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腦膠質(zhì)瘤患者外周血Th17、Th22細(xì)胞的表達(dá)及意義

發(fā)布時(shí)間:2018-10-13 15:52
【摘要】:研究背景膠質(zhì)瘤是顱內(nèi)最常見的原發(fā)性惡性腫瘤,近年來其發(fā)病率有逐年上升的趨勢(shì),由于腫瘤侵犯腦實(shí)質(zhì),侵襲、轉(zhuǎn)移能力強(qiáng),復(fù)發(fā)率高,5年生存率極低。目前,國內(nèi)外治療膠質(zhì)瘤的措施仍是以手術(shù)治療為主,輔以放、化療及其他綜合治療。以往由于血腦屏障的存在,腦一直被看做是一個(gè)免疫豁免器官,但現(xiàn)在隨著神經(jīng)系統(tǒng)免疫學(xué)研究的深入,人們發(fā)現(xiàn)免疫細(xì)胞可以通過血腦屏障,T細(xì)胞可在腦內(nèi)活化,這為腦膠質(zhì)瘤的免疫治療提供了基礎(chǔ)。Th17、Th22細(xì)胞是新近發(fā)現(xiàn)的CD4+T淋巴細(xì)胞群,其中Th17細(xì)胞主要分泌IL-17因子,RORC是其分化的特異性轉(zhuǎn)錄調(diào)控因子;Th22細(xì)胞主要分泌IL-22因子,AHR是其分化的特異性轉(zhuǎn)錄調(diào)控因子。最近研究發(fā)現(xiàn),Th17、Th22細(xì)胞在自身免疫性疾病、感染性疾病、腫瘤、血液病中均有異常表達(dá),二者參與上述疾病的發(fā)生與進(jìn)展過程,但二者在腦膠質(zhì)瘤中的表達(dá)及其作用的研究相對(duì)較少。研究目的通過研究腦膠質(zhì)瘤患者外周血中Th17、Th22細(xì)胞比例的變化及二者的相關(guān)關(guān)系,檢測(cè)外周血單個(gè)核細(xì)胞RORC mRNA及AHR mRNA的表達(dá)及血清中IL-22的濃度,旨在探究Th17、Th22細(xì)胞及其相關(guān)因子在腦膠質(zhì)瘤中的表達(dá)情況,初步探討其在腦膠質(zhì)瘤發(fā)生發(fā)展中的作用。研究方法選取2014年7月至2015年8月山東大學(xué)第二醫(yī)院神經(jīng)外科接受治療的34例膠質(zhì)瘤患者為病例組,并按其病理類型分組,其中低級(jí)別組(WHO Ⅰ-Ⅱ)共17例,高級(jí)別組(WHO Ⅲ-Ⅳ)共17例,同期24例健康體檢者為對(duì)照組,均收集其晨起空腹肘靜脈全血。1.應(yīng)用流式細(xì)胞術(shù)檢測(cè)外周血中Th17、Th22細(xì)胞的表達(dá)比例,其中Thl7定義為CD4+IFN-γ-IL-17+, Th22定義為CD4+IFN-γ- IL-17- IL-22+,并分析二者的比例變化及其相關(guān)關(guān)系。2.采用Ficoll密度梯度離心法分離外周血單個(gè)核細(xì)胞,用Trizol法提取細(xì)胞中的總RNA,逆轉(zhuǎn)錄后用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)技術(shù)檢測(cè)外周血單個(gè)核細(xì)胞中RORC mRNA及AHRmRNA的表達(dá)水平。3.采用酶聯(lián)免疫吸附分析(Enzyme linked immunosorbent assay, ELISA)檢測(cè)血漿中Th22細(xì)胞相關(guān)因子IL-22的水平。研究結(jié)果1.與健康對(duì)照組相比,腦膠質(zhì)瘤患者組外周血Th17細(xì)胞比例(占淋巴細(xì)胞)有升高趨勢(shì),但兩組間差異無統(tǒng)計(jì)學(xué)意義(P0.05),且高級(jí)別組(WHOⅢ-Ⅳ級(jí))與低級(jí)別組(WHO Ⅰ-Ⅱ級(jí))之間的差異無統(tǒng)計(jì)學(xué)意義(P0.05)。2.與健康對(duì)照組相比,腦膠質(zhì)瘤患者組外周血Th22細(xì)胞比例(占淋巴細(xì)胞)明顯升高(P0.05),且高級(jí)別組升高程度明顯高于低級(jí)別組(P0.05)。腦膠質(zhì)瘤患者外周血Th17與Th22細(xì)胞的表達(dá)呈正相關(guān)關(guān)系(r=0.40,P=0.03),健康對(duì)照組二者表達(dá)無相關(guān)關(guān)系(P0.05)。3.與健康對(duì)照組比較,腦膠質(zhì)瘤患者外周血單個(gè)核細(xì)胞RORC mRNA及AHR mRNA的表達(dá)水平升高不明顯(P0.05;P0.05);且高級(jí)別組與低級(jí)別組相比,這兩種因子的表達(dá)差異不明顯(P0.05;P0.05)。4.腦膠質(zhì)瘤患者血漿IL-22水平比健康對(duì)照組明顯升高(P0.05),但高級(jí)別組與低級(jí)別組組間差異不明顯(P0.05)。研究結(jié)論1.腦膠質(zhì)瘤患者外周血中存在Th22細(xì)胞及其相關(guān)因子IL-22的異常升高,外周血Th22細(xì)胞及IL-22的變化或可作為監(jiān)測(cè)腦膠質(zhì)瘤發(fā)病及進(jìn)展的指標(biāo)。2.腦膠質(zhì)瘤患者Th17與Th22細(xì)胞明顯相關(guān),二者可能共同參與腦膠質(zhì)瘤的發(fā)生與發(fā)展過程,并有可能發(fā)揮協(xié)同作用。
[Abstract]:Background glioma is the most common primary malignant tumor in the brain. In recent years, the incidence of glioma has been increasing year by year. Because of tumor invasion, invasion and metastasis ability is strong, the recurrence rate is high, and the 5-year survival rate is very low. At present, the treatment of glioma at home and abroad is still mainly operated by surgery, supplemented by radiotherapy, chemotherapy and other comprehensive treatment. In the past due to the existence of blood-brain barrier, brain has been regarded as an immune-immune organ, but with the in-depth study of nervous system immunology, it has been found that immune cells can be activated in the brain through the blood-brain barrier and T-cells, which provides the basis for the immunotherapy of glioma. Th17, Th22 cells are newly discovered CD4 + T lymphocyte populations, among which Th17 cells secrete IL-17 factor, RORC is the specific transcription regulation factor of its differentiation; Th22 cells secrete IL-22 factor mainly, and AHR is the specific transcription regulation factor of its differentiation. It has been found that Th17 and Th22 cells have abnormal expression in autoimmune diseases, infectious diseases, tumors and blood diseases, both of which are involved in the occurrence and progression of these diseases, but their expression and their role in gliomas are relatively few. Objective To investigate the changes of Th17 and Th22 cell ratios in peripheral blood of glioma patients and their correlation, and to detect the expression of RORC mRNA and AHR mRNA in peripheral blood mononuclear cells and the concentration of IL-22 in serum. The expression of Th22 cells and their associated factors in gliomas was studied, and the role of Th22 cells in the development of glioma was discussed. Methods 34 glioma patients treated with neurosurgery from July 2014 to August 2015 were selected as case group and grouped according to their pathological types. Among them, there were 17 cases in the lower group (WHO I-鈪,

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