本文選題：重癥肌無力 + 胸腺瘤　； 參考：《中國人民解放軍醫(yī)學(xué)院》2017年博士論文

【摘要】：正文:第一部分:伴胸腺瘤重癥肌無力患者外周血Th17細(xì)胞比例及細(xì)胞因子表達(dá)量變化目的:重癥肌無力(myasthenia gravis, MG)是T細(xì)胞輔助、乙酰膽堿受體抗體(AchR-Abs)介導(dǎo)的神經(jīng)肌肉接頭傳遞障礙的自身免疫性疾病。Th17細(xì)胞在MG發(fā)病機(jī)制中的作用仍不明確。本研究目的為探索伴胸腺瘤MG患者外周血中Th17細(xì)胞比例及其相關(guān)炎性細(xì)胞因子表達(dá)是否發(fā)生變化。方法:共分為三組:伴MG胸腺瘤組(TM) 35例、伴MG胸腺增生組(TH) 22例、健康對(duì)照組(HC ) 22例。使用流式細(xì)胞術(shù)實(shí)驗(yàn)檢測(cè)MG患者及健康對(duì)照外周血Th17細(xì)胞比例;采用實(shí)時(shí)定量PCR和ELISA檢測(cè)Th17細(xì)胞相關(guān)細(xì)胞因子mRNA、和蛋白表達(dá)量。結(jié)果:TM組外周血Th17細(xì)胞比例顯著高于HC (P0. 05 ) , TH組與HC組Th17細(xì)胞比例無統(tǒng)計(jì)學(xué)差異(P0.05); TM組IL-17、IL-1β,IL-6及IL-23mRNA及蛋白表達(dá)量顯著高于對(duì)照組(P0. 05 ),然而TH組與HC組IL-17、IL-6及IL-23mRNA及蛋白表達(dá)量差異無統(tǒng)計(jì)學(xué)意義(P0. 05 ),TH組IL-1 β蛋白表達(dá)量顯著高于HC組(P0. 05 )。結(jié)論:MG胸腺瘤中Th17細(xì)胞增多,相關(guān)炎性細(xì)胞因子表達(dá)增多,可能參與MG發(fā)病。第二部分:microRNA-19b在伴胸腺瘤重癥肌無力發(fā)病中表觀遺傳調(diào)控機(jī)制研究目的第一部分研究結(jié)果發(fā)現(xiàn)伴胸腺瘤MG患者的外周血Th 17細(xì)胞升高,然而Th17細(xì)胞的上游調(diào)控機(jī)制仍不清楚。胸腺間質(zhì)淋巴細(xì)胞生成素(TSLP )是Th17細(xì)胞分化發(fā)育的上游調(diào)控分子。microRNA通過轉(zhuǎn)錄后調(diào)控機(jī)制調(diào)控蛋白表達(dá)水平和生物學(xué)功能,然而microRNA在胸腺瘤細(xì)胞發(fā)育和分化一直未見報(bào)道。本研究著眼于探索伴MG胸腺瘤組織中TSLPmRNA及蛋白的表達(dá)水平以及證實(shí)microRNA-19b-TSLP-Th17細(xì)胞信號(hào)通路參與MG發(fā)病的分子機(jī)制。方法分為三組:胸腺瘤伴MG組(MG-T) 52例、單純胸腺瘤不伴MG組(NMG-T )12例和健康對(duì)照組(HC ) 11例。使用RT-PCR和Western blot測(cè)定各組胸腺瘤組織中microRNA-19b及TSLP的mRNA和蛋白表達(dá)水平;體外實(shí)驗(yàn)采用熒光素酶報(bào)告基因?qū)嶒?yàn)驗(yàn)證microRNA-19b對(duì)于TSLPmRNA的靶向調(diào)控關(guān)系;使用Spearman相關(guān)分析分析microRNA-19b與TSLPmRNA以及蛋白表達(dá)量的相關(guān)性。結(jié)果MG-T組和NMG-T組TSLPmRNA轉(zhuǎn)錄量及蛋白表達(dá)量顯著低于正常對(duì)照;同時(shí)MG-T組TSLP蛋白表達(dá)量低于NMG-T組,差異有統(tǒng)計(jì)學(xué)差異(P0. 05 );各種不同類型胸腺瘤組織TSLPmRNA轉(zhuǎn)錄量存在顯著差異,B2組TSLP、B3組TSLP低于正常胸腺。熒光素酶報(bào)告基因?qū)嶒?yàn)證實(shí)microRNA19b-5p通過與TSLP3 'UTR的相互作用靶向調(diào)控TSLPmRNA表達(dá)。胸腺瘤microRNA19b-5p成熟體、前體、初級(jí)轉(zhuǎn)錄體轉(zhuǎn)錄量顯著高于正常胸腺。microRNA19b-5p成熟體、初級(jí)轉(zhuǎn)錄體、前體與TSLP蛋白表達(dá)呈負(fù)相關(guān)。結(jié)論伴MG胸腺瘤組織中microRNA19b-5p表達(dá)上調(diào),負(fù)向靶向調(diào)控TSLPmRNA表達(dá),下調(diào)TSLP蛋白表達(dá),進(jìn)而可能影響Th17細(xì)胞的分化發(fā)育,microRNA-19b-TSLP-Th17細(xì)胞信號(hào)通路可能與MG發(fā)病相關(guān)。
[Abstract]:Text: part I: changes of Th17 cells and cytokines expression in peripheral blood of patients with thymoma myasthenia gravis objective: myasthenia gravis (MGG) is a T cell helper. The role of acetylcholine receptor antibody AchR-Abs-mediated autoimmune disease. Th17 cells in the pathogenesis of MG is still unclear. The aim of this study was to investigate the changes of Th17 cell ratio and the expression of inflammatory cytokines in peripheral blood of MG patients with thymoma. Methods: a total of 35 patients with MG thymoma, 22 patients with Thymoma with MG thymus hyperplasia and 22 patients with HC in healthy control group were divided into three groups: 35 cases with MG thymoma, 22 cases with TH with MG thymus hyperplasia and 22 cases with HC. The proportion of Th17 cells in peripheral blood of MG patients and healthy controls was detected by flow cytometry, and the expression of Th17 cytokine mRNAs and protein was detected by real-time quantitative PCR and Elisa. Results the percentage of Th17 cells was significantly higher in the group of 1: TM than that in the group of HC P0. There was no significant difference in the proportion of Th17 cells between th group and HC group (P 0.05), while the expression of IL 17 尾 IL 6 and IL 23 mRNA and protein in TM group was significantly higher than that in control group (P 0. 