失血性休克大鼠海馬中HSP70的表達(dá)及死后穩(wěn)定性研究
發(fā)布時(shí)間:2018-06-23 21:02
本文選題:失血性休克 + 熱休克蛋白 ; 參考:《重慶醫(yī)科大學(xué)》2008年碩士論文
【摘要】: 背景及目的:失血性休克死亡是交通事故、醫(yī)療責(zé)任事故以及刑事犯罪中一個(gè)常見(jiàn)的死因。目前對(duì)于失血性休克的研究,多集中于失血性休克的補(bǔ)液治療及缺血-再灌注損傷的機(jī)制方面,而對(duì)失血性休克中出血至死亡所經(jīng)歷時(shí)間的推斷,則報(bào)道甚少。至今對(duì)失血性休克發(fā)生后傷者存活時(shí)間的推斷,仍沒(méi)有一個(gè)公認(rèn)的標(biāo)準(zhǔn),也成為法醫(yī)病理學(xué)上的一大難題。熱休克蛋白70(heat shock protein 70,HSP70)是機(jī)體遭受缺血、缺氧、創(chuàng)傷、感染等各種應(yīng)激性刺激后誘導(dǎo)HSP70基因的表達(dá)而產(chǎn)生的一種熱休克蛋白,它可在多種細(xì)胞中表達(dá),并呈現(xiàn)出一定的規(guī)律性。本實(shí)驗(yàn)主要研究大鼠失血性休克后海馬中HSP70蛋白的表達(dá)和其死后穩(wěn)定性,并探討其法醫(yī)學(xué)意義。 方法:1、選材:實(shí)驗(yàn)動(dòng)物來(lái)源于重慶醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心飼養(yǎng)的健康的清潔的SD大鼠105只,按照隨機(jī)的原則將大鼠分為正常組、假手術(shù)組、失血性休克組、創(chuàng)傷對(duì)照組和死后穩(wěn)定性組。失血性休克組與死后穩(wěn)定性組采用改良Wigger’s失血性休克模型,創(chuàng)傷對(duì)照組參照Feeney’S自由落體模型裝置制作大鼠腦挫裂傷模型;2、取材:在失血性休克/創(chuàng)傷后不同時(shí)間點(diǎn)取大鼠海馬組織;3、對(duì)取材海馬組織石蠟包埋切片進(jìn)行HE染色光鏡觀察;4、采用免疫組織化學(xué)染色法進(jìn)行HSP70染色;5、使用SPSS 10.0軟件包對(duì)所得數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果:失血性休克1小時(shí)后,海馬組織和血管壁即可見(jiàn)HSP70陽(yáng)性表達(dá),此后表達(dá)逐漸增強(qiáng),至2小時(shí)呈強(qiáng)陽(yáng)性表達(dá)(P0.01),部分錐體細(xì)胞核內(nèi)也可見(jiàn)陽(yáng)性表達(dá)。創(chuàng)傷對(duì)照組僅在血管壁和紅細(xì)胞見(jiàn)HSP70陽(yáng)性表達(dá),海馬組織HSP70表達(dá)呈弱陽(yáng)性。死后穩(wěn)定性組表達(dá)情況與休克2小時(shí)組相似(P0.05)。 結(jié)論:大鼠失血性休克后海馬中HSP70蛋白的表達(dá)具有一定的規(guī)律性,呈現(xiàn)出逐漸升高-高峰期的變化,其表達(dá)水平在傷后2小時(shí)達(dá)到高峰,且在-20℃條件下,死后穩(wěn)定性好。因此,HSP70蛋白可作為法醫(yī)學(xué)失血性休克后個(gè)體存活時(shí)間推斷的一個(gè)參考指標(biāo)。
[Abstract]:Background and objective: hemorrhagic shock death is a common cause of death in traffic accidents, medical accidents and criminal offences. At present, the researches on hemorrhagic shock mainly focus on the rehydration therapy of hemorrhagic shock and the mechanism of ischemia-reperfusion injury, but there are few reports on the inference of the time between bleeding and death in hemorrhagic shock. So far, the estimation of the survival time of the injured after hemorrhagic shock has not been a recognized standard, and has become a major problem in forensic pathology. Heat shock protein 70 (heat shock protein 70) is a heat shock protein (HSP70) which is induced by various stress stimuli, such as ischemia, hypoxia, trauma, infection and so on. It can be expressed in many kinds of cells and has a certain regularity. In this study, the expression and postmortem stability of HSP70 protein in hippocampus of rats with hemorrhagic shock were studied and its forensic significance was discussed. Methods: 105 healthy and clean SD rats were collected from the Animal experiment Center of Chongqing Medical University. According to the random principle, the rats were divided into normal group, sham operation group and hemorrhagic shock group. Trauma control group and postmortem stability group. The hemorrhagic shock group and the postmortem stability group were treated with modified Wiggerers hemorrhagic shock model. The traumatic control group made rat brain contusion and laceration model according to FeeneyeyS free-fall model device. The materials were taken from hippocampus tissue of rats at different time points after hemorrhagic shock / trauma, and paraffin embedded sections of hippocampus tissue were obtained. HSP70 staining was performed by immunohistochemical staining and statistical analysis was carried out by SPSS 10.0 software package. Results: one hour after hemorrhagic shock, HSP70 positive expression was observed in hippocampal tissue and vascular wall, and then increased gradually, and was strongly expressed at 2 hours (P0.01). The positive expression was also found in some pyramidal nuclei. In trauma control group, HSP70 expression was only found in vascular wall and erythrocytes, and was weakly positive in hippocampal tissue. The expression of postmortem stability group was similar to that of shock group for 2 hours (P0.05). Conclusion: the expression of HSP70 protein in hippocampus of rats with hemorrhagic shock has a certain regularity, and the expression level reaches its peak at 2 hours after injury, and the postmortem stability is good at -20 鈩,
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