大鼠皮肤、骨骼肌挫伤后ICAM-1 mRNA表达与损伤时间的研究

发布时间：2019-05-30 02:22

【摘要】： 目的:应用荧光原位杂交(FISH)技术检测大鼠皮肤和骨骼肌挫伤后皮肤、肌肉组织中细胞间黏附因子-1(ICAM-1)mRNA的时序性表达规律,探讨这种基因mRNA表达在损伤时间推断中的可行性,旨为法医实践中推断早期皮肤和肌肉损伤时间提供科学的实验依据。方法:选取健康成年SD大鼠48只,随机分为对照组(6只)和实验组(42只)。均实施麻醉、褪毛后,实验组大鼠采用自由落体打击法于右后肢股四头肌部位造成皮肤、肌肉挫伤模型,损伤后0.5h、1h、6h、12h、18h、24h、30h七个时间点(每个时间点6只)取挫伤处皮肤及肌肉组织各30mg,置于液氮中保存备用;对照组大鼠未打击造伤,其余处理与实验组相同。检材冰冻切片后进行荧光原位杂交,激光共聚焦显微镜观察、拍照,采用Image-pro plus进行图像分析。对照组与各损伤组数据的处理采用SSPS10.0统计软件进行方差分析和SNK检验。结果:大鼠皮肤和骨骼肌挫伤后ICAM-1 mRNA在皮肤组织中的表达0.5h达到峰值,24h下降至正常对照组的2.25倍随后继续升高;在肌肉组织中于损伤后0.5h表达显著增强,6h达到峰值,18h降至最低后再次升高。结论:大鼠皮肤、骨骼肌挫伤后皮肤和肌肉组织中ICAM-1 mRNA表达随着损伤时间呈规律性变化,可望用于法医学鉴定及早期法医学损伤时间推断。荧光原位杂交技术是一种分子细胞遗传学技术,敏感而且准确,适合法医学研究及检案的需要。
[Abstract]:Objective: to detect the sequential expression of Intercellular Adhesion Factor-1 (ICAM-1) mRNA in rat skin and skeletal muscle after contusion by fluorescence in situ hybridization (FISH). To explore the feasibility of mRNA expression in the inference of injury time, in order to provide scientific experimental basis for inferred early skin and muscle injury time in forensic practice. Methods: 48 healthy adult SD rats were randomly divided into control group (n = 6) and experimental group (n = 42). All the rats in the experimental group were anesthetized. After hair removal, the model of skin and muscle contusion was established in the quadriceps femoris of the right hindlimb by free fall method. 0.5 h, 1 h, 6 h, 12 h, 18 h, 24 h, 1 h, 6 h, 18 h, 24 h, 0 h, 1 h, 6 h, 18 h, 24 h, respectively. 30 mg of skin and muscle tissue were taken at seven time points of 30 h (6 at each time point) and stored in liquid nitrogen for storage. The rats in the control group did not strike the injury, and the other treatments were the same as those in the experimental group. The frozen sections were stained with fluorescence in situ, observed by laser confocal microscope, photographed and analyzed by Image-pro plus. The data of control group and injury group were processed by SSPS10.0 statistical software for variance analysis and SNK test. Results: the expression of ICAM-1 mRNA in skin tissue reached the peak at 0.5 h after contusion of skin and skeletal muscle, and then decreased to 2.25 times of that in normal control group at 24 h and then continued to increase. In muscle tissue, the expression increased significantly at 0.5 h after injury, reached the peak at 6 h, decreased to the lowest at 18 h, and then increased again. Conclusion: the expression of ICAM-1 mRNA in skin and muscle tissue of rats after skin and skeletal muscle contusion changes regularly with the time of injury, which is expected to be used for forensic identification and inference of early forensic injury time. Fluorescence in situ hybridization is a kind of molecular cytogenetic technique, which is sensitive and accurate, and is suitable for forensic research and case detection.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

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