發(fā)布時(shí)間：2018-11-06 20:25

【摘要】： 目的:建立體外培養(yǎng)神經(jīng)細(xì)胞和大鼠創(chuàng)傷性腦損傷動(dòng)物模型,研究損傷后培養(yǎng)神經(jīng)細(xì)胞形態(tài)學(xué)改變和大鼠海馬蛋白質(zhì)組表達(dá)情況,尋找差異性生物標(biāo)志蛋白。 方法:選新生24小時(shí)內(nèi)SD大鼠,取海馬組織,培養(yǎng)10-14天后,隨機(jī)分為對(duì)照組及損傷后4h、8h、12h、24h和48h組,復(fù)制Scott's神經(jīng)細(xì)胞液壓沖擊損傷模型,進(jìn)行神經(jīng)細(xì)胞形態(tài)學(xué)觀察。選成年雄性SD大鼠隨機(jī)分為手術(shù)對(duì)照組及損傷后4h、8h、12h、24h和48h組,復(fù)制Marmarou's落體打擊大鼠腦損傷模型,采用雙向凝膠電泳技術(shù)分析大鼠閉合性腦損傷后不同時(shí)間海馬中蛋白質(zhì)組表達(dá)的改變。 結(jié)果:1體外培養(yǎng)神經(jīng)細(xì)胞液壓沖擊損傷裝置穩(wěn)定性評(píng)價(jià):通過(guò)記錄小室內(nèi)和有機(jī)玻璃管內(nèi)的壓力大小來(lái)檢測(cè)液壓沖擊損傷壓力的大小,結(jié)果表明,小室內(nèi)和有機(jī)管內(nèi)的壓力存在一線性關(guān)系(Y=4.4763X+0.2986,Y代表有機(jī)玻璃管內(nèi)壓力,X代表小室內(nèi)壓力),造成細(xì)胞損傷力的大小以小室內(nèi)的壓力為準(zhǔn)。有機(jī)玻璃管和小室內(nèi)壓力隨鐘擺打擊角度的增加而增大。輕(0.1MPa)、中(0.2MPa)和重度(0.3MPa)液壓沖擊損傷所需鐘擺下落角度分別為13.5°、21.5°和27°。2液壓沖擊損傷神經(jīng)細(xì)胞形態(tài)學(xué)改變:液壓沖擊后,神經(jīng)細(xì)胞甲苯胺藍(lán)染色,正常神經(jīng)細(xì)胞胞核呈藍(lán)色,胞漿染成淡藍(lán)色,損傷組培養(yǎng)神經(jīng)細(xì)胞主要表現(xiàn)為部分神經(jīng)細(xì)胞胞體濃縮,胞核固縮,染色較深,呈深藍(lán)色,多為壞死型損傷細(xì)胞,
[Abstract]:Aim: to establish an animal model of cultured nerve cells and traumatic brain injury (TBI) in vitro, to study the morphological changes of cultured neurons and the proteome expression in hippocampus of rats after injury, and to search for different biomarkers. Methods: SD rats within 24 hours were selected and cultured for 10-14 days. The rats were randomly divided into two groups: the control group and the control group. The Scott's nerve cells were induced by hydraulic shock for 24 hours and 48 hours after the injury. The morphology of nerve cells was observed. Adult male SD rats were randomly divided into two groups: the control group and the control group. The rats were divided into two groups at 4 h, 8 h, 12 h and 48 h after injury. The model of brain injury was induced by Marmarou's falling body. Two-dimensional gel electrophoresis was used to analyze the changes of proteome expression in hippocampus of rats after closed brain injury at different time points. Results: 1 Stability evaluation of the hydraulic shock injury device for cultured nerve cells in vitro: the hydraulic impact injury pressure was measured by recording the pressure in the chamber and in the plexiglass tube. There was a linear relationship between the pressure in the chamber and the pressure in the organic tube (YY 4.4763X 0.2986 Y for the pressure in the plexiglass tube and X for the pressure in the chamber), and the damage force of the cells was determined by the pressure of the chamber. The pressure of plexiglass tube and chamber increases with the increase of pendulum angle. The falling angles of pendulum for 0.1MPa, 0.2MPa and 0.3MPa were 13.5 擄, 21.5 擄and 27 擄respectively. The nerve cells were stained with toluidine blue, the nuclei of normal neurons were blue and the cytoplasm was light blue. In the injured group, the nerve cells in the injured group were mainly concentrated in the cell bodies, pyknosis of the nuclei, deep staining and dark blue. Most of them are necrotic injury cells.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R363

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