本文選題：去鐵胺　切入點(diǎn)：低氧誘導(dǎo)因子-1α　出處：《山西醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的：新生兒缺氧缺血性腦損傷(hypoxic-ischemic brain damage ,HIBD)是指各種圍生期窒息而導(dǎo)致腦的缺氧缺血性損害，臨床出現(xiàn)一系列腦病的表現(xiàn),是我國(guó)傷殘兒童致殘重要的原因之一。本實(shí)驗(yàn)通過(guò)建立新生大鼠缺氧缺血性腦損傷(HIBD)模型，觀察缺氧誘導(dǎo)因子1α(hypoxia-inducible factor 1α)表達(dá)與去鐵胺（deferoxamine，DFO）可能的保護(hù)機(jī)制，為其在新生兒缺氧缺血性腦病（hypoxic-ischemic encephalopathy, HIE）的臨床應(yīng)用提供理論依據(jù)。 方法：新生7日齡Wistar大鼠120只隨機(jī)分為假手術(shù)組（8只）、HIBD模型組及去鐵胺（DFO）組，后兩組再根據(jù)處死時(shí)間不同又分為7個(gè)亞組：3h組、6h組、12h組、24h組、48h組、3d組及7d組，每亞組各8只。假手術(shù)組給予頸正中切口游離左側(cè)頸總動(dòng)脈，不進(jìn)行缺血缺氧。模型采用阻斷左側(cè)頸總動(dòng)脈后置于含8%O2的低氧環(huán)境中2小時(shí)制備而成。去鐵胺組,制成HIBD動(dòng)物模型前24 h以去鐵胺(200mg/kg,溶于生理鹽水)單次腹腔內(nèi)注射。各組分別于不同時(shí)間點(diǎn)處死動(dòng)物,肉眼觀察腦組織大體形態(tài)變化，，光鏡下觀察各組大鼠左側(cè)腦組織HE染色的病理改變，并采用免疫組織化學(xué)方法檢測(cè)HIF-1α和Caspase3的表達(dá)。 結(jié)果：(1)缺氧缺血后新生大鼠出現(xiàn)不同程度的行為異常。(2)缺氧缺血后各時(shí)間點(diǎn)可見(jiàn)腦組織大體有不同程度的異常改變,去鐵胺干預(yù)后上述改變減輕。(3)HE染色：假手術(shù)組腦組織結(jié)構(gòu)及細(xì)胞層次清晰，神經(jīng)元排列整齊緊密；HIBD模型組各時(shí)間點(diǎn)出現(xiàn)不同程度的神經(jīng)細(xì)胞腫脹、變性、壞死，細(xì)胞核碎裂、溶解，神經(jīng)元數(shù)目減少，膠質(zhì)細(xì)胞增生。去鐵胺組各時(shí)間點(diǎn)損傷程度較HIBD組明顯減輕，神經(jīng)細(xì)胞存活數(shù)量多，細(xì)胞排列尚規(guī)則，僅見(jiàn)少量神經(jīng)細(xì)胞變性壞死。(4)免疫組織化學(xué)染色：HIF-1α及Caspase-3的陽(yáng)性表達(dá)均呈棕黃色細(xì)顆粒沉積，陽(yáng)性細(xì)胞可表達(dá)于大腦皮層和海馬，陽(yáng)性著色主要位于神經(jīng)細(xì)胞胞漿及突起。①HIF-1α的表達(dá)：假手術(shù)組HIF-1α呈低水平表達(dá)，各時(shí)間點(diǎn)無(wú)明顯變化；HIBD模型組各時(shí)間點(diǎn)均可見(jiàn)HIF-1α陽(yáng)性表達(dá)，同一時(shí)間點(diǎn)與假手術(shù)組相比，HIF-1α表達(dá)明顯增加，差異均有統(tǒng)計(jì)學(xué)意義（P0.05）。HIF-1α在HIBD后3h即增強(qiáng)，12h達(dá)高峰，之后逐漸降低；而DFO組則6h達(dá)高峰，12 h和24 h仍在較高水平，各時(shí)間點(diǎn)HIF-1α表達(dá)水平較HIBD組明顯升高,差異有統(tǒng)計(jì)學(xué)意義（P0.05）。②Caspase-3的表達(dá)：假手術(shù)組Caspase-3呈低水平表達(dá)，各時(shí)間點(diǎn)無(wú)明顯變化；HIBD模型組Caspase-3陽(yáng)性表達(dá)，6h明顯升高, 12h和24h仍在較高水平，7d開始降低仍保持一定的表達(dá)水平，且各時(shí)間點(diǎn)表達(dá)高于假手術(shù)組,兩組差異有統(tǒng)計(jì)學(xué)( P＜0.05)；DFO組，3h輕微表達(dá)，12h明顯升高, 24h和48h仍保持一定的表達(dá)水平，7d開始降低,DFO組各時(shí)間點(diǎn)Caspase3表達(dá)低于HIBD組，兩組差異有統(tǒng)計(jì)學(xué)意義（P＜0．05）。TUNEL陽(yáng)性細(xì)胞(AI指數(shù))表達(dá)主要分布于皮層及海馬區(qū)。假手術(shù)組TUNEL陽(yáng)性細(xì)胞極少，HIBD組TUNEL陽(yáng)性細(xì)胞較對(duì)照組增多,且隨著時(shí)間延長(zhǎng)逐漸增多；兩組比較差異有統(tǒng)計(jì)學(xué)意義（P0.05）。DFO組TUNEL陽(yáng)性細(xì)胞比HIBD組明顯降低，兩組比較差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論：（1）新生大鼠給予去鐵胺后HIBD的可減輕腦組織的病理改變，提示去鐵胺對(duì)新生大鼠HIBD可能具有神經(jīng)保護(hù)作用。（2）去鐵胺可能通過(guò)上調(diào)HIF-1α的表達(dá)，降低HIBD后Caspase-3的表達(dá)，減輕HIBD病理?yè)p傷過(guò)程，抑制神經(jīng)細(xì)胞凋亡，從而對(duì)新生大鼠缺氧缺血引起的神經(jīng)細(xì)胞損傷起到一定的保護(hù)作用，進(jìn)一步為臨床治療新生兒缺氧缺血性腦損傷提供新的治療思路。
[Abstract]:Objective : To observe the protective mechanism of hypoxia - inducible factor 1偽 ( hypoxia - inducible factor 1偽 ) and deferrioxamine ( DFO ) in neonatal hypoxic - ischemic brain damage ( HIBD ) and to provide a theoretical basis for the clinical application of hypoxic - ischemic encephalopathy ( HIE ) .
