miR-155与自身免疫性疾病研究进展

微小RNA(microRNA,miRNA)介导的基因表达对于维持正常的细胞周期、分化、增殖、凋亡以及维持免疫系统的稳定性方面发挥着非常重要作用.研究发现miR-155与炎症、自身免疫性疾病密切相关,负性调节炎症和固有免疫反应,可作为疾病治疗的潜在靶点.本文就miR-155与自身免疫性疾病的研究进展进行综述.     

miR-146a调控血小板免疫活化功能

目的探讨miR-146a对血小板免疫活化功能的影响。方法流式细胞术检测冠心病(n=31)及健康成年人(n=35)全血中血小板PAC-1、CD62-P阳性率,q-PCR及光比浊法分别检测血浆中miR-146a及血小板聚集率;培养K562细胞,佛波酯(PMA)诱导分化为巨核细胞后,分别转染miR-146a mimics、inhibitor及其阴性对照,Western blot检测转染后细胞中TLR-4、PAC-1及CD62-P表达水平。结果冠心病患者miR-146a、血小板聚集率及PAC-1、CD62-P阳性率较对照组升高(P0.05);转染miR-146a mimics后,TLR-4、PAC-1和CD62-P表达水平升高(P0.05)。结论miR-146a可能依赖血小板TLR-4信号通路,介导血小板免疫活化进而促进血小板聚集,加速血栓形成。     

miRNA 表达与免疫细胞因子相关性在原发性肝癌早期诊断和预后中的作用

目的:检测miR-146a、miR-224、miR-34c、miR-200a、miR-148b、miR-375 六种miRNAs 在原发性肝癌中的表达变化及其与HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc 和IL-12、IL-4、IL-6、IL-10、IFN-γ、TNF-α等炎症因子表达的相关性,以验证循环miRNA 是否可作为理想的血源性新型生物标志物用于原发性肝癌的早期检测。方法:收集肝炎、肝硬化患者及健康对照组静脉血,并收集原发性肝癌患者癌组织和癌旁组织。提取总RNA 后通过实时定量PCR 检测并比较各组miRNA 的相对表达水平,同时检测miRNA 表达水平变化与血清肿瘤标志物AFP、CEA、CA19-9、CA125 表达的关系;并检测miRNAs 表达水平变化与HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc 和炎症因子IL-12、IL-4、IL-6、IL-10、IFN-γ、TNF-α表达相关性。结果:相对于健康组,miR-34c、miR-224、miR-146a 在PHC 组血清和组织中表达显著上调;miR-200a、miR-148b、miR-375 在PHC 组血清和组织中表达显著下调,差异具有统计学意义。HBsAg 与血清miR-375 和miR-146a 存在回归关系,miR-375 随HBsAg 表达水平升高而降低,而miR-146a 随HBsAg 表达升高而升高。IFN-γ与miR-146a 存在回归关系, miR-146a 随IFN-γ表达水平降低而升高,miR-375 和miR-146a 诊断能力大于CA19-9 和AFP。结论:miR-146a、miR-224、miR-34c、miR-200a、miR-148b、miR-375 在原发性肝癌血清和组织中存在表达差异,其中miR-375 和miR-146a 诊断能力优于AFP 和CA19-9,血清miR-375 和miR-146a 可能成为新的肝癌早期诊断标志。     

MiR-146a在免疫反应中的调节作用

microRNA是一类非编码的小分子RNA,通过在转录后水平抑制蛋白的表达,在许多生理和病理进程中发挥重要作用。MiR-146a在固有免疫调节、炎症反应以及抗病毒中发挥重要作用。本文就最近对miR-146a在固有免疫、炎症应答、病毒感染和人类疾病等方面的研究进行了综述。研究显示,可能可以通过对miR-146a表达的调控来治疗一些人类疾病。     

固有免疫功能相关的microRNAs 在布鲁氏菌病患者中的表达情况

目的:研究与固有免疫相关的14 种microRNAs 在布病患者中的表达情况。方法:采用实时定量PCR 方法,测定布病患者和健康志愿者血清中microRNAs 的表达水平。结果:在检测的14 种microRNAs 中,13 种在布病患者中表达下调,其中布病患者和健康志愿者之间的Ct值差异>1 的microRNAs 共有9 种,并且miR-122、miR-145a、miR-155、miR-301a 和miR-146a 这5 种microRNAs 在布病患者血清中的表达水平明显降低(P<0.01)。结论:布病患者中miR-122、miR-145a、miR-155、miR-301a 和miR-146a 的表达被明显抑制,在布病诊断中具有潜在的应用价值。     

