白细胞介素-6受体在大鼠小脑间位核中的表达

我们以往的研究表明小脑皮层颗粒细胞表达白细胞介素-6受体(interleukin-6 receptor,IL-6R),该受体介导了白细胞介素-6(interleukin-6,IL-6)的神经保护作用。然而,大鼠小脑核团是否也表达IL-6R目前尚不清楚。为此,本研究应用RT-PCR、Western blot和免疫组织化学方法分别在基因、蛋白和细胞水平检测IL-6R在大鼠小脑间位核的表达。结果显示:(1)在小脑间位核PCR扩增产物的凝胶电泳图上,在365 bp位置可见一清晰的条带,与IL-6R mRNA预计扩增片断的长度一致;(2)在West-ern blot实验体系中,IL-6R免疫反应阳性条带出现在分子量为80 kD处;(3)免疫组织化学结果显示,小脑间位核中出现大量IL-6R免疫反应阳性细胞。以上结果表明:小脑间位核细胞在mRNA和蛋白水平均表达IL-6R,提示IL-6在小脑间位核内可能具有重要作用。     

白细胞介素18和白细胞介素10参与多囊卵巢综合征发病的初步研究

目的探讨血清炎症因子白细胞介素18（IL-18）、白细胞介素10（IL-10）与多囊卵巢综合征（PCOS）发病的关系。方法选取48例PCOS患者和32例对照,分为肥胖组与非肥胖组。测定血清空腹血糖（FBG）、空腹胰岛素（FINS）、卵泡刺激素（FSH）、黄体生成素（LH）和睾酮（T）;酶联免疫吸附试验（ELISA）方法检测血清IL-18、IL-10。结果 PCOS组血清IL-18水平高于对照组,IL-10水平低于对照组,差异有显著性;PCOS肥胖组与非肥胖组相比,IL-18水平升高,IL-10水平下降,差异有显著性;两PCOS组与各自对照组相比,IL-18水平升高,IL-10水平下降,差异有显著性。PCOS组患者血清IL-18与BMI、WHR、FINS及HOMA-IR呈显著正相关,血清IL-10与WHR、FINS及HOMA-IR呈显著负相关。结论 PCOS患者血清中IL-18水平明显升高,IL-10水平明显下降,并且在肥胖的PCOS患者中升高和下降的更明显。慢性炎症可能参与PCOS的发病,并且与胰岛素抵抗和肥胖有关。     

白细胞介素-15及其受体

白细胞介素-15(IL-15)是一种新发现的细胞因子,其生物学性质与IL-2相似。IL-15的受体由IL-15Rα、IL-2Rβ、IL-2Rγ组成,其中IL-2Rγ是参与IL-15信号传递过程的必要成分。IL-15和IL-2的体内表达及受体分布存在差异预示着IL-15在体内具有独特的生物学作用。     

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。     

白细胞介素-6及其受体与糖尿病肾病

白细胞介素－6（IL－6）是由多种细胞分泌的具有广泛生物活性的细胞因子，它通过与其肥体结合发挥作用。本文概述了IL－6及其受体的基因结构，来源及生物学作用，着重综述了糖尿病肾病（DN）时，活性增高的IL－6及其受体在促进DN发生，发展方面的意义，并检测肾局部IL－6mRNA表达上调，血尿IL－6及其可溶性受体活性增高与DN早期诊断，病情转归的相关性。     

白细胞介素-6在反复自然流产患者绒毛和蜕膜组织中的表达

目的:研究白细胞介素-6(IL-6)在反复自然流产患者绒毛与蜕膜组织中的表达情况,为防治反复自然流产提供理论依据.方法:应用免疫组织化学显色法和图像分析技术,对IL-6蛋白在反复自然流产患者(病例组)和正常人工流产者(对照组)绒毛和蜕膜组织中的表达进行定位和半定量分析;H-E染色法观察反复自然流产患者绒毛和蜕膜组织的形态学变化.结果:免疫组织化学显色显示,在绒毛组织中,IL-6蛋白定位于滋养层细胞的细胞质内,病例组的IL-6蛋白表达明显低于对照组;在蜕膜组织中,IL-6定位于蜕膜细胞和腺上皮细胞的细胞质内,且病例组蜕膜细胞的表达明显高于对照组,而2组在腺上皮细胞的表达无差异;H-E染色结果显示,病例组绒毛组织的滋养层变薄,细胞变性、坏死,细胞嗜酸性增强,绒毛中轴纤维化程度增强,蜕膜细胞失去细胞间连接,部分蜕膜细胞解体、核消失,细胞嗜酸性增强.结论:绒毛和蜕膜组织中IL-6蛋白表达的变化与反复自然流产有关,IL-6可能参与反复自然流产的病理过程.     

