本文選題：膠質(zhì)瘢痕 + 星形膠質(zhì)細(xì)胞　； 參考：《吉林大學(xué)》2015年博士論文

【摘要】：背景： 反應(yīng)性膠質(zhì)增生，亦稱為膠質(zhì)瘢痕形成，是中樞神經(jīng)系統(tǒng)在受到損傷后繼發(fā)的一種特殊的細(xì)胞反應(yīng)。此病理過(guò)程的主要參與者包括激活的星形膠質(zhì)細(xì)胞，激活的小膠質(zhì)細(xì)胞和細(xì)胞外間質(zhì)成分。在中樞神經(jīng)損傷發(fā)生后，損傷急性期炎癥反應(yīng)所生成的化學(xué)因子和細(xì)胞因子通過(guò)血-腦屏障或血-脊髓屏障進(jìn)入神經(jīng)系統(tǒng)，先后將小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞激活，最終導(dǎo)致膠質(zhì)增生的發(fā)生。在此過(guò)程中生成的膠質(zhì)瘢痕一方面可以局限炎癥反應(yīng)的范圍，支持損傷組織，減輕細(xì)胞毒性反應(yīng)；另一方面它卻可以阻礙神經(jīng)再生和神經(jīng)髓鞘再生。但是，激活反應(yīng)先行膠質(zhì)增生的機(jī)制目前尚不明確。血小板激活因子（PAF）是一種內(nèi)源性磷脂多能性介質(zhì)，它在免疫系統(tǒng)和神經(jīng)系統(tǒng)中都發(fā)揮著重要作用。一方面它可以維持細(xì)胞的正常生理活動(dòng)，另一方面它可以造成病理反應(yīng)。研究已經(jīng)證明過(guò)量表達(dá)的PAF與許多中樞神經(jīng)系統(tǒng)的疾病都有關(guān)系，而應(yīng)用PAF拮抗劑可以緩解一部分神經(jīng)系統(tǒng)疾病的癥狀。在本研究中我們假設(shè)PAF信號(hào)通路可以在脊髓損傷后啟動(dòng)反應(yīng)性膠質(zhì)增生，而阻斷PAF信號(hào)通路則可以抑制膠質(zhì)瘢痕形成，改善損傷后神經(jīng)功能恢復(fù)情況。 研究目的： 通過(guò)實(shí)驗(yàn)探究阻斷PAF信號(hào)通路對(duì)小鼠脊髓損傷后反應(yīng)性膠質(zhì)增生和神經(jīng)功能恢復(fù)的影響。 實(shí)驗(yàn)方法： 測(cè)量體重、行為學(xué)等指標(biāo)證明WT和PAFR敲除小鼠在大體發(fā)育上沒(méi)有顯著差異；用免疫熒光染色和Western Blot觀察WT和PAFR敲除小鼠的脊髓族中內(nèi)星形膠質(zhì)細(xì)胞和小膠質(zhì)細(xì)胞的含量與分布；在WT小鼠脊髓中注射外源性PAF觀察PAF對(duì)星形膠質(zhì)細(xì)胞及小膠質(zhì)細(xì)胞增殖、激活的影響；用Rotarod test和Griptest等行為學(xué)實(shí)驗(yàn)測(cè)量脊髓損傷的PAFR敲除小鼠在損傷后特定時(shí)間點(diǎn)的神經(jīng)功能恢復(fù)情況；用Western Blot和免疫熒光染色測(cè)量和觀察脊髓損傷后PAFR敲除小鼠脊髓組織內(nèi)各種細(xì)胞因子如白細(xì)胞介素-6、特定蛋白與中間絲蛋白如GFAP的表達(dá)量和分布情況，從而說(shuō)明小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞在脊髓受損傷后的激活情況；用免疫熒光染色觀察脊髓損傷之后小鼠的神經(jīng)軸突脫髓鞘/再生的情況；用行為學(xué)實(shí)驗(yàn)測(cè)量對(duì)WT損傷小鼠系統(tǒng)性地使用PAFR拮抗劑治療后小鼠神經(jīng)功能恢復(fù)情況。 結(jié)果： （a）小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞可以被外源性PAF劑量依賴性激活；（b）在脊髓損傷后，，PAFR敲除小鼠的神經(jīng)功能恢復(fù)要顯著優(yōu)于WT小鼠；（c）雖然在脊髓損傷后PAFR敲除小鼠和WT小鼠的小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞都被激活，但是PAFR敲除小鼠脊髓組織中IL-6，GFAP，波形蛋白和硫酸軟骨蛋白多糖（CSPGs）表達(dá)量并未顯著升高；（d）PAFR小鼠在脊髓損傷后軸突回縮更少；（e）就與對(duì)照組比較而言，在亞急性期和慢性期接受PAF拮抗劑治療的WT小鼠的功能恢復(fù)程度與在急性期接受治療的WT小鼠的恢復(fù)程度相比顯著提高。 結(jié)論： PAF信號(hào)通路參與脊髓損傷后反應(yīng)性膠質(zhì)增生的激活；阻斷此信號(hào)通路可以改善損傷后的功能恢復(fù)還可以在一定程度上促進(jìn)神經(jīng)再生。
[Abstract]:Background:
Reactive gliosis, also known as glial scar formation, is a special cell response to the central nervous system secondary to injury. The main participants in this pathological process include activated astrocytes, activated microglia and extracellular matrix. After the armature injury of the middle armature, the acute inflammatory reaction is damaged. The chemical and cytokine generated by the formation of the blood brain barrier or the blood spinal cord barrier enter the nervous system and activate the microglia and astrocytes successively and eventually