【摘要】：背景:多發(fā)性硬化(multiple sclerosis,MS)是一種中樞神經(jīng)系統(tǒng)(central nerve system,CNS)的慢性自身免疫性疾病,以炎癥復(fù)發(fā)和病情緩解交替發(fā)生為特征,好發(fā)于15~50歲的青壯年之中。目前臨床中對MS患者大都采用綜合性治療的方式,但是即便治療及時,這些療法也只能起到緩解而非治愈MS的效果。因此,增強(qiáng)MS的治療康復(fù),已經(jīng)成為臨床及基礎(chǔ)研究中的熱點與重點。鋰劑是臨床治療預(yù)防雙相情感障礙躁狂的一線藥物,具備對中樞神經(jīng)系統(tǒng)的保護(hù)功能,同時能夠?qū)Χ喾N免疫細(xì)胞產(chǎn)生調(diào)節(jié)作用。目前的研究表明,使用鋰劑能夠在實驗動物中對MS替代模型實驗性自身免疫性腦脊髓炎(experimental autoimmune encephalomyelitis,EAE)產(chǎn)生顯著的治療作用,并且該治療作用和IFN-γ干擾素γ(interferon-γ,IFN-γ)存在一定的聯(lián)系。IFN-γ由Th1型細(xì)胞和單核細(xì)胞合成分泌,可以調(diào)節(jié)細(xì)胞因子之間的“級聯(lián)作用模式”,增強(qiáng)免疫應(yīng)答,在MS病人血清中表達(dá)升高。除了關(guān)注鋰劑對于免疫系統(tǒng)的作用以外,我們還推測,鋰劑治療MS的過程,是一個抑制GSK3β、進(jìn)而激活神經(jīng)細(xì)胞內(nèi)促細(xì)胞生存Wnt/β-catenin通路的過程。綜上,探究鋰劑對MS具體的治療效果及相關(guān)機(jī)制,有助于提升MS病人的預(yù)后,同時發(fā)掘更多的治療靶點。在本項研究中,我們重點關(guān)注鋰劑在IFN-γ功能缺失時對CNS炎癥的影響,同時還探究了β-catenin在鋰劑治療MS中扮演的具體角色。目的:探究鋰劑在治療MS過程中與IFN-γ和β-catenin的相互作用。方法:1.IFN-γR缺失小鼠PCR鑒定;2.使用MOG35-55和百日咳毒素聯(lián)合注射的方法,誘導(dǎo)IFN-γR缺失小鼠發(fā)生EAE;3.對不同組中EAE小鼠腦部炎癥非典型癥狀比例進(jìn)行統(tǒng)計;4.對不同組中EAE小鼠脊髓炎癥典型癥狀進(jìn)行打分統(tǒng)計;5.收集小鼠腦和脊髓組織,進(jìn)行HE染色分析;6.氯化鋰和IFN-γ處理小鼠腦內(nèi)皮細(xì)胞b END.3;7.免疫印跡檢測b END.3細(xì)胞中β-catenin表達(dá)量的變化;結(jié)果:1.經(jīng)PCR驗證,實驗用小鼠為IFN-γR-/-基因型,為IFN-γR缺失;2.MOG35-55和百日咳毒素聯(lián)合注射的方法,成功在IFN-γR缺失小鼠中誘導(dǎo)了腦和脊髓炎癥的發(fā)生;3.鋰劑治療降低了腦部炎癥非典型癥狀的比例以及脊髓炎癥典型癥狀的平均得分;4.鋰劑治療緩解了腦和脊髓組織中的炎癥反應(yīng);5.鋰劑對于CNS中炎癥的治療作用,可以不依賴于IFN-γ的功能;6.鋰劑對于IFN-γR缺失小鼠腦和脊髓炎癥的緩解作用,不具有長效性;7.鋰劑和IFN-γ處理,顯著提高了腦內(nèi)皮細(xì)胞中β-catenin的表達(dá)水平;結(jié)論:鋰劑可以激活腦內(nèi)皮細(xì)胞中β-catenin信號,抑制腦和脊髓炎癥的發(fā)生;鋰劑對于CNS炎癥的治療,可以不依賴于IFN-γ的功能,但相應(yīng)治療作用不具有長效性。
[Abstract]:Background: multiple sclerosis (multiple sclerosis,MS) is a chronic autoimmune disease of central nervous system (central nerve system,CNS) characterized by recurrent inflammation and remission. At present, MS patients are mostly treated by comprehensive therapy, but even if the treatment is timely, these therapies can only alleviate rather than cure MS. Therefore, enhancing the treatment and rehabilitation of MS has become the focus of clinical and basic research. Lithium is a first-line drug for the prevention of bipolar affective disorder mania. It can protect the central nervous system and regulate many immune cells. Current studies have shown that the use of lithium can produce significant therapeutic effects on experimental autoimmune encephalomyelitis (experimental autoimmune encephalomyelitis,EAE) induced by MS in experimental animals, and the therapeutic effect is similar to that of IFN- interferon 緯 (interferon- 緯). IFN- 緯 was synthesized and secreted by Th1 type cells and monocytes, which could regulate the "cascade action mode" between cytokines, enhance the immune response, and increase the expression of IFN- 緯 in serum of MS patients. In addition to focusing on the role of lithium in the immune system, we also speculate that the treatment of MS with lithium is a process that inhibits GSK3 尾 and thus activates the Wnt/ 尾-catenin pathway that promotes cell survival in nerve cells. In conclusion, to explore the specific therapeutic effects and related mechanisms of lithium on MS is helpful to improve the prognosis of patients with MS, and to explore more therapeutic targets at the same time. In this study, we focused on the effects of lithium on CNS inflammation in the absence of IFN- 緯 function, and explored the role of 尾-catenin in the treatment of MS with lithium. Aim: to investigate the interaction of lithium with IFN- 緯 and 尾-catenin in the treatment of MS. Methods: 1. PCR identification of IFN- 緯 R deficient mice. Induction of EAE;3. in IFN- 緯 R deficient mice by combined injection of MOG35-55 and pertussis toxin The proportion of atypical symptoms of cerebral inflammation in EAE mice was analyzed in different groups. 4. The typical symptoms of spinal cord inflammation in EAE mice in different groups were evaluated. The brain and spinal cord of mice were collected and analyzed by HE staining. Treatment of b END.3;7. in mouse brain endothelial cells with lithium chloride and IFN- 緯 The expression of 尾-catenin in b END.3 cells was detected by Western blotting. Results: 1. PCR showed that the mice were IFN- 緯 R-r- genotype and IFN- 緯 R deletion, and the combination of 2.MOG35-55 and pertussis toxin induced the inflammation of brain and spinal cord in IFN- 緯 R deficient mice. 3. Lithium treatment decreased the proportion of atypical symptoms of cerebral inflammation and the average score of typical symptoms of spinal cord inflammation; 4. 5%. Lithium treatment alleviated inflammation in brain and spinal cord. The therapeutic effect of lithium on inflammation in CNS could not be dependent on the function of IFN- 緯. The effect of lithium on brain and spinal cord inflammation in mice with IFN- 緯 R deficiency was not long-lasting. Lithium and IFN- 緯 could significantly increase the expression of 尾-catenin in brain endothelial cells. Conclusion: lithium can activate 尾-catenin signal in brain endothelial cells and inhibit the occurrence of inflammation in brain and spinal cord. Lithium may not depend on the function of IFN- 緯 in the treatment of CNS inflammation, but the corresponding therapeutic effect is not long-lasting.
【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R744.51

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