Methods : 120 Wistar rats were randomly divided into sham operation group ( 8 rats ) , HIBD model group and deferrioxamine ( DFO ) group . The rats were divided into 7 subgroups : 3h group , 6h group , 12h group , 24h group , 48h group , 3d group and 7d group .
Results : ( 1 ) Different degrees of behavioral abnormalities were observed in neonatal rats after hypoxia ischemia . ( 2 ) There were some abnormal changes in brain tissue at various time points after hypoxia ischemia .
The expression of HIF - 1偽 and Caspase - 3 was significantly lower than HIBD group . The positive staining of HIF - 1偽 and Caspase - 3 was found in the cerebral cortex and hippocampus .
HIF - 1偽 positive expression was found at all time points in HIBD model group , and the expression of HIF - 1偽 was significantly increased in the same time point compared with sham operation group ( P0.05 ) . HIF - 1偽 increased at 3h after HIBD , peaked at 12h , and then decreased gradually ;
Compared with HIBD group , the expression level of HIF - 1偽 in DFO group was significantly higher than that in HIBD group ( P0.05 ) .
Caspase - 3 positive expression in HIBD model group was significantly higher than that in sham operation group at 6 h , 12 h and 24 h , and the expression level was higher than that in sham operation group ( P < 0.05 ) .
The expression levels of Caspase - 3 in DFO group and DFO group were significantly higher than those in HIBD group ( P < 0.05 ) . TUNEL positive cells ( AI index ) were mainly distributed in cortex and hippocampus . TUNEL positive cells in sham operation group were very few , TUNEL positive cells in HIBD group were higher than those of control group , and gradually increased with time .
Compared with HIBD group , TUNEL positive cells in DFO group were significantly lower than those in HIBD group ( P0.05 ) .
Conclusion : ( 1 ) In neonatal rats , HIBD can reduce the pathological changes of HIBD . It is suggested that deferrioxamine may have neuroprotection effect on neonatal rat HIBD . ( 2 ) Deferrioxamine may inhibit the expression of HIF - 1偽 , decrease the expression of Caspase - 3 after HIBD , reduce the apoptosis of HIBD , and provide a new treatment for neonatal hypoxic - ischemic brain injury .

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R722.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 湯亞南;Caspase-3與圍產(chǎn)期缺氧缺血性腦損傷[J];中國(guó)當(dāng)代兒科雜志;2002年01期

2 崔秀玉，呂國(guó)蔚;腦缺氧和腦缺氧適應(yīng)時(shí)能量代謝的變化[J];首都醫(yī)科大學(xué)學(xué)報(bào);1996年02期

3 李麗華;孔祥勇;李培國(guó);馬鳳云;郝冰;孔令凱;封志純;;鐵離子螯合劑對(duì)缺氧缺血性腦損傷新生大鼠腦組織缺氧誘導(dǎo)因子1_α表達(dá)的調(diào)節(jié)及機(jī)制[J];實(shí)用兒科臨床雜志;2011年18期

4 張平,張杏紅;缺血性腦損傷細(xì)胞凋亡的分子機(jī)制[J];河北職工醫(yī)學(xué)院學(xué)報(bào);2003年03期

5 黃越芳;莊思齊;梁英杰;;去鐵胺對(duì)新生大鼠缺氧缺血性腦的保護(hù)作用[J];中國(guó)病理生理雜志;2007年08期

本文編號(hào)：1704267