本文選題：重癥肌無力 + 胸腺瘤��； 參考：《中國人民解放軍醫(yī)學院》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細胞比例及細胞因子表達量變化目的:重癥肌無力(myasthenia gravis, MG)是T細胞輔助、乙酰膽堿受體抗體(AchR-Abs)介導的神經(jīng)肌肉接頭傳遞障礙的自身免疫性疾病。Th17細胞在MG發(fā)病機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細胞比例及其相關炎性細胞因子表達是否發(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、健康對照組(HC ) 22例。使用流式細胞術實驗檢測MG患者及健康對照外周血Th17細胞比例;采用實時定量PCR和ELISA檢測Th17細胞相關細胞因子mRNA、和蛋白表達量。結果:TM組外周血Th17細胞比例顯著高于HC (P0. 05 ) , TH組與HC組Th17細胞比例無統(tǒng)計學差異(P0.05); TM組IL-17、IL-1β,IL-6及IL-23mRNA及蛋白表達量顯著高于對照組(P0. 05 ),然而TH組與HC組IL-17、IL-6及IL-23mRNA及蛋白表達量差異無統(tǒng)計學意義(P0. 05 ),TH組IL-1 β蛋白表達量顯著高于HC組(P0. 05 )。結論:MG胸腺瘤中Th17細胞增多,相關炎性細胞因子表達增多,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)控機制研究目的第一部分研究結果發(fā)現(xiàn)伴胸腺瘤MG患者的外周血Th 17細胞升高,然而Th17細胞的上游調(diào)控機制仍不清楚。胸腺間質(zhì)淋巴細胞生成素(TSLP )是Th17細胞分化發(fā)育的上游調(diào)控分子。microRNA通過轉(zhuǎn)錄后調(diào)控機制調(diào)控蛋白表達水平和生物學功能,然而microRNA在胸腺瘤細胞發(fā)育和分化一直未見報道。本研究著眼于探索伴MG胸腺瘤組織中TSLPmRNA及蛋白的表達水平以及證實microRNA-19b-TSLP-Th17細胞信號通路參與MG發(fā)病的分子機制。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例和健康對照組(HC ) 11例。使用RT-PCR和Western blot測定各組胸腺瘤組織中microRNA-19b及TSLP的mRNA和蛋白表達水平;體外實驗采用熒光素酶報告基因?qū)嶒烌炞CmicroRNA-19b對于TSLPmRNA的靶向調(diào)控關系;使用Spearman相關分析分析microRNA-19b與TSLPmRNA以及蛋白表達量的相關性。結果MG-T組和NMG-T組TSLPmRNA轉(zhuǎn)錄量及蛋白表達量顯著低于正常對照;同時MG-T組TSLP蛋白表達量低于NMG-T組,差異有統(tǒng)計學差異(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。熒光素酶報告基因?qū)嶒炞C實microRNA19b-5p通過與TSLP3 'UTR的相互作用靶向調(diào)控TSLPmRNA表達。胸腺瘤microRNA19b-5p成熟體、前體、初級轉(zhuǎn)錄體轉(zhuǎn)錄量顯著高于正常胸腺。microRNA19b-5p成熟體、初級轉(zhuǎn)錄體、前體與TSLP蛋白表達呈負相關。結論伴MG胸腺瘤組織中microRNA19b-5p表達上調(diào),負向靶向調(diào)控TSLPmRNA表達,下調(diào)TSLP蛋白表達,進而可能影響Th17細胞的分化發(fā)育,microRNA-19b-TSLP-Th17細胞信號通路可能與MG發(fā)病相關。
[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.
【學位授予單位】：中國人民解放軍醫(yī)學院
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R746.1

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本文編號：2032566