【摘要】：【目的】 建立新生4天SD大鼠腦室周圍白質(zhì)軟化模型（periventricular lerkomalacia,PVL）。在構(gòu)建PVL模型大鼠的條件下，研究HIF-1α對TfR mRNA表達的調(diào)節(jié)，為缺氧缺血腦病發(fā)生機制與腦鐵代謝之間關(guān)系的研究提供實驗基礎(chǔ)依據(jù)。 【方法】 1．4日齡新生SD大鼠通過隨機數(shù)字表隨機分為兩組：PVL組和對照組；PVL組采用分離結(jié)扎雙側(cè)頸總動脈并予缺氧的方法建立腦室周圍白質(zhì)軟化新生大鼠動物模型；對照組僅分離雙側(cè)頸總動脈，不予結(jié)扎及缺氧。采用HE染色法觀察腦組織病理學(xué)改變，Western blot法檢測MBP蛋白表達，實驗動物生長發(fā)育及神經(jīng)行為學(xué)等方法對動物模型進行評價。 2．設(shè)立（1）對照＋HIF-1α組（2）對照＋空白組（3）PVL＋抗HIF-1α組（4）PVL＋PBS組；及（1）對照＋PBS組（2）對照＋抗HIF-1α組（3）PVL＋PBS組（4）PVL＋抗HIF-1α組；以Western blot技術(shù)測HIF-1α表達，以Real-time PCR技術(shù)測TfR mRNA的表達。 【結(jié)果】 1．對4日齡新生SD大鼠進行雙側(cè)頸總動脈結(jié)扎并予以缺氧后，模型組與對照組大鼠相比，模型組大鼠生長緩慢（P0.01），睜眼時間延遲（21.67±1.19dVS15.54±1.60d），懸崖逃避能力及雙臂懸吊能力均降低，差異有統(tǒng)計學(xué)意義(P＜0.001)。在水迷宮實驗中，模型組大鼠在水中尋找站臺的游泳速度減慢、尋找站臺的所用的時間延長、撤去平臺后在平臺所在象限停留時間縮短(P＜0.05)。 2．腦組織HE染色，模型組術(shù)后24小時、48小時可見彌散性皮質(zhì)下和腦室周白質(zhì)疏松水腫。 3．Western blot法顯示，HI1周后模型組MBP表達量較對照組降低，差異具有統(tǒng)計學(xué)意義。 4．對照＋PBS組與PVL＋PBS組對比，Western blot法檢測后者HIF-1α蛋白表達增加，Real-time PCR檢測TfR mRNA的表達也增加，差異有統(tǒng)計學(xué)意義。 5．對照＋HIF-1α組與對照＋空白組對比，Western blot法檢測前者HIF-1α表達增多，Real-time PCR檢測TfR mRNA的表達也增多，差異有統(tǒng)計學(xué)意義。 6．PVL＋抗HIF-1α組與PVL＋PBS組對比，Western blot法檢測后者HIF-1α表達增多，Real-time PCR檢測TfR mRNA的表達也增多，差異有統(tǒng)計學(xué)意義。 【結(jié)論】 1．通過對4日齡新生SD大鼠進行雙側(cè)頸總動脈結(jié)扎聯(lián)合缺氧的方法，可建立與人類早產(chǎn)兒腦室周圍白質(zhì)軟化病理相似的PVL動物模型。 2．缺氧缺血PVL可見HIF-1α表達增多，TfR mRNA的表達也增多； 3．PVL模型中，抑制腦組織中HIF-1α的活性，，TfR mRNA表達量隨之減少，增加HIF-1α的表達，TfR mRNA表達量隨之增加。即可推斷HIF-1α可誘導(dǎo)TfR mRNA表達增加。 4．TfR mRNA表達的增加，可能是腦缺氧缺血后鐵代謝失衡的一個機制。由于腦鐵代謝的復(fù)雜性，HIF-1α與TfR的作用機制需進一步研究。
[Abstract]:[objective] to establish a model of periventricular leukomalacia (periventricular lerkomalacia,PVL) in neonatal SD rats. Under the condition of constructing PVL model rats, the regulation of TfR mRNA expression by HIF-1 偽 was studied, which provided an experimental basis for the study of the relationship between the pathogenesis of hypoxia-ischemic encephalopathy and brain iron metabolism. [methods] 1.4-day-old neonatal SD rats were randomly divided into two groups: PVL group and control group, and the neonatal rat model of periventricular leukomalacia was established by separating and ligating bilateral common carotid arteries and hypoxia in PVL group. In the control group, only bilateral common carotid arteries were separated without ligation and hypoxia. The pathological changes of brain tissue were observed by HE staining. The expression of MBP protein was detected by, Western blot method, and the animal model was evaluated by growth and development of experimental animals and neurobehavior. 2. Set up (1) control + HIF- 1 偽 group (2) control + blank group (3) PVL+ anti HIF- 1 偽 group (4) PVL+PBS group; And (1) control + PBS group (2) control + anti-HIF- 1 偽 group (3) PVL+PBS group (4) PVL+ anti-HIF-1 偽 group, Western blot technique was used to detect the expression of HIF-1 偽, Real-time PCR technique was used to detect the expression of TfR mRNA. [results] 1. After bilateral common carotid artery occlusion and hypoxia in 4-day-old neonatal SD rats, the growth of the model group was slower than that of the control group (P 0.01). The time of eye opening was delayed (21.67 鹵1.19dVS15.54 鹵1.60d), and the cliff escape ability and double arm suspension ability were decreased (P < 0.001). In the water maze experiment, the swimming speed of the model group rats looking for the platform in the water slowed down, the time used to find the platform was prolonged, and the residence time in the quadrant of the platform was shortened after the platform was removed (P < 0.05). 2. HE staining of brain tissue showed disseminated subcortical and periventricular white matter loose edema 24 hours and 48 hours after operation in the model group. 3.Western blot method showed that the expression of MBP in the model group was lower than that in the control group after HI1 week, and the difference was statistically significant. 4. Compared with PVL+PBS group, the expression of HIF-1 偽 protein in control + PBS group and PVL+PBS group was increased by, Western blot method, and the expression of TfR mRNA detected by Real-time PCR was also increased, the difference was statistically significant. 5. the expression of HIF-1 偽 and TfR mRNA detected by, Western blot in control + HIF-1 偽 group and control + blank group were significantly higher than those in control + HIF-1 偽 group and control + blank group, respectively. 6. Compared with PVL+PBS group, the expression of HIF-1 偽 in PVL + anti-HIF-1 偽 group was increased by, Western blot method, and the expression of TfR mRNA in PVL + anti-PVL 偽 group was also higher than that in PVL + anti-PVL 偽 group, the difference was statistically significant. [conclusion] 1. PVL animal model can be established by bilateral common carotid artery occlusion combined with hypoxia in 4-day-old neonatal SD rats, which is similar to the pathology of periventricular leukomalacia in human premature infants. 2. The expression of HIF-1 偽 in PVL increased and the expression of, TfR mRNA increased in 3.PVL model. In 3.PVL model, the expression of HIF-1 偽 decreased and the expression of, TfR mRNA increased with the increase of HIF-1 偽 expression in brain tissue. It can be inferred that HIF-1 偽 can induce the increase of TfR mRNA expression. The increase of 4.TfR mRNA expression may be a mechanism of iron metabolism imbalance after cerebral hypoxia and ischemia. Due to the complexity of brain iron metabolism, the mechanism of interaction between HIF-1 偽 and TfR needs to be further studied.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R-332

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相關(guān)期刊論文 前1條

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