类风湿性关节炎患者IL-6、IL-18和CRP的水平变化及意义

目的：研究类风湿性关节炎（RA）患者血清白细胞介素-6（IL-6）、白细胞介素-18（IL-18）和C-反应蛋白（CRP）水平的变化及其临床意义。方法：收集84例RA患者,以70例健康体检者作对照。采用双抗体夹心酶联免疫吸附法测定血清IL-6、IL-18和免疫荧光法测定CRP的水平,并测定血小板计数（Plt）、血沉（ESR）、类风湿因子（RF）。结果：RA患者的血清Plt、ESR、RF、IL-6、IL-18和CRP的含量明显高于健康对照组（P〈0.01）。RA患者活动期上述指标含量（除RF外）均显著高于稳定期（P〈0.01）,Plt升高RA患者组与Plt正常组相比,RF、ESR、IL-6、IL-18和CRP水平均有明显统计学差异（P〈0.05）。结论：IL-6、IL-18和CRP在RA患者的疾病发展过程中发挥着重要作用,它们的水平变化与RA患者病情有关,联合动态监测有助于临床观察RA患者的病情变化和治疗效果。     

Tolicizumab与风湿免疫性疾病

IL-6在类风湿关节炎(RA)、幼年特发性全身性关节炎(sJIA)、Castleman病、系统性红斑狼疮(SLE)等多种风湿免疫性疾病发生、进展过程中均有过量表达,且发挥关键作用.Tolicizumab是第一个人源性的IL-6R抗体,通过特异性识别结合IL-6R而阻断IL-6生物学活性,发挥抑制上述疾病炎症反应的作用.大量研究结果证实,tocilizumab不仅能明显缓解RA的临床症状,而且能防止关节继续破坏.此外,tocilizumab在治疗全身性和关节型sJIA、Castleman病、SLE、克罗恩病等风湿免疫性疾病也获得了巨大的成功.     

Tolicizumab研究进展

类风湿关节炎（RA）、幼年特发性全身性关节炎（sJIA）、Castleman病等多种炎性疾病发生、进展过程中均有IL-6的过量表达,且发挥关键作用。Tolicizumab是第一个人源性的IL-6R抗体,通过特异性识别结合IL-6R而阻断IL-6生物学活性,发挥抑制上述疾病炎症反应的作用。大量研究结果证实。Tocilizumab不仅能明显缓解RA的临床症状,而且能防止关节继续被破坏。此外,Tocilizumab在治疗全身性和关节型sJIA、Castleman病等疾病也获得了巨大的成功。     

类风湿关节炎患者血清IL-6、TNF-α与性激素水平的变化

类风湿关节炎(RA)是一种自身免疫性疾病，其发病机制与多种因素有关。其中细胞因子与性激素分泌异常在RA发病中起到重要作用。我们对RA患者血清白细胞介素6(IL-6)、肿瘤坏死因子α(TNF—α)以及雌二醇(E2)、睾酮(TEST)的水平进行了检测，以探讨其在RA的发生、发展中的作用。     

类风湿性关节炎滑膜成纤维细胞骨保护素和骨形态发生蛋白-2的表达及干预

目的 探讨甲氨蝶呤（MTX）及白细胞介素-6受体（IL-6R）抗体对类风湿关节炎（RA）滑膜成纤维细胞增殖的干预及对骨保护素（OPG）、骨形态发生蛋白-2（BMP-2）的影响.方法 获取RA患者滑膜组织,消化并传代培养成纤维细胞.CCK-8法检测IL-6R抗体、甲氨蝶呤（MTX）对RA滑膜成纤维细胞活性影响;荧光定量（qRT）-PCR检测OPG和BMP-2表达.结果 与空白组相比,MTX组、IL-6R抗体组成纤维细胞的活性受到抑制（F=29.30,34.22,P＜0.05）,MTX联合IL-6R抗体组成纤维细胞的活性显著受到抑制（F=52.04,P ＜0.01）.qRT-PCR显示与空白组相比,IL-6R抗体组、MTX联合IL-6R抗体组OPG、BMP-2的表达均上升（P＜0.05）.与MTK组相比,IL-6R抗体组OPG、BMP-2的表达增加（P＜0.05）.结论 IL-6R抗体单独或联合MTK使用均能明显抑制RA滑膜成纤维细胞的活性,与MTX相比,IL-6R抗体能升高OPG、BMP-2的表达.本研究为应用IL-6R抗体预防和治疗RA关节破坏提供实验依据.     