白细胞介素6受体

白细胞介素6受体(IL-6R)是造血生长因子受体家族的一员。它由一个80kD的配基结合链(IL-6R)和作为信号转导子的非配基结合链gp130组成。IL-6R既能以膜受体形式存在,也能以可溶性受体形式存在。可溶性白介素6受体(sIL-6R)同样具有膜受体的功能。白介素6受体的表达及调节在某些白介素6相关疾病的发病过程中起着重要作用。     

白细胞介素-5及其受体的分子生物学进展

由于IL-5受体拮抗剂对哮喘等慢性过敏性疾病具有潜在的应用前景,近年对IL-5及其受体的分子生物学进行了细致深入的研究。本文就IL-5及其受体的基因结构和分子特性、二者作用的分子机制及其信号转导方面的研究作一综述。     

白细胞介素13抑制肾小球系膜细胞增殖及白细胞介素6表达

目的:探讨白细胞介素13(IL-13)对体外培养的大鼠系膜细胞的增殖及其产生白细胞介素6(IL-6)的影响。方法:用四甲基偶氮唑(MTT)法测定系膜细胞增殖,用逆转录聚合酶链反应(RT-PCR)及酶联免疫吸附法(ELISA)测定系膜细胞IL-6mRNA表达及其蛋白水平。结果:IL-13在1、10、100μg/L浓度范围呈剂量依赖性地抑制系膜细胞的增殖;5%FCSRPMI1640培养条件下系膜细胞IL-6mRNA表达及IL-6分泌水平较低,脂多糖(LPS)可刺激系膜细胞IL-6mRNA的表达及提高IL-6分泌水平,而IL-13可抑制LPS诱导的系膜细胞IL-6分泌及其mRNA表达。结论:IL-13抑制体外培养的系膜细胞增殖及LPS诱导的系膜细胞IL-6的产生,IL-13可能对于肾小球肾炎的系膜细胞炎症反应具有拮抗作用。     

Endocytosis of interleukin-6—soluble interleukin-6 receptor complex and its intralysosomal degradation

Binding and internalization of interleukin-6—soluble interleukin-6 receptor complex by MDCK and MDCK-gp130 (transfected with gp 130 signal transductor) cells are studied. Binding of labeled complex depends on the concentration of interleukin-6; an effective internalization of the complex is shown. Binding and endocytosis of the complex are demonstrated in human hepatoma cells expressing interleukin-6 receptor and gp 130. These processes depend on the concentration of interleukin-6. The inhibitors of lysosomal functions ammonium chloride, monensin, and leupeptin suppress intralysosomal degradation of the complex, which confirms the important role of intralysosomal cleavage of the complex. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 11, pp. 527–529, November, 1997     

The clinical role of interleukin-6 and interleukin-6 soluble receptor in human follicular fluids

In order to investigate the role of interleukin-6 (IL-6) and interleukin-6 soluble receptor (sR) in human ovulation, we evaluated the concentrations in human follicular fluid and analyzed the correlation of IL-6 and IL-6 sR with oocyte maturation. The oocytes were obtained from the follicular fluid of 45 women undergoing in vitro fertilization and embryo transfer. The concentrations of IL-6 and IL-6 sR in follicular fluid were measured by ELISA. In addition, granulosa cells obtained from the follicular fluid were cultured and treated with forskolin and 12-o-tetradecanoylphorbol 13-acetate for 24–48 h. The concentration of IL-6 was significantly higher in the follicular fluid than in the serum (P<0.01). In contrast, the concentration of IL-6 sR was significantly lower in the follicular fluid than in the serum (P<0.001). The concentrations of IL-6 and IL-6 sR were significantly higher in the follicular fluid containing mature oocytes than in fluid containing immature oocytes (P<0.05). The production of IL-6 was markedly increased over the basal level after 24 h of treatment with forskolin(P<0.001) and 48 h of treatment (P<0.01) with cultured granulosa cells. Our data suggest that IL-6 and IL-6 sR may play an important role in follicular growth and development in human preovulatory processes. It is possible that IL-6 in particular may be regulated by cAMP. IL-6 and IL-6 sR might also be valuable biochemical markers in the evaluation of oocyte maturation. Received: 6 July 2002 / Accepted: 18 December 2002 Correspondence to Y. Kawano     

