本文選題：顱腦外傷 + 神經(jīng)生長(zhǎng)因子��； 參考：《南京大學(xué)》2013年碩士論文

【摘要】：顱腦外傷(traumatic brain injury, TBI)是青年人出現(xiàn)神經(jīng)功能障礙最常見(jiàn)的原因之一,TBI引起的繼發(fā)性損傷較原發(fā)性損傷更為嚴(yán)重,包括腦水腫、阿爾茨海默病(Alzheimer's disease, AD)等,造成患者預(yù)后不良。神經(jīng)生長(zhǎng)因子(nerve growth factor, NGF)具有修復(fù)受損組織的潛力,可以改善TBI大鼠的預(yù)后,但由于血腦屏障的作用,其臨床使用方面受到了限制,而經(jīng)鼻給藥的方式有效的解決了這一難題。本研究分別從腦水腫和AD兩方面入手,旨在探討經(jīng)鼻給予NGF改善TBI大鼠預(yù)后的潛在機(jī)制。研究將成年雄性Sprague-Dawley大鼠分為T(mén)BI組、NGF+TBI組和control組,參照Feeney自由落體損傷的方法制作TBI模型,NGF+TBI組大鼠經(jīng)鼻給予NGF(50ul/d),TBI組大鼠經(jīng)鼻給予等量的磷酸鹽緩沖液。采用干濕重評(píng)估腦組織含水量,免疫組織化學(xué)法檢測(cè)腦組織內(nèi)水通道蛋白-4(aquaporin-4,AQP4)、tau蛋白的表達(dá)情況,取患側(cè)腦皮質(zhì),western blotting法測(cè)定AQP4、tau蛋白、糖原合成酶激酶-3β(glycogen synthase kinase-3β, GSK-3β)的水平,ELISA法、qRT-PCI法測(cè)定白介素-1β (interleukin-1β, IL-1β)的蛋白及基因的表達(dá)水平。EMSA法測(cè)定腦組織內(nèi)核因子-κB (nuclear factor-κB, NF-κB)的DNA結(jié)合活力。qRT-PCR法測(cè)定B淋巴細(xì)胞瘤-2(B-celllymphoma-2, Bcl-2)、蛋白水解酶-3(cysteinyl aspartate specific proteinase-3, caspase-3)的基因表達(dá)情況。與control組相比,TBI組大鼠腦組織含水量明顯增高,腦皮質(zhì)內(nèi)AQP4的表達(dá)明顯增高,經(jīng)NGF治療后,大鼠的腦組織含水量及AQP4的表達(dá)明顯下降；TBI后,大鼠腦組織內(nèi)tau蛋白、GSK-3β的表達(dá)較control組大鼠明顯增加,UNGF+TBI組大鼠tau蛋白、GSK-3β的表達(dá)較TBI組明顯下降。進(jìn)一步研究結(jié)果提示,TBI后大鼠IL-1β的蛋白及基因水平均明顯升高,NF-κB的DNA結(jié)合活性明顯增加,Bcl-2的基因水平降低,caspase-3的基因水平升高,經(jīng)鼻給予NGF后IL-1β的蛋白及基因水平均明顯下降,NF-κB的DNA結(jié)合活性明顯降低,Bcl-2的基因水平升高,caspase-3的基因水平降低。由此可推,經(jīng)鼻給予NGF可以減輕AQP4相關(guān)的腦水腫,減少GSK-3β關(guān)聯(lián)的tau蛋白的磷酸化,從而改善TBI大鼠的預(yù)后,以上兩個(gè)作用均與NGF減輕了TBI后Bcl-2/caspase-3/NF-κB通路相關(guān)的炎癥反應(yīng)有關(guān)。
[Abstract]:Traumatic brain injury, TBI) is one of the most common causes of neurologic dysfunction in young people. The secondary injury caused by TBI is more serious than that caused by primary injury, including brain edema, Alzheimer's disease and so on, resulting in poor prognosis of the patients. Nerve growth factor (NGF) nerve growth factor, NGF) has the potential to repair damaged tissue and can improve the prognosis of TBI rats. However, the clinical use of nerve growth factor growth factor, NGF) is limited due to the effect of blood-brain barrier, and the method of nasal administration can effectively solve this problem. The purpose of this study was to explore the potential mechanism of nasal administration of NGF in improving the prognosis of TBI rats from the aspects of brain edema and AD. Adult male Sprague-Dawley rats were divided into TBI group and control group. According to the method of Feeney free falling body injury, the TBI model rats in NGF TBI group were given the same amount of phosphate buffer by nasal administration. The water content of brain tissue was evaluated by wet and dry weight, the expression of aquaporin-4 aquaporin-4aqp4ttau protein in brain tissue was detected by immunohistochemical method, and the tau protein was detected by western blotting method in the affected cerebral cortex. Expression of interleukin-1 尾 -interleukin-1 尾 (IL-1 尾) protein and gene by qRT-PCI. DNA binding activity of NF- 魏 B nuclear factor- 魏 B, NF- 魏 B, NF- 魏 B in brain tissue was assayed by DNA binding activity. QRT-PCR was used to detect B-cell lymphoma-2, Bcl-2U. Gene expression of protein hydrolase-3 aspartate specific proteinase-3 (caspase-3). Compared with the control group, the water content of brain tissue and the expression of AQP4 in the cortex of the rats were significantly increased. After NGF treatment, the water content and the expression of AQP4 in the brain tissue of the rats decreased significantly. The expression of tau protein GSK-3 尾 in rat brain was significantly higher than that in control group. The expression of tau protein GSK-3 尾 in UNGF TBI group was significantly lower than that in TBI group. The results showed that the protein and gene level of IL-1 尾 increased significantly after TBI in rats. The DNA binding activity of NF- 魏 B increased significantly, and the gene level of Bcl 2 decreased the level of caspase-3. The protein and gene level of IL-1 尾 decreased significantly after nasal administration of NGF. The DNA binding activity of NF- 魏 B decreased significantly. The level of Bcl-2 gene increased and the gene level of caspase-3 decreased. Therefore, nasal administration of NGF can reduce the brain edema associated with AQP4, decrease the phosphorylation of tau protein associated with GSK-3 尾, and improve the prognosis of TBI rats. Both of these effects are related to the reduction of inflammation related to Bcl-2 / caspase-3 / NF- 魏 B pathway after TBI by NGF.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R651.15

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

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本文編號(hào)：1826181