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小鼠皮肤伤口愈合过程中促炎细胞因子mRNA的表达:法医学伤口年龄估计的初步研究(II)

The expression of mRNA of proinflammatory cytokines during skin wound healing in mice: a preliminary study for forensic wound age estimation (II).

作者信息

Sato Y, Ohshima T

机构信息

Department of Legal Medicine, Kanazawa University Faculty of Medicine, School of Medicine, Japan.

出版信息

Int J Legal Med. 2000;113(3):140-5. doi: 10.1007/s004140050285.

Abstract

This study examined the temporal expression of mRNA for the proinflammatory cytokines, interleukin (IL)-1alpha, IL-1beta, IL-6 and tumor necrosis factor-alpha (TNFalpha) in incised wounds in mice using the reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) techniques. After incision, an increase in each cytokine mRNA level was observed by RT-PCR. The local IL-6 mRNA level peaked at 6 h, while the peak levels of mRNA for IL-1alpha, IL-1beta and TNFalpha occurred between 48 and 72 h. All cytokine mRNA levels were almost normalized after 240 h. In the early phase of wound healing, infiltrating polymorphonuclear cells were labeled with antisense probes for IL-1alpha, IL-1beta and TNFalpha mRNA by the ISH technique. Thereafter, infiltrating mononuclear cells and spindle-shaped mesenchymal cells showed positive signals for all the cytokines examined. Regenerating epidermal cells were also labeled with the antisense probes for IL-1alpha, IL-6 and TNFalpha mRNA, indicating that IL-1, IL-6, and TNFalpha are involved in skin wound healing and their local production by various cells involved in the healing process is suggested. From the viewpoint of forensic pathology, the temporal characteristics of the cytokine mRNA expression may have a potential to indicate wound age or wound vitality.

摘要

本研究采用逆转录 - 聚合酶链反应(RT-PCR)和原位杂交(ISH)技术,检测了小鼠切割伤口中促炎细胞因子白细胞介素(IL)-1α、IL-1β、IL-6和肿瘤坏死因子-α(TNFα)的mRNA的时间表达情况。切割后,通过RT-PCR观察到每种细胞因子mRNA水平均升高。局部IL-6 mRNA水平在6小时达到峰值,而IL-1α、IL-1β和TNFα的mRNA峰值水平出现在48至72小时之间。240小时后,所有细胞因子mRNA水平几乎恢复正常。在伤口愈合的早期阶段,通过ISH技术,浸润的多形核细胞用针对IL-1α、IL-1β和TNFα mRNA的反义探针进行标记。此后,浸润的单核细胞和梭形间充质细胞对所有检测的细胞因子均显示阳性信号。再生的表皮细胞也用针对IL-1α、IL-6和TNFα mRNA的反义探针进行标记,这表明IL-1、IL-6和TNFα参与皮肤伤口愈合,并且提示它们由愈合过程中涉及的各种细胞进行局部产生。从法医病理学的角度来看,细胞因子mRNA表达的时间特征可能有潜力指示伤口的年龄或伤口的活力。

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