本文關(guān)鍵詞：珠蛋白在缺血及氧化損傷條件下神經(jīng)保護(hù)作用的研究　出處：《西北農(nóng)林科技大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 腦中風(fēng) 腦缺血再灌注 血液稀釋 腦紅蛋白 珠蛋白

【摘要】：腦血管疾病是威脅我國(guó)人們健康的重大疾病之一,腦血管疾病的預(yù)防和治療已經(jīng)成為醫(yī)學(xué)界的重大難題。缺血性腦中風(fēng)是常見(jiàn)的腦血管病之一,其病因是腦血管狹窄或栓塞,導(dǎo)致腦部血流阻斷,使腦組織缺血從而引起病理反應(yīng)。我們采用常用的大腦中動(dòng)脈阻塞模型(MCAO),研究腦缺血的治療,旨在探索治療腦缺血的新方案。同時(shí),腦缺血中氧自由基是造成損傷的重要來(lái)源,我們利用多重PCR技術(shù)檢測(cè)在自由基氧化環(huán)境下線蟲中33個(gè)珠蛋白(Globin)的表達(dá)差異,旨在尋找清除自由基的新藥品,為腦缺血提供新的治療方案。 血液稀釋通過(guò)降低紅細(xì)胞壓積和血液粘稠度以改善微循環(huán)和組織供氧,能抑制缺血再灌注引起的腦損傷。為研究擴(kuò)容性血液稀釋對(duì)腦缺血再灌注損傷的影響。我們以成年雄性SD大鼠為對(duì)象,制備腦缺血再灌注損傷模型。以生理鹽水為血液稀釋劑,對(duì)模型大鼠進(jìn)行擴(kuò)容性血液稀釋。在再灌注同時(shí),通過(guò)尾靜脈分別注射0.5、1.0、2.0ml的生理鹽水進(jìn)行擴(kuò)容性血液稀釋,用2, 3, 5-三苯基四氮唑(TTC)染色,然后以圖像分析方法計(jì)算TTC染色面積和腦梗死體積。結(jié)果顯示隨著注射生理鹽水體積的增加,腦梗死體積呈降低的趨勢(shì),注射2.0ml生理鹽水組腦梗死體積顯著低于對(duì)照組。說(shuō)明擴(kuò)容性血液稀釋能顯著減小大鼠腦缺血再灌注模型腦梗死體積,對(duì)大鼠腦缺血再灌注損傷具有保護(hù)作用。 腦紅蛋白(NGB)是新發(fā)現(xiàn)的在脊椎動(dòng)物神經(jīng)系統(tǒng)中特異表達(dá)的攜氧珠蛋白家族的成員,研究發(fā)現(xiàn)腦缺血能促進(jìn)腦紅蛋白的表達(dá),同時(shí)腦紅蛋白的過(guò)量表達(dá)能抑制缺血引起的腦損傷。為研究腦紅蛋白對(duì)腦缺血再灌注損傷的影響。我們以成年雄性SD大鼠為對(duì)象,制備腦缺血再灌注損傷模型。用NGB和TAT-NGB(融合導(dǎo)肽的NGB)對(duì)腦缺血再灌注模型大鼠進(jìn)行干預(yù)。于再灌注同時(shí),通過(guò)尾靜脈分別注射不同濃度的NGB和TAT-NGB溶液。用2, 3, 5-三苯基四氮唑(TTC)染色,然后以圖像分析方法計(jì)算TTC染色面積和腦梗死體積。結(jié)果顯示注射NGB和TAT-NGB溶液的大鼠腦梗死體積相對(duì)對(duì)照組減小,在NGB劑量為24mg/kg和TAT-NGB劑量為10mg/kg時(shí)腦梗死體積顯著減小。說(shuō)明NGB能減小大鼠腦缺血再灌注模型的腦梗死體積,對(duì)缺血再灌注引起的大鼠腦損傷有保護(hù)作用。 珠蛋白是幾乎存在于各種生物中的氧結(jié)合蛋白,其共同的特點(diǎn)是都含有典型的螺旋三明治折疊結(jié)構(gòu)。線蟲體內(nèi)存在33種珠蛋白,這些蛋白的功能還不清楚。為了研究線蟲體內(nèi)33種珠蛋白與活性氧自由基的關(guān)系。我們以N2野生型線蟲為實(shí)驗(yàn)對(duì)象,飼養(yǎng)于含有不同濃度(0.3 mmol/L、0.4 mmol/L、0.5 mmol/L )百草枯(PQ)的線蟲培養(yǎng)基(NGM)上,五天后,提取線蟲的總RNA,再進(jìn)行反轉(zhuǎn)錄。然后利用多重PCR技術(shù)擴(kuò)增線蟲33種珠蛋白,用凝膠定量分析軟件計(jì)算各條帶的積分光密度,計(jì)算出各個(gè)珠蛋白的相對(duì)表達(dá)量。通過(guò)與對(duì)照組比較,得出33種珠蛋白的表達(dá)變化。結(jié)果顯示在PQ處理下表達(dá)增加的基因有:Globin-7、Globin -12、Globin -19、Globin -22、Globin -24、Globin -31。這些基因在PQ的壓力下表達(dá)明顯增高,這些蛋白具有抑制PQ引起的氧化損傷的潛在能力。
[Abstract]:Cerebrovascular disease is one of the major diseases threatening people's health in our country, the prevention and treatment of cerebrovascular disease has become a major problem in the medical field. The ischemic stroke is one of the most common cerebrovascular disease, its etiology is cerebral vascular stenosis or embolism, resulting in blocking blood flow to the brain, the cerebral ischemia causing pathological reaction. We use the brain artery occlusion model (MCAO), the treatment of cerebral ischemia, to explore the new scheme for the treatment of cerebral ischemia. At the same time, the oxygen free radicals in cerebral ischemia is an important source of damage, we use multiple PCR technique in detection of free radical oxidation environment line 33 globins in C.elegans (Globin) expression the difference, in order to find a new drug to eliminate free radicals, provide a new treatment for cerebral ischemia.
Hemodilution by lowering hematocrit and blood viscosity to improve microcirculation and tissue oxygen, can inhibit brain injury induced by ischemia-reperfusion. To study the effects of hypervolemic hemodilution on cerebral ischemia reperfusion injury. We object to adult male SD rats, the preparation of the model of cerebral ischemic reperfusion injury. Saline for blood thinners, expansion of blood dilution on the rat model of reperfusion. In the meanwhile, normal saline were injected through the tail vein of 0.5,1.0,2.0ml of hypervolemic hemodilution. By 2, 3, three (TTC) 5- phenyl tetrazole staining and image analysis method to calculate TTC staining area and infarct volume. Show that with the increase of saline injection volume, cerebral infarction volume was decreased, 2.0ml saline was injected into the cerebral infarction volume was significantly lower than the control group. The expansion of the blood dilution significantly The reduction of cerebral ischemia reperfusion model cerebral infarction volume in rats has a protective effect on cerebral ischemia reperfusion injury in rats.
Neuroglobin (NGB) is a new found in the vertebrate nervous system specific expression of the globin family members carrying oxygen, the study found that cerebral ischemia can promote the expression of neuroglobin, brain damage and excessive expression of neuroglobin can inhibit ischemia induced. In order to study the effect of neuroglobin on cerebral ischemia reperfusion injury. We object to adult male SD rats, preparation of cerebral ischemia reperfusion injury model. Using NGB and TAT-NGB (fusion leader peptide NGB) on cerebral ischemia reperfusion rat model of reperfusion intervention. At the same time, through the tail vein were injected different concentrations of NGB and TAT-NGB solution. In 2, 3, three (TTC) 5- phenyl tetrazole staining and image analysis method to calculate TTC staining area and infarct volume. The results showed that the volume of cerebral infarction in rats injected with NGB and TAT-NGB solution relative to the control group decreased at the dose of NGB 24mg/ and TAT-NGB kg When the dose was 10mg/kg, the volume of cerebral infarction decreased significantly, indicating that NGB could reduce the volume of cerebral infarction in rats with cerebral ischemia-reperfusion and protect the brain injury induced by ischemia-reperfusion.
The pearl protein is present in almost all kinds of organisms oxygen binding protein, its common feature is to contain helical sandwich structure. There are 33 kinds of typical globins in C.elegans, the function of these proteins is not clear. In order to study the relationship between 33 kinds of nematode globin and active oxygen free radicals. We used N2 wild nematodes as the experimental object, raised in containing different concentrations (0.3 mmol/L, 0.4 mmol/L, 0.5 mmol/L) of paraquat (PQ) medium nematode (NGM), five days later, the total RNA extraction of nematodes, reverse transcription was then amplified by multiplex PCR. 33 species of nematode globin, calculate the integral light the density of each band by gel quantitative analysis software, calculated the relative expression of each globin. Compared with the control group, the expression obtained 33 globin. The results showed that the increase of gene expression under PQ treatment: Globin-7, Globin -12, Globin -19, Globin -22, Globin -24, Globin -31. have significantly higher expression levels under PQ pressure. These proteins have potential to inhibit oxidative damage induced by PQ.

