强直性脊柱炎（AS）患者外周血单个核细胞中miRNAs的表达与血清细胞因子相关性研究

发布时间：2018-06-23 14:32

本文选题：强直性脊柱炎 + miR-155　；参考：《安徽医科大学》2015年硕士论文

【摘要】：背景微小RNA(mic RNA)是一类染色体上高度保守的非编码单链RNA,广泛存在于真核生物中,它通过与靶基因特异性结合,引起靶基因m RNA的降解或干扰蛋白质的翻译过程,并可在特定条件下上调靶基因的蛋白表达和信使RNA的水平。mi RNA参与了生物体多种生理和病理过程,并在固有免疫、适应性免疫及炎症因子的信号转导过程中发挥着关键性的调节作用。强直性脊柱炎(ankylosing spondylitis,AS)是一种类风湿因子阴性的慢性炎症性自身免疫性疾病,以侵犯中轴骨骼为主,时常累及外周关节、肌腱韧带附着点及其它软骨组织,引起脊柱强直和纤维化。AS的病因及发病机制至今仍不明确,可能与遗传、免疫、慢性感染、遗传与环境的交互作用等有关。细胞因子在免疫细胞的生长、分化及免疫调节中发挥信号传递作用。本课题组前期发现了多种细胞因子,如IL-6、IL-12、IL-17在AS中的异常表达,本研究拟通过检测AS患者部分mi RNA的表达水平并探讨与以上细胞因子的相关性,为强直性脊柱炎病因及治疗提供线索和依据。目的通过检测AS患者外周单个核细胞中mi R-155、mi R-16、mi R-31、mi R-181a表达水平,并分析mi R-155、mi R-16、mi R-31、mi R-181a与血清细胞因子、临床指标、疾病活动度的相关性,探讨mi R-155、mi R-16、mi R-31、mi R-181a在AS发生和发展中所起到的作用及意义。方法采用荧光定量PCR检测40名AS患者和40名健康对照组外周单个核细胞中mi R-155、mi R-16、mi R-31、mi R-181a表达水平。AS患者来源于安徽医科大学第一附属医院风湿免疫科,根据强直性脊柱炎活动指数将AS患者分为活动组和稳定组;选取同期合肥市中心血站健康献血者作为对照组。采用酶联免疫吸附法(ELISA),检测AS患者细胞因子IL-6、IL-12、IL-17、IL-10,TNF-a和健康对照血清TNF-a,IL-10的水平,分析AS组与健康对照组间TNF-a,IL-10的水平。进一步分析病例组mi R-155、mi R-16、mi R-31、mi R-181a表达水平与细胞因子、疾病活动度、ESR、CRP等临床指标的相关性。结果本研究发现AS患者外周单个核细胞中mi R-31的表达显著高于健康对照组(Z=-4.007,p0.001),而mi R-155,mi R-16,mi R-181a外周血单个核细胞表达在强直性脊柱炎和健康对照组之间无统计学差异(Z=-1.095,P=0.274;Z=-1.555,P=0.120;Z=-1.720,P=0.086)。病情稳定组与活动组中mi R-155,mi R-16,mi R-181a,mir-31的表达无差异(Z=-0.525,P=0.607;Z=-0.818,P=0.421;Z=-0.812,P=0.434;Z=-0.66,P=0.517)。同时,mi R-155,mi R-16,mi R-181a,mi R-31的表达与各细胞因子IL-6、IL-12、IL-17、IL-10,TNF-a均无相关性(P均0.05)。mi R-155,mi R-16,mi R-181a,mi R-31的表达与疾病活动指数、ESR、CRP之间均无相关性(p均0.05)。但IL-6、IL-12与疾病活动指数具有相关性(p0.05)。结论AS患者外周血单个核细胞中mi R-31表达上调,提示可能在AS的发病中起到重要的作用。
[Abstract]:Background microRNAs are a class of highly conserved non-coding single-stranded RNAs on chromosomes, which are widely found in eukaryotes. By specifically binding with target genes, mic RNAs cause the degradation of target genes and interfere with the process of protein translation. And can up-regulate the protein expression of target gene and the level of messenger RNA under certain conditions. Mi RNA is involved in many physiological and pathological processes of organism and innate immunity. Adaptive immunity and the signal transduction of inflammatory factors play a key role in regulation. Ankylosing spondylitis as (as) is a chronic inflammatory autoimmune disease with negative rheumatoid factor. The etiology and pathogenesis of spinal ankylosis and fibrosis as are still unclear, which may be related to heredity, immunity, chronic infection, interaction between heredity and environment, etc. Cytokines play a signalling role in the growth, differentiation and immunomodulation of immune cells. Several cytokines, such as the abnormal expression of IL-6, IL-12, IL-17, were found in the early stage of the study. This study was intended to detect the expression level of partial mi RNA in patients with as and to explore the correlation with the above cytokines. To provide clues and evidence for the etiology and treatment of ankylosing spondylitis. Objective to detect the expression level of mi R-155n mi R-16tmmi R-31a in peripheral mononuclear cells (PBMC) of as patients, and to analyze the correlation between the expression of mi R-155Mi R-16mmi R-31mmiR-181a and serum cytokines, clinical indexes and disease activity. This paper discusses the role and significance of mi R-155U mi R-16C mi R-31M R-181a in the occurrence and development of as. Methods fluorescence quantitative PCR was used to detect the expression of mi R-155N mi R-16M R-31mmi R-181a in peripheral mononuclear cells of 40 patients with as and 40 healthy controls. The patients with as were from the Department of Rheumatology and Immunology of the first affiliated Hospital of Anhui Medical University. According to the activity index of ankylosing spondylitis, patients with as were divided into active group and stable group. Enzyme linked immunosorbent assay (Elisa) was used to detect the levels of the cytokine IL-6, IL-12, IL-17, IL-10, and the serum levels of TNF-a, IL-10 in healthy controls, and the levels of TNF-a IL-10 between as group and healthy control group were analyzed by enzyme-linked immunosorbent assay (Elisa). Further analysis was made on the correlation between the expression of miR-155nmi R-16mR-31mil R-181a and cytokines, disease activity and ESR-CRP, and so on. Results the expression of miR-31 in peripheral mononuclear cells in patients with as was significantly higher than that in healthy controls (ZF-4.007p0.001), while the expression of MIR-155mR-16mR-181a in peripheral blood mononuclear cells was not significantly different between the patients with ankylosing spondylitis and the healthy controls (Z-1.095P0.274Z-1.555P0. 120-1.720P0.086). The expression of miR-31 in peripheral mononuclear cells in patients with as was significantly higher than that in healthy controls (Z-4.007p0.001), but there was no significant difference in the expression of MIR-181a between ankylosing spondylitis and healthy controls. There was no difference in the expression of mi R-155nmi R-16nmi R-181a mir-31 between the stable group and the active group (ZAM-0.525, P ~ 0.607, P ~ (0.607), P ~ (0.421), P ~ (0.421), P ~ (0.434), P ~ (0.434), Z ~ (-0.66), P _ (0.517). At the same time, there was no correlation between the expression of R-155Mi R-181ammi R-31 and the cytokines IL-6, IL-12, IL-17, IL-10, TNF-a (P 0.05) .mi R-155mi R-165mi R-181a mi R-31 and disease activity index (ESRCRP) (p < 0.05). But IL-6 and IL-12 were correlated with disease activity index (p0.05). Conclusion the expression of miR-31 in peripheral blood mononuclear cells is up-regulated in patients with as, suggesting that it may play an important role in the pathogenesis of as.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.23

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