法医学杂志

法医学杂志 ›› 2020, Vol. 36 ›› Issue (3): 293-298.DOI: 10.12116/j.issn.1004-5619.2020.03.001

• 论著 •    下一篇

大鼠生前与死后损伤肌肉组织中自噬相关蛋白表达

时福霞， 李鹏， 鲁翰霖， 李娜， 杜秋香， 王英元， 孙俊红   

  1. 山西医科大学法医学院，山西 晋中 030600
  • 发布日期:2020-06-25 出版日期:2020-06-28
  • 通讯作者: 孙俊红，男，教授，主要从事损伤病理学和猝死病理学研究；E-mail：junhong.sun@sxmu.edu.cn
  • 作者简介:时福霞（1990—），女，硕士研究生，主要从事法医病理学研究；E-mail：fuxiashi@126.com
  • 基金资助:
    国家自然科学基金面上基金资助项目（81571852）；山西省高等学校大学生创新创业训练计划资助项目（2019191）

Expression of Autophagy-Associated Protein in Rat Muscle Tissues after Antemortem and Postmortem Injury

SHI Fu-xia, LI Peng, LU Han-lin, LI Na, DU Qiu-xiang, WANG Ying-yuan, SUN Jun-hong   

  1. School of Forensic Medicine, Shanxi Medical University, Jinzhong 030600, Shanxi Province, China
  • Online:2020-06-25 Published:2020-06-28

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摘要： 目的 研究大鼠骨骼肌损伤后BECN1、LC3和p62三种自噬相关蛋白的表达，探讨其在鉴别生前与死后损伤中的应用。 方法 72只健康成年Sprague-Dawley大鼠被随机分为未损伤的对照组、生前损伤组（0.5、1、2、4、8、16和24 h）和死后损伤组（0.5、1、2和4 h），制作大鼠右后肢损伤模型。运用Western印迹法检测对照组、生前损伤组以及死后损伤组骨骼肌中自噬蛋白BECN1、LC3-2/LC3-1和p62的表达，分别将数据中心化和标准化处理，建立生前损伤和死后损伤正交偏最小二乘判别分析（orthogonal partial least square-discriminate analysis，OPLS-DA）鉴别模型。 结果 大鼠骨骼肌挫伤后BECN1、p62蛋白的表达以及LC3-2/LC3-1值在生前损伤和死后损伤组中虽有不同程度的变化，但差异无统计学意义。BECN1的表达量及LC3-2/LC3-1值经中心化和标准化处理后，可区分生前损伤和死后损伤。生前损伤和死后损伤OPLS-DA模型提取的主成分对模型的解释程度较好（Rx2=0.563，Ry2=0.439），但预测程度稍差（Q2=0.366）。 结论 大鼠骨骼肌损伤局部组织中BECN1、LC3-2/LC3-1值可用于生前损伤和死后损伤的鉴别。

关键词: 法医病理学, 创伤和损伤, 自噬相关蛋白, 骨骼肌, 生前损伤, 死后损伤, 大鼠

Abstract: Objective To study the expression of the three autophagy-associated proteins, BECN1, LC3 and p62, after the injury of the skeletal muscle of rats and to explore its application in differentiation between antemortem and postmortem injury. Methods The 72 healthy Sprague-Dawley rats were randomly divided into the undamaged control group, the antemortem injury group （0.5 h, 1 h, 2 h, 4 h, 8 h, 16 h and 24 h） and postmortem injury group （0.5 h, 1 h, 2 h and 4 h）. A model of the injured right hind limb of rats was constructed. The expressions of the autophagy-associated proteins, BECN1, LC3-2/LC3-1 and p62, in the control group, the antemortem injury group and postmortem injury group were detected by Western blotting method. The data were respectively centralized and standardized and the orthogonal partial least square-discrimination analysis （OPLS-DA） identification model of antemortem and postmortem injury groups was constructed. Results The expression of BECN1, p62 protein and LC3-2/LC3-1 after the injury of the skeletal muscle of the rats showed different degrees of changes, but the differences among the 3 groups had no statistical significance. Antemortem and postmortem injury groups can be distinguished by centralizing and standardizing the expression levels of autophagy protein BECN1 and the ratio of LC3-2/LC3-1. The principal components extracted from OPLS-DA model of antemortem injury and postmortem injury had a relatively good interpretation of the model （Rx2=0.563, Ry2=0.439）, but it were less predictive （Q2=0.366）. Conclusion The expression of BECN1 and the ratio of LC3-2/LC3-1 in injured local tissue of the rat skeletal muscle can be used for the differentiation of antemortem injury group and postmortem injury group.

Key words: forensic pathology, wounds and injuries, autophagy-associated protein, skeletal muscle, antemortem injury, postmortem injury, rats