【摘要】： 目的:應(yīng)用熒光原位雜交(FISH)技術(shù)檢測大鼠皮膚和骨骼肌挫傷后皮膚、肌肉組織中細(xì)胞間黏附因子-1(ICAM-1)mRNA的時序性表達(dá)規(guī)律,探討這種基因mRNA表達(dá)在損傷時間推斷中的可行性,旨為法醫(yī)實踐中推斷早期皮膚和肌肉損傷時間提供科學(xué)的實驗依據(jù)。 方法:選取健康成年SD大鼠48只,隨機(jī)分為對照組(6只)和實驗組(42只)。均實施麻醉、褪毛后,實驗組大鼠采用自由落體打擊法于右后肢股四頭肌部位造成皮膚、肌肉挫傷模型,損傷后0.5h、1h、6h、12h、18h、24h、30h七個時間點(每個時間點6只)取挫傷處皮膚及肌肉組織各30mg,置于液氮中保存?zhèn)溆?對照組大鼠未打擊造傷,其余處理與實驗組相同。檢材冰凍切片后進(jìn)行熒光原位雜交,激光共聚焦顯微鏡觀察、拍照,采用Image-pro plus進(jìn)行圖像分析。對照組與各損傷組數(shù)據(jù)的處理采用SSPS10.0統(tǒng)計軟件進(jìn)行方差分析和SNK檢驗。 結(jié)果:大鼠皮膚和骨骼肌挫傷后ICAM-1 mRNA在皮膚組織中的表達(dá)0.5h達(dá)到峰值,24h下降至正常對照組的2.25倍隨后繼續(xù)升高;在肌肉組織中于損傷后0.5h表達(dá)顯著增強(qiáng),6h達(dá)到峰值,18h降至最低后再次升高。 結(jié)論:大鼠皮膚、骨骼肌挫傷后皮膚和肌肉組織中ICAM-1 mRNA表達(dá)隨著損傷時間呈規(guī)律性變化,可望用于法醫(yī)學(xué)鑒定及早期法醫(yī)學(xué)損傷時間推斷。熒光原位雜交技術(shù)是一種分子細(xì)胞遺傳學(xué)技術(shù),敏感而且準(zhǔn)確,適合法醫(yī)學(xué)研究及檢案的需要。
[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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：D919

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