抗突变型瓜氨酸波形蛋白抗体在诊断类风湿性关节炎中的意义

目的 探讨抗突变型瓜氨酸波形蛋白抗体(anti-mutated citrullinated vimentin antibody,Anti-MCV)在诊断类风湿性关节炎(rheumatoid arthritis,RA)中的价值.方法 选取确诊的RA患者51例(RA组)、非RA自身免疫性疾病患者36例(疾病对照组)和健康查体者30例(对照组)作为研究对象.根据临床资料计算DAS28评分,将RA组分为活动组和稳定组.采用酶联免疫吸附法(ELISA)检测血清中的抗MCV抗体、抗环瓜氨酸肽抗体(抗CCP抗体)及白细胞介素-6(IL-6)水平,采用免疫比浊法检测血清中C-反应蛋白(CRP)的浓度水平,并进行比较.结果 RA组抗MCV抗体的浓度水平显著高于非RA组和对照组(P均＜0.01);活动组抗MCV抗体的浓度水平显著高于稳定组和对照组(P均＜0.01);经Spearmau相关性分析,抗MCV抗体的浓度水平与CRP及IL-6呈正相关,而与DAS28无相关性;抗MCV抗体与抗CCP抗体的诊断效率无显著差异,但是灵敏度高于抗CCP抗体.结论 抗MCV抗体具有较高的灵敏度和特异性,并且能提示RA病情的活动情况,可以成为新的辅助RA诊断的血清学指标.     

靶向IL-6慢病毒介导RNA干扰实验性基因治疗类风湿关节炎

目的:探讨靶向小鼠IL-6基因RNA干扰慢病毒载体颗粒(LV-IL-6-RNAi)对小鼠巨噬细胞株J774A.1细胞IL-6表达的影响,及其对胶原诱导性关节炎(CIA)的治疗作用。方法:设计合成3对特异性针对小鼠IL-6基因的小干扰RNA(siRNA)序列,筛选最高效siRNA序,构建高效LV-IL-6-RNAi。LV-IL-6-RNAi感染小鼠巨噬细胞株J774A.1细胞,并设慢病毒阴性对照(LV-NC-RNAi)、培养基空白对照,RT-qPCR检测J774A.1细胞的IL-6、肿瘤坏死因子-α(TNF-α)和IL-1β mRNA相对表达水平,以检测LV-IL-6-RNAi体外干扰效率。取DBA/小鼠构建CIA模型并随机分为4组:LV-IL-6-RNAi组、LV-NC-RNAi组、甲氨蝶呤(MTX)阳性对照组、PBS空白对照组,分别关节腔注射LV-IL-6-RNAi、LV-NC-RNAi、腹腔注射MTX、PBS溶液,记录CIA小鼠关节炎评分,ELISA检测小鼠IL-6、IL-1β和TNF-α血清水平,组织病理检测炎症关节的炎症细胞浸润数。结果:成功构建了高效LV-IL-6-RNAi。...     

雷公藤多甙对胶原诱导性关节炎大鼠IL-10表达的影响

目的研究雷公藤多甙对类风湿关节炎大鼠的作用,探讨类风湿关节炎的发病机制及雷公藤多甙的作用机制,为临床应用提供理论基础。方法本实验应用鸡Ⅱ型胶原蛋白,建立胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠模型。应用酶联免疫吸附试验方法(ELISA)检测观察正常对照组、CIA模型组和雷公藤多甙治疗组大鼠血液及关节液中白细胞介素-10(interleukin-10,IL-10)的浓度。结果 ELISA检测结果显示,类风湿关节炎大鼠血清和关节液中IL-10含量明显减低,而雷公藤多甙可以升高IL-10的浓度,使之趋向正常。结论雷公藤多甙可有效减轻佐剂型关节炎大鼠的关节病变,并能使CIA大鼠血清、关节浸液IL-10分子水平增高。     

活动期类风湿关节炎患者血清及关节液白细胞介素-17水平变化及与有关实验室指标的关系

目的:分析类风湿关节炎(RA)患者外周血和关节液中T辅助细胞17(Thl7)相关细胞因子白介素-17(IL-17)水平,探讨其在RA中的变化特点及与实验室指标的相关性.方法:用ELISA检测50例活动期RA患者血清和其中15例关节液以及30例正常人血清IL-17水平.同时测定RA患者血清超敏C反应蛋白(hs-CRP)、...     

白细胞介素6及其受体对动脉粥样硬化的作用

动脉粥样硬化(AS)是一种多因素所致慢性血管炎症性疾病,炎症是AS的常见病理生理基础,在AS的发生、发展中起着重要作用。白细胞介素6(IL-6)是急性炎症反应因子和淋巴细胞刺激因子之一,其与受体(IL-6R)均参与AS的发生发展过程,可间接促炎促AS,又可直接促进AS,还能拮抗AS等。研究IL-6及其受体在AS中的分子机制,对AS的发生发展、靶向治疗和新药开发等方面的研究均有重要意义。     

The correlation between increased serum concentrations of interleukin-6 family cytokines and disease activity in rheumatoid arthritis patients

Purpose

This study was performed to determine whether the serum concentrations of interleukin (IL)-6 family cytokines are elevated in patients with rheumatoid arthritis (RA) and to investigate the relationship between IL-6 family cytokine levels and disease activity in RA patients.

Materials and Methods

We obtained serum samples from 40 patients with RA and 40 age- and sex-matched healthy controls, and we assessed the clinical parameters of disease activity, including the 28-joint disease activity score (DAS28) and C-reactive protein (CRP) levels. Serum samples from five patients with high disease activity (DAS28 > 5.1) were also collected at the eighth week of treatment. Serum concentrations of IL-6, IL-11, and leukemia inhibitory factor (LIF) were measured using an enzyme-linked immunosorbent assay (ELISA).

Results

Serum concentrations of IL-6 family cytokines, including IL-6, IL-11, and LIF, were significantly elevated in patients with RA compared to those of healthy controls. Although there was no significant relationship between IL-6 family cytokine levels and DAS28, the IL-6 levels of patients with RA showed a significant correlation with CRP levels. After eight weeks of medical treatment in patients with high disease activity, a decrease in DAS28 was associated with a significant decrease in the serum concentrations of IL-6 and IL-11.

Conclusion

The serum concentrations of IL-6 family cytokines were significantly elevated in patients with RA, and they decreased with medical treatment. These findings suggest a possible role for IL-6 family cytokines in the pathogenesis of RA.     

The role and therapeutic targeting of IL-6 in rheumatoid arthritis

Introduction: Rheumatoid arthritis (RA) is an autoimmune chronic disease with joint and systemic inflammation and it has been found that interleukin-6 (IL-6) plays a key role in RA. Indeed, various clinical studies have proved that the first-in-class IL-6 inhibitor, tocilizumab, a humanized anti-IL-6 receptor monoclonal antibody, showed outstanding efficacy in RA.

Areas covered: We review here the role of IL-6 in the inflammatory conditions and how IL-6 contributes to pathogenesis of RA, what induces IL-6 and how IL-6 expression is regulated. Furthermore, clinical studies of tocilizumab for RA are summarized,

Expert commentary: We review and discuss the prospects for future applications of IL-6 targeting therapy and new therapeutic strategies targeting IL-6. Finally, we discuss relevant issues with regard to the clinical management of IL-6 blockade in RA.     

Cytokine signal regulation and autoimmune disorders

Understanding of biological activities of cytokines and exquisite mechanism to regulate their functions has facilitated the therapeutic concept to restore the disequilibrium between pro-inflammatory cytokines and anti-inflammatory cytokines or cytokine inhibitors in some autoimmune inflammatory diseases such as rheumatoid arthritis (RA) and Crohn's disease. The application of molecular biology techniques to design monoclonal antibodies, soluble receptors, or receptor antagonists as therapeutic biologic agents made it possible to regulate the cytokine signals for the treatment of the diseases refractory to conventional therapies.

Japanese researchers have contributed considerably to the establishment of cytokine signal regulation in autoimmune diseases. In this article, Japanese studies of cytokine signal regulation, particularly for Interleukin-6 (IL-6) in autoimmune diseases are reviewed.     

Tocilizumab,A Humanized Anti-IL-6R Antibody,as an Emerging Therapeutic Option for Rheumatoid Arthritis: Molecular and Cellular Mechanistic Insights

Pro-inflammatory cytokines play a major role in the initiation and maintenance of joint inflammation and destruction in rheumatoid arthritis (RA). The therapeutic success of biologics targeting tumour necrosis factor-alpha (TNF-α), interleukin-1 (IL-1) and interleukin (IL)-6 receptor (IL-6R) has broadened the treatment options for RA. These agents have potential overlapping and discriminating biologic effects, as well as different pharmacological features. Tocilizumab (TCZ) is a humanized monoclonal antibody that binds and neutralizes IL-6R, resulting in the inhibition of various IL-6-mediated biological activities, including inflammation-related, immunomodulatory and tissue/matrix remodelling effects. Randomized, double-blind, controlled phase III studies and a number of early clinical observational studies have shown that treatment with TCZ results in rapid and sustained improvement in the signs and symptoms of RA among different patient populations. These studies have established the efficacy and safety of TCZ. Here, we review the pleiotropic functions of IL-6 and how it impinges on many aspects of RA pathogenesis, and highlight the clinical experience to date with TCZ as an emerging new treatment option for RA.     

Pathologic finding of increased expression of interleukin-17 in the synovial tissue of rheumatoid arthritis patients

Rheumatoid arthritis (RA) is a common autoimmune disease of chronic systemic inflammatory disorder that will affect multiple tissues and organs such as skin, heart or lungs; but it principally attacks the joints, producing a nonsuppurative inflammatory and proliferative synovitis that often progresses to major damaging of articular cartilage and joint ankylosis. Although the definite etiology is still unknown, recent studies suggest that T-helper cells (Th17) may play a pivotal role in the pathogenesis of RA. And interleukin-17 (IL-17), which is a cytokine of Th17 cells, may be a key factor in the occurrence of RA. The binding of IL-17 to specific receptor results in the expression of fibroblasts, endothelial and epithelial cells and also synthesis of several major factors such as tumor necrosis factor alpha (TNF-α), IL-1β that result in the structural damage of RA joints. Though some previous studies have shown that IL-17 exists in the synovium of RA, few has definite proof quantitatively by pathology about its existence in synovial membrane. This study comprised of 30 RA patients and 10 healthy control, pathologic study of the synovial membrane showed increased expression of IL-17 in the synovial tissue of RA patients, the intensity is compatible with clinical severity of disease as validated by DAS28 score and disease duration. Northern blot study also confirmed the increased expression of IL-17 in the synovial tissues. This study sheds further light that IL-17 may be a key factor in the pathogenesis of RA and a determinant of disease severity.     

地高辛标记人IL-2cDNA探针的制备及临床初步应用

目的：制备ＩＬ－２地高辛标记探针,探讨类风湿关节炎（ｒｈｅｕｍａｔｏｉｄａｒｔｈｒｉｔｉｓ,ＲＡ）患者外周血淋巴细胞ＩＬ－２ｍＲＮＡ的表达情况。方法 采用斑点杂交技术,对６例正常对照和１２例ＲＡ患者进行研究,结果：（１）从ｐＵＣ１２质粒子扩增制备了长度为１ｋｂ和ＩＬ－２ｃＤＮＡ片段,用地高辛标记后制备探针,其敏感性达１ｐｇ同源ＤＮＡ,且特异性较高,（２）１２例ＲＡ患者外周血淋巴细胞ＩＬ－２ｍＲＮＡ     

TRPC6在IL-1β诱导类风湿关节炎成纤维细胞样滑膜细胞增殖中的作用

目的:应用RNA干扰技术研究瞬时受体电位通道6(TRPC6)对IL-1β诱导的类风湿关节炎(RA)成纤维细胞样滑膜细胞(RA-FLS)增殖的影响。方法:RT-qPCR法检测RA和骨关节炎(OA)患者滑膜组织中TRPC6 mRNA的表达水平。组织块联合酶消化法培养RA-FLS。流式细胞术鉴定RA-FLS。将不同浓度(0、0.25、0.5、1、2、4、8、16μg/L)的重组人IL-1β与RA-FLS共培养36 h,CCK-8法检测细胞活力的改变;16μg/L的IL-1β作用RA-FLS不同时间(12、24、36、48、60、72 h),CCK-8法检测细胞活力的改变。特异性TRPC6-siRNA转染RA-FLS后,采用RT-qPCR和Western blotting检测沉默效率。在IL-1β存在和不存在的条件下,CCK-8法、Ed U标记法和流式细胞术检测TRPC6干扰组与对照组的细胞活力、Ed U阳性细胞比率和(G_2/M+S)期比率的差异。结果:RA患者滑膜组织中TRPC6的mRNA表达水平相对于OA患者明显增加(P0.05)。TRPC6-siRNA能显著降低RA-FLS中TRPC6 mRNA和蛋白的表达(P0.05)。IL-1β能诱导RA-FLS增殖(P0.05)。沉默TRPC6后,在IL-1β的诱导环境下,特异性干扰组RA-FLS的活力、Ed U阳性细胞比率和(G_2/M+S)期比率与空白组和对照组相比均明显降低(P0.05),而在不含IL-1β的条件下,干扰组与空白组和对照组相比差异均无统计学显著性。结论:TRPC6参与IL-1β诱导的RA-FLS增殖过程,沉默TRPC6能降低IL-1β诱导的RA-FLS增殖水平。     

A combination of interleukin-6 and its soluble receptor impairs sperm motility: implications in infertility associated with endometriosis

BACKGROUND: We previously reported that the level of interleukin (IL)-6 is increased in the peritoneal fluid of women with endometriosis. This study was undertaken to assess the effects of IL-6 and soluble IL-6 receptor (sIL-6R) on in vitro sperm motility. METHODS: Sperm (n = 20) were cultured with IL-6 or sIL-6R, or with a combination of both. After 24 h cultures, sperm motility was evaluated using a computer-assisted semen analysis system. Gene and protein expressions of IL-6, IL-6 receptor (IL-6R), and glycoprotein 130 (gp130) were examined in sperm by RT-PCR analysis and western blot analysis. RESULTS: Addition of IL-6 or sIL-6R individually to the culture media had no affect on sperm motion. However, adding a combination of IL-6 and sIL-6R dose-dependently reduced the percentage of motile and rapidly moving sperm. Adding anti-IL-6R antibody abolished these adverse effects. Sperm expressed the gp130 gene and protein, but not IL-6 or IL-6R. CONCLUSIONS: A combination of IL-6 and sIL-6R may be associated with gp130 expressed in the sperm and reduce sperm motility. IL-6 and sIL-6R may contribute to the pathogenesis of endometriosis-associated infertility.