05). However, there was no significant difference in the expression of IL-6 and IL-23 mRNA and protein between th group and HC group (P 0. 0). The expression of IL-1 尾 protein in th group was significantly higher than that in HC group P0. 0 5. Conclusion Th17 cells and the expression of inflammatory cytokines may be involved in the pathogenesis of MG. The second part: microRNA-19b in the pathogenesis of thymoma myasthenia gravis objective the first part of the study found that the peripheral blood Th 17 cells in patients with thymoma MG increased, but the upstream regulation mechanism of Th17 cells is still unclear. TSLP is the upstream regulatory molecule of Th17 cell differentiation. MicroRNA regulates protein expression and biological function through post-transcriptional regulation mechanism. However, microRNA has not been reported in thymoma cell development and differentiation. The purpose of this study was to explore the expression level of TSLP mRNA and protein in thymoma with MG and to confirm the molecular mechanism of microRNA-19b-TSLP-Th17 cell signaling pathway involved in the pathogenesis of MG. Methods three groups were divided into three groups: 52 cases of thymoma with MG group, 12 cases of simple thymoma without MG group and 11 cases of healthy control group. The mRNA and protein expressions of microRNA-19b and TSLP in thymoma were detected by RT-PCR and Western blot, and the targeting regulation of microRNA-19b on TSLP mRNA was verified by luciferase reporter gene in vitro. Spearman correlation was used to analyze the correlation between microRNA-19b and TSLP mRNA and protein expression. Results the mRNA transcription and protein expression of TSLP in MG-T group and NMG-T group were significantly lower than those in normal control group, while the expression of TSLP protein in MG-T group was lower than that in NMG-T group (P 0). There was significant difference in TSLP mRNA transcription in different types of thymoma. TSLP in TSLP B3 group was lower than that in normal thymus. Luciferase reporter gene experiment confirmed that microRNA19b-5p regulated the expression of TSLP mRNA through the interaction with TSLP3 UTR. MicroRNA19b-5p maturation, precursor and primary transcription of thymoma were significantly higher than those of normal thymus. MicroRNA19b-5p maturation, primary transcripts and precursors were negatively correlated with the expression of TSLP protein. Conclusion the expression of microRNA19b-5p is up-regulated in MG thymoma, the expression of TSLP mRNA is regulated by negative targeting, and the expression of TSLP protein is down-regulated, which may affect the differentiation and development of Th17 cells and the signal pathway of microRNA-19b-TSLP-Th17 cells may be related to the pathogenesis of MG.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：R746.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 ;Reduction of natural regulatory T cells in thymomas accompanying myasthenia gravis and its possible association with Foxp3 and thymic stromal lymphopoietin[J];Journal of Medical Colleges of PLA;2009年01期

2 張新宇,高燕寧;PCR引物設(shè)計(jì)及軟件使用技巧[J];生物信息學(xué);2004年04期

3 吳濤,涂來慧,王志農(nóng),蔣建明,張仁琴,金海;重癥肌無力伴胸腺瘤124例臨床分析[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2003年11期

，

本文編號(hào)：2032566