |
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引用本文: | 孙俊红,朱细燕,张小红,刘起清,杜秋香,王英元. 大鼠骨骼肌损伤修复中FZD4时序性变化及其应用[J]. 中国法医学杂志, 2016, 0(5): 444-447. DOI: 10.13618/j.issn.1001-5728.2016.05.004 |
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作者姓名: | 孙俊红 朱细燕 张小红 刘起清 杜秋香 王英元 |
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作者单位: | 1. 山西医科大学法医学院,山西太原,030001;2. 山西医科大学法医学院,山西太原030001; 邹平县公安局,山东滨州 256200 |
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基金项目: | 国家自然科学基金委青年基金资助项目,山西省回国留学人员科研资助项目 |
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摘 要: | 目的以卷曲受体4(FZD4)为例,探索细胞膜上受体蛋白的表达变化能否作为损伤时间推断的指标。方法 78只大鼠随机分为对照组和损伤后4h、8h、12h、16h、20h、24h、28h、32h、36h、40h、44h和48 h组(n=6)。麻醉后,采用重力锤自由落体方式砸伤大鼠右后肢,取股四头肌处挫伤的组织。应用免疫荧光、real-time PCR和western blotting方法检测肌肉中FZD4蛋白及m RNA的表达量。结果免疫荧光结果显示FZD4表达于骨骼肌细胞膜上,胞浆和细胞核内均未见表达;Real-time PCR结果表明FZD4 m RNA在损伤后8h、12h、36h和40h显著上调(P0.05),均超过对照组的两倍;Western blotting结果示FZD4蛋白的表达在8h、36h、40h、44h和48h组明显增加(P0.05),但均未超过对照组的两倍。结论 FZD4 m RNA和蛋白在肌肉挫伤后呈时序性变化规律可作为损伤时间推断的指标,在一定程度上FZD4 m RNA比FZD4更适用于损伤时间推断。
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关 键 词: | 法医病理学 损伤时间推断 卷曲受体4 Wnt信号通路 RT-PCR western blotting 免疫荧光 |
Time-dependent expression of FZD4 in contused skeletal muscle of rat used for wound age estimation |
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Abstract: | Objective Take frizzled class receptor 4(FZD4) as an example for exploring whether the transmembrane receptor protein is a suitable marker for wound age estimation. Methods A total of 78 Sprague-Dawley rats were divided randomly into the control group and the contusion groups of 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, and 48h after injury (n=6). Using a drop-ball technique, a contusion was produced in the right lamb of rats. The samples were dissected from injury zones. Then the expression of FZD4 was detected using immunofluorescence, real-time PCR and western blot, respectively. Results FZD4 was located on the membrane of skeletal muscle cells. Compared to the control group, FZD4 mRNA showed a signiifcant increase (over 2-fold) in 8h, 12h, 36h, and 40h after injury by Real-time PCR (P<0.05), and FZD4 protein showed a statistical up-regulation (less than 2-fold) in 8h, 36h, 40h, 44h, and 48h post contusion by western blotting (P<0.05). Conclusion The expression of FZD4 mRNA and protein are both time-dependent during contused skeletal muscle repair. To some degree, FZD4 mRNA was more suitable than corresponding protein for determining wound age. |
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Keywords: | forensic pathology wound age estimation frizzled class receptor 4 Wnt signaling pathway real-time PCR western blot immunolfuorescence |
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