lead to the occurrence of glial proliferation. In this process, the formation of glial scar can limit the scope of the inflammatory reaction, support the damaged tissue, and reduce the cytotoxicity. Sexual reaction; on the other hand, it can impede nerve regeneration and regeneration of the myelin sheath. However, the mechanism that activates the activation of glial proliferation is not yet clear. Platelet activating factor (PAF) is an endogenous phospholipid pluripotent medium, which plays an important role in both the immune system and the nervous system. On the one hand, it can maintain cells. The normal physiological activity, on the other hand, can cause a pathological response. Studies have shown that excessive expression of PAF is associated with many diseases in the central nervous system, and the application of PAF antagonists can relieve some of the symptoms of a nervous system disease. In this study, we hypothesized that the PAF signal pathway could initiate reactivity after spinal cord injury. Glial hyperplasia, and blocking the PAF signaling pathway can inhibit glial scar formation and improve the recovery of nerve function after injury.
The purpose of the study is:
To explore the effects of blocking PAF signaling pathway on reactive gliosis and neurological function recovery after spinal cord injury in mice.
Experimental methods:
Measurement of weight, behavior and other indicators showed that there was no significant difference in the gross development of WT and PAFR knockout mice; the content and distribution of astrocytes and microglia in the spinal cord of WT and PAFR knockout mice were observed by immunofluorescence staining and Western Blot, and the exogenous PAF was injected into the astrocytes in the spinal cord of WT mice to observe the astrocytes. The effects of proliferation and activation of cells and microglia; measurement of neural function recovery at specific time points after injury in PAFR knockout mice with spinal cord injury by Rotarod test and Griptest; and Western Blot and immunofluorescence staining to measure and observe all kinds of cell causes in the spinal cord tissue of PAFR knockout mice after spinal cord injury. The expression and distribution of interleukin -6, specific protein and intermediate silk protein, such as GFAP, were used to demonstrate the activation of microglia and astrocytes after the injury of the spinal cord. Immunofluorescence staining was used to observe the demyelination / regeneration of the axon after spinal cord injury, and the behavior test was used to measure WT. The injured mice were systematically treated with PAFR antagonist to restore neurological function.
Result錛

本文編號(hào)：1784203