【摘要】：以往關(guān)于脊髓損傷的研究,不管是以實(shí)驗(yàn)鼠或斑馬魚等為模式動(dòng)物開展的基礎(chǔ)性研究,還是臨床上對(duì)于治療脊髓損傷的新方法、新藥物的嘗試,基本都只關(guān)注受損的脊髓本身。本課題中,我們不再局限于受損的脊髓組織,而是從腦內(nèi)的單胺類神經(jīng)遞質(zhì)組胺(histamine,HA)著手,將同是中樞神經(jīng)系統(tǒng)的腦組織同脊髓損傷聯(lián)系起來(lái),以期探究腦源性信號(hào)對(duì)下游脊髓組織修復(fù)的影響,發(fā)現(xiàn)影響脊髓損傷修復(fù)的新機(jī)制。作為一種來(lái)源于腦組織并下行至脊髓的單胺能神經(jīng)遞質(zhì),組胺在脊髓損傷修復(fù)中作用的報(bào)道較少;而已有研究表明腦內(nèi)單胺能神經(jīng)遞質(zhì)信號(hào)的多巴胺(dopamine,DA)和5-羥色胺(5-hydroxytryptamine,5-HT)具有重要的運(yùn)動(dòng)調(diào)節(jié)功能,對(duì)斑馬魚脊髓損傷后的修復(fù)有積極作用。我們采用成年斑馬魚,首先向其間腦內(nèi)注射組胺,其后進(jìn)行脊髓損傷手術(shù),建立腦源性組胺水平升高的脊髓損傷斑馬魚模型,向?qū)φ战M斑馬魚注射生理鹽水并進(jìn)行脊髓損傷。行為學(xué)分析的結(jié)果顯示,間腦內(nèi)組胺注射可以抑制脊髓損傷斑馬魚運(yùn)動(dòng)功能的恢復(fù)。免疫熒光染色結(jié)果顯示,在脊髓損傷后的不同時(shí)間點(diǎn),間腦組胺注射對(duì)損傷位點(diǎn)神經(jīng)膠質(zhì)細(xì)胞產(chǎn)生影響:損傷后第3天,脊髓損傷位點(diǎn)上段的小膠質(zhì)細(xì)胞的熒光強(qiáng)度增強(qiáng),第7天恢復(fù)至對(duì)照組水平;而對(duì)于放射狀膠質(zhì)細(xì)胞,是在損傷后第7天促進(jìn)其增殖,在第11天恢復(fù)至對(duì)照組水平,但是,在損傷后第11天,損傷位點(diǎn)上段放射狀膠質(zhì)細(xì)胞呈現(xiàn)出不利于損傷后修復(fù)的多極星狀形態(tài)。此外,組胺注射還加劇了下行到脊髓損傷位點(diǎn)的酪氨酸羥化酶陽(yáng)性和5-羥色胺陽(yáng)性神經(jīng)纖維的崩解。實(shí)時(shí)定量聚合酶鏈?zhǔn)椒磻?yīng)(qRT-PCR)的結(jié)果表明,斑馬魚間腦組胺注射使損傷位點(diǎn)上段的腦源性神經(jīng)營(yíng)養(yǎng)因子(BDNF)在損傷后第1天和第3天的表達(dá)減少;使胰島素樣生長(zhǎng)因子1(IGF-1)在損傷后第3天的表達(dá)下調(diào)。以上實(shí)驗(yàn)結(jié)果表明,斑馬魚間腦組胺注射可以抑制成年斑馬魚脊髓損傷后運(yùn)動(dòng)功能的恢復(fù),其機(jī)制可能是通過(guò)調(diào)節(jié)損傷位點(diǎn)的神經(jīng)營(yíng)養(yǎng)及免疫微環(huán)境,以及促進(jìn)從腦組織下行到脊髓的其他單胺能神經(jīng)纖維的崩解。本課題以腦源性信號(hào)—組胺為研究對(duì)象,探究了其對(duì)下游脊髓組織修復(fù)的影響,為發(fā)現(xiàn)脊髓損傷修復(fù)的新機(jī)制提供了獨(dú)特思路。
[Abstract]:Previous studies on spinal cord injury, whether based on experimental rats or zebrafish as model animals, or clinical treatment of new methods of spinal cord injury, new drugs, basically only focus on the injured spinal cord itself. In this study, we no longer confine ourselves to the injured spinal cord, but start with the monoamine neurotransmitter histamine HA in the brain, linking the same brain tissue of the central nervous system to the spinal cord injury. The purpose of this study was to explore the effects of brain-derived signals on the repair of the downstream spinal cord, and to find out the new mechanism that affects the repair of spinal cord injury. As a monoaminergic neurotransmitter derived from brain tissue and descending to the spinal cord, histamine plays an important role in the repair of spinal cord injury. Some studies have shown that dopamine DA and 5-hydroxytryptamine 5-HT in the brain of monoaminergic neurotransmitter signal play an important role in motor regulation and play a positive role in the repair of spinal cord injury in zebrafish. In adult zebrafish, histamine was injected into the brain at first, then spinal cord injury was performed. The model of spinal cord injury was established with elevated histamine level. The normal saline was injected into the control group and spinal cord injury was carried out. Behavioral analysis showed that diencephalon histamine injection inhibited the recovery of motor function in spinal cord injury zebrafish. The results of immunofluorescence staining showed that at different time points after spinal cord injury, diencephalon histamine injection had an effect on the glial cells at the injured site: the fluorescence intensity of the microglia in the upper segment of the spinal cord injury site increased on the 3rd day after injury. On the 7th day, the radiative glial cells increased their proliferation on the 7th day after injury and recovered to the control level on the 11th day, but on the 11th day after the injury, the radiative glial cells returned to the level of the control group, but on the 11th day after the injury, The radial glial cells in the upper segment of injury site showed multipolar stellate morphology which was not conducive to repair after injury. In addition, histamine injection increased the disintegration of tyrosine hydroxylase positive and serotonin positive nerve fibers descending to the site of spinal cord injury. The results of real-time quantitative polymerase chain reaction (qRT-PCR) showed that diencephalon histamine injection decreased the expression of brain-derived neurotrophic factor (BDNF) in the upper part of the injury site on the 1st and 3rd day after injury. The expression of insulin-like growth factor 1 (IGF-1) was down-regulated on the 3rd day after injury. These results suggest that diencephalon histamine injection can inhibit the recovery of motor function after spinal cord injury in adult zebrafish, and its mechanism may be by regulating the neurotrophic and immune microenvironment of the injured site. And promote the disintegration of other monoaminergic nerve fibers descending from brain tissue to spinal cord. In this study, the effects of brain-derived signal histamine on the repair of the downstream spinal cord were investigated, which provided a unique idea for discovering the new mechanism of spinal cord injury repair.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R651.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Melitta Schachner;;A Surgery Protocol for Adult Zebrafish Spinal Cord Injury[J];遺傳學(xué)報(bào);2012年09期

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本文編號(hào)：2176433