【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 陳婷方;侯冰;任長(zhǎng)虹;高艷;吳永紅;吳祖澤;張成崗;;腦紅蛋白減輕硝普鈉誘導(dǎo)的PC12細(xì)胞損傷[J];軍事醫(yī)學(xué)科學(xué)院院刊;2008年04期

2 任長(zhǎng)虹;張成崗;;秀麗隱桿線蟲在神經(jīng)科學(xué)研究中的應(yīng)用[J];軍事醫(yī)學(xué)科學(xué)院院刊;2008年06期

3 張榮媛;張宏偉;尚愛(ài)加;;大鼠大腦中動(dòng)脈缺血再灌注模型制備及行為學(xué)評(píng)價(jià)標(biāo)準(zhǔn)的改進(jìn)[J];軍醫(yī)進(jìn)修學(xué)院學(xué)報(bào);2008年05期

4 吳永紅;張成崗;;HIV-1TAT蛋白轉(zhuǎn)導(dǎo)肽的研究進(jìn)展[J];中國(guó)生物工程雜志;2010年10期

5 褚曉凡,董家政,吳軍,饒明俐,張淑琴;頸內(nèi)動(dòng)脈線栓與環(huán)扎建立大鼠局灶腦缺血再灌模型[J];中風(fēng)與神經(jīng)疾病雜志;2000年03期

6 劉亢丁,蘇志強(qiáng),李毅平,饒明俐;實(shí)驗(yàn)性局灶性腦缺血再灌注動(dòng)物模型的改進(jìn)及評(píng)價(jià)[J];中風(fēng)與神經(jīng)疾病雜志;1997年02期

7 吳偉,史繼新;腦缺血再灌注損傷后神經(jīng)元損傷機(jī)制及治療研究進(jìn)展[J];中國(guó)微循環(huán);2003年06期

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