微小RNA-146a在自身免疫性疾病中的研究进展

微小RNA(microRNA,miRNA)是一类大小在21~25个核苷酸的单链内源性非编码RNA,通过转录后的抑制广泛参与不同细胞生物学过程。MiR-146a是miRNA家族中成员之一,参与细胞增殖、分化、凋亡等生物学过程,同时也在恶性肿瘤和炎症疾病的进展中发挥调节作用。进一步研究发现,miR-146a在不同类型的免疫...     

miR-146a通过靶向调控TLR4减轻脊髓缺血再灌注大鼠损伤

目的 检测miR-146a在脊髓缺血再灌注(IR)大鼠脊髓组织的表达情况,探索miR-146a对IR大鼠炎症蛋白表达和神经功能影响的潜在机制.方法 SD大鼠通过主动脉弓夹闭14min建立IR模型,通过鞘内注射miR-146a的mimics质粒构建miR-146a过表达模型,随机分为Sham组(假手术组),IR组(缺血组...     

miR-146b 免疫学研究进展

miR-146家族包括两个成员miR-146a和miR-146b,与miR-146a不同,miR-146b在免疫应答方面的生物学信息很少;本文讨论了miR-146b的产生、作用机制、信号通路以及在相应疾病中的作用,为miR-146b的进一步研究提供思考和借鉴。     

MicroRNA-21在自身免疫性疾病中的调节作用与机制

MicroRNAs(miRNAs)是一类具有调控功能的小分子非编码RNA,其介导的基因表达对于维持正常细胞的增殖、分化、凋亡以及维持免疫系统的稳定有着重要作用.大量研究结果表明,miR-21通过Toll样受体(TLR)信号途径参与了固有免疫应答和炎症调节,而且它还可以通过多种方式对免疫细胞的生长、分化等产生影响.近年来,关于miR-21在自身免疫性疾病中的作用及其机制研究备受关注,本文对miR-21调节免疫反应及其对几种常见自身免疫性疾病的调节作用进行了综述.     

miR-146a在哮喘患儿痰液的表达及对哮喘小鼠气道炎症的影响

目的探讨miR-146a在哮喘患儿痰液中的表达及其对气道炎症的作用。方法应用real-time PCR方法检测哮喘患儿和健康儿童痰液,以及卵清蛋白诱导的哮喘小鼠和正常小鼠肺组织中miR-146a的表达;化学合成miR-146a模拟物与阴性对照,并分别滴鼻至哮喘小鼠,HE染色验证哮喘小鼠模型的成功建立,免疫荧光方法和Western blot方法检测各组小鼠肺组织白介素-1β(IL-1β)与白介素-4(IL-4)的蛋白表达。结果与对照组相比,哮喘患儿痰液及哮喘小鼠肺组织中miR-146a表达显著升高(P0.01);HE染色显示哮喘模型组小鼠炎症细胞浸润明显高于正常对照组,哮喘小鼠模型成功建立。miR-146a模拟物滴入后,哮喘小鼠肺组织IL-1β与IL-4的蛋白表达显著降低(P0.01)。结论 miR-146a在支气管哮喘患儿及支气管哮喘小鼠模型中高表达,miR-146a模拟物能够抑制哮喘小鼠的气道炎症。     

中国广西人群miR-146a、miR-149 基因遗传多态性的研究

目的：研究广西人群miR-146a C>G (rs2910164）、miR-149 T>C（rs2292832）等位基因及基因型频率分布,分析其与不同民族种族间的差异性。方法：采用单碱基延伸技术和DNA测序技术对303例广西人群中miR-146a C>G和miR-149 T>C基因单核苷酸多态性(SNP)位点进行分型检测,并与人类基因组计划 (Hapmap) 数据库中公布的非洲人、欧洲人、日本人和中国北京人群的SNP分型数据比较。结果：miR-146a C>G、miR-149 T>C等位基因和基因型频率在广西男女人群之间分布均无差异性（P均>0.05）。广西人群miR-146a C>G和miR-149 T>C基因型及等位基因频率与非洲人、欧洲人、中国北京人比较有统计学意义（P均<0.05）。结论：广西人群miR-146a C>G和miR-149 T>C存在基因多态性,与其他种族人群比较有差异性,这种差异性对人类遗传病的研究可能会起到重要作用。     

MicroRNA-146a promotes IgE class switch in B cells via upregulating 14-3-3σ expression

B cells play a critical role in immune responses both in physiological and pathological conditions, and microRNAs have been shown to play important roles in regulating B cell proliferation and function. MiR-146a has been shown to modulate T cell immunity, but its function in regulating B cell response remains partially understood. Our previous studies indicated that germinal center (GC) B cells are significantly expanded in miR-146a-overexpressing (TG) mice. In this study, we further characterized the roles of miR-146a in regulating humoral immune responses to specific antigens. We found that the production of IgE antibody were significantly elevated in TG mice, while the antibody affinity maturation of IgM and IgG were similar between TG mice and the normal controls. We further found higher IgE antibody levels in TG B cell culture supernatant than that in normal controls. A global protein expression comparison of B cells from TG mice and the normal controls through TMT proteomic assay showed that 14-3-3σ, a key factor of immunoglobulin class switch DNA recombination (CSR) in B cells, was highly up-regulated in B cells with overexpression of miR-146a, while Smad4, the target of miR-146a, was decreased. Using an asthma model induced by OVA immunization, we further confirmed the increased level of OVA specific IgE antibodies in TG mice. These results demonstrate that miR-146a enhances class switch and secretion of IgE in B cells by upregulating 14-3-3σ expression, and suggest that miR-146a may be a potential target for asthma therapy.     

系统性红斑狼疮患者中miR-146a表达缺陷与I型干扰素通路过度活化相关

Ι型干扰素(IFN)通路过度活化与固有免疫反应异常在系统性红斑狼疮(SLE)发病中起重要作用。miR-146a作为一个负调控因子,在固有免疫应答中发挥重要作用。为探索miR-146a在SLE发病中的作用,通过实时荧光定量聚合酶链反应技术检测18例SLE患者和11例正常对照miR-146a的表达,发现同正常对照组相比,miR-146a表达水平在SLE患者中明显降低,SLE活动组中下降则更加明显;同时检测miR-146a的靶基因IRAK1和TRAF6 mRNA表达,发现在SLE患者中其水平显著高于正常对照组;进一步分析miR-146a与IFN诱导基因表达之间的关系,结果显示两者之间存在负相关,体外实验也证实miR-146a能够负向调节I型干扰素通路。综上所述,我们的研究结果提示miR-146a可能作为一个新的生物标志物,SLE患者中miR-146a表达水平下降,导致I型干扰素通路负调控作用被削弱,从而参与疾病的发生发展。     

MiR-146a modulates macrophage polarization in systemic juvenile idiopathic arthritis by targeting INHBA

Monocytes from patients with systemic juvenile idiopathic arthritis (SJIA) have both features of classical activated M1 and alternatively activated M2 macrophages. An increasing number of studies have indicated that microRNAs (miRNAs) are critical regulators of monocyte polarization. Here, we focused on miR-146a expression in SJIA and investigated the function of miR-146a in monocyte polarization. We found that miR-146a expression was highly up-regulated in SJIA monocytes and correlated with the systemic features. miR-146a was expressed at a higher level in monocytes polarized with M2 conditions than those polarized with M1 conditions. miR-146a overexpression significantly decreased the production of M1 phenotype markers such as IL-6, IL-12, TNF-α, CD86 and iNOS in M1 macrophages, but increased the production of M2 marker genes such as Arg1, CCL17, CCL22 and CD206 in M2 macrophages. Conversely, knockdown of miR-146a promoted M1 macrophage polarization but diminished M2 macrophage polarization. We subsequently demonstrated that miR-146a targeted the 3′-untranslated region (UTR) of INHBA to inhibit its expression. Additionally, INHBA overexpression rescued the reduced IL-6, IL-12, and TNF-α levels induced by miR-146a overexpression in M1 macrophages, and rescued the increased Arg1, CCL17, and CCL22 levels induced by miR-146a overexpression in M2 macrophages. Similarly, the effects of miR-146a inhibition in monocyte polarization were all partly reversed by INHBA inhibition. Taken together, the data suggest that miR-146a serves as a molecular regulator in monocyte polarization and might play an important role in monocytes from patients with SJIA.     

The occurrence of cervical cancer in Uygur women in Xinjiang Uygur Autonomous Region is correlated to microRNA-146a and ethnic factor

Aims: The present study is to investigate the effect of microRNA-146a (miR-146a) and ethnic factor in the occurrence of cervical cancer in Uygur women in Xinjiang Uygur Autonomous Region. Methods: A total of 620 pieces of cervical tissues were obtained between September 2010 and September 2013, including 208 cases of cervicitis, 207 cases of cervical intraepithelial neoplasia, and 205 cases of cervical cancer. The relative expression of miR-146a in tissues was measured using quantitative real-time polymerase chain reaction. Polymerase chain reaction - restriction fragment length polymorphism was used to determine the genotypes of miR-146a (rs2910164). Differences between two groups and multiple groups were compared using t-test and one-factor analysis of variance, respectively. Comparison of genotype compositions and genetic balance examinations were performed using χ² test. Results: Uygur women had earlier age of marriage, more times of pregnancy, and more childbirths than Han women. The miR-146a (rs2910164) genotype composition was significantly different between Uygur and Han, with the ratio of GG genotype in Uygur being higher than that in Han. Logistic regression analysis showed that miR-146a (rs2910164) genotypes were significantly correlated to ethnic factor and tumor sizes. The expression of miR-146a was elevated in cervical intraepithelial neoplasia and cervical cancer, especially for Uygur women, with the GG genotype being the most highly expressed. Conclusions: The miR-146a (rs2910164) polymorphism is significantly correlated to ethnic factor and tumor diameters. miR-146a has differential expression in cervical tissues. Allele G of miR-146a (rs2910164) is related to the high expression of miR-146a, and the progression of cervical cancer.     

miR-146b-5p promotes VSMC proliferation and migration

Vascular smooth muscle cells (VSMCs) play pivotal roles in the development of vascular diseases. While microRNAs are important in vascular pathologies, a few is known about their functional roles in VSMC phenotypes. We profiled microRNA expression in PDGF-BB treated VSMCs and found microRNA-146b-5p (miR-146b-5p) was upregulated. Inhibition of miR-146b-5p blocked in response to PDGF while reducing VSMC proliferation and migration. These studies implicate miR-146b-5p as necessary for PDGF-induced VSMC phenotype transition. Downstream miR-146b-5p targets modulating VSMC phenotypes will be further identified. Our study will help to understand the role of VSMCs in the pathology of vascular diseases.     

微小RNA-146a 上调cyclin D1 表达诱导大鼠血管平滑肌细胞的增殖

目的:采用基因芯片技术研究小分子RNA-146a(miR-146a)促进血管平滑肌细胞增殖的作用靶点并进行验证。方法:原代培养大鼠血管平滑肌细胞,分成mimics 组、inhibitor 组、control 组、sham 组,分别体外转染miR-146a mimics(50nmol/ L)、miR-146a inhibitor(50 nmol/ L)、miR-146a 错义链(50 nmol/ L)、PBS,Real time PCR 测定转染后miR-146a 水平,CCK8法检测转染后血管平滑肌细胞增殖情况。采用基因芯片检测inhibitor 组和control 组基因表达谱,通过生物信息学技术筛选出差异基因和调控的信号通路。对筛选出来的信号通路用Real time PCR 和Western blot 进行验证。结果:转染48 h后,inhibitor 组血管平滑肌细胞的miR-146a 水平明显低于control 组和sham 组(P<0.01),inhibitor 组血管平滑肌细胞的OD 值明显低于control 组、sham 组(P<0.05)。通过对基因表达谱分析发现,p53 信号通路被miR-146a 上调。用Real time PCR 和Western blot进行检测发现,p53 信号通路中关键分子p53、caspase3、PTEN 的mRNA 和蛋白水平无明显变化(P>0.05),而mimics 组VSMC中cyclin D1 的mRNA 和蛋白水平增加(与sham 相比,P<0.05),inhibitor 组VSMC 中cyclin D1 的mRNA 和蛋白水平下降(与sham相比,P<0.05)。结论: miR-146a 可能通过上调细胞周期蛋白cyclin D1 的表达促进大鼠血管平滑肌细胞的增殖。     

Regulatory effects of miR-155 and miR-146a on repolarization and inflammatory cytokine secretion in human alveolar macrophages in vitro

Macrophages play an important role in inflammatory responses; however, miRNA-mediated repolarization of macrophages is essential for fulfilling this function. To clarify a series of changes at the RNA level in alveolar macrophages under normal and inflammatory conditions, bronchial alveolar lavage liquid (BALF) was collected from healthy volunteers or patients with pneumonia. This approach, which differs from that used in previously, provides more accurate information about the states of macrophages in different lung microenvironments. In this study, the density plots of macrophage subtypes (M1 and M2) in the BALF of healthy volunteers differed from that of the patients with pneumonia. The M2 subtype dominated in healthy volunteers and was rapidly repolarized to M1 in response to miRNA-mediated gene regulation. Differential miRNA expression in the two macrophage subtypes revealed lower expression of miR-155 and MIR-146a in patients with pneumonia compared with healthy volunteers; this may be related to inflammation and the use of anti-inflammatory drugs. We also found increased TNF-α and IL-6 expression at the RNA level, while macrophage galactose-type C-type lectin 1 (MGL-1) expression decreased with downregulation of miR-155 and miR-146a expression. These results indicate that the gene regulation mediated by miR-155 and miR-146a contributes to human alveolar macrophage phenotype repolarization, thus leading to an early switch from pro-inflammatory to anti-inflammatory cytokine production.