Astrocytic alterations in interleukin-6/Soluble interleukin-6 receptor alpha double-transgenic mice

Interleukin-6 (IL-6), a major cytokine with diverse effects on cells mainly of the immune and hematopoietic systems, has been linked to several neurological disorders such as acquired immune deficiency syndrome dementia, multiple sclerosis, and Alzheimer's disease. Central nervous system (CNS)-specific expression of IL-6 caused neurodegeneration, massive gliosis, and vascular proliferation in transgenic mice. However, the effects of systemically circulating IL-6 and its receptor IL-6Ralpha on the CNS are unknown. IL-6Ralpha is the specific component of the IL-6 receptor system and hence an important co-factor of IL-6. IL-6Ralpha is bioactive in a membrane-bound and in a soluble (s) form. We investigated the effects of systemically elevated levels of either human IL-6 or human sIL-6Ralpha or both on the CNS of transgenic mice. Although IL-6 and sIL-6Ralpha single transgenic mice were free of neurological disease, IL-6/sIL-6Ralpha double-transgenic mice showed neurological signs, such as tremor, gait abnormalities, and paresis. However, these mice also frequently showed prominent general weakness probably because of the systemic effects of IL-6/IL-6Ralpha such as liver damage and plasmacytomas. IL-6/sIL-6Ralpha transgenic mice exhibited massive reactive gliosis. Lack of signs of neuronal breakdown versus ample astrogliosis suggested that astrocytes were selectively affected in these mice. There was neither vascular proliferation nor inflammatory infiltration. Ultrastructural analysis revealed blood-brain barrier (BBB) changes manifested by hydropic astrocytic end-feet. However, albumin immunohistochemistry did not reveal major BBB leakage. Our results indicate that increased and constitutive systemic expression of IL-6 together with its soluble receptor sIL-6Ralpha is less harmful to the brain than to other organs. The BBB remains primarily intact. IL-6/IL-6Ralpha, however, might be directly responsible for the selective activation of astrocytes.     

Involvement of gp130/interleukin-6 receptor transducing component in interleukin-11 receptor

The recently cloned interleukin (IL)-11 displays many biological properties in common with those reported for IL-6. In order to analyze the nature and the functionality of the IL-11 receptor we developed a proliferative assay using the human multifactor-dependent cell line TF1. We showed that a blocking monoclonal antibody GPX7 raised against the gp130/IL-6 receptor transducing subunit was also able to inhibit the IL-11-triggered TF1 line proliferation. In addition, involvement of gp130 in IL-11 signaling was demonstrated by an induction of the transducing protein phosphorylation in response to IL-11, as observed for IL-6. In contrast, the blocking monoclonal antibody B-R6, which recognized the gp80/IL-6 binding subunit, failed to interfere with the IL-11 proliferative signal in the TF1 cell line. Similarly, we did not observe any competition between IL-6 and IL-11 for a putative common binding site on the cell surface. These results suggest that the IL-11 binding component is different from the gp80/IL-6 receptor. In conclusion, IL-11, along with IL-6, leukemia inhibitory factor, oncostatin M and ciliary neurotrophic factor, belongs to the same family of cytokines, using gp130 as a transducing protein.     

Immunohistochemical localization of interleukin-6 and its receptor in benign, premalignant and malignant prostate tissue

Interleukin-6 (IL-6) is a pleiotropic cytokine that interacts with its receptor in prostate cells, thus regulating proliferative response and differentiation. It also activates the human androgen receptor in prostate cancer cell lines. In order to assess the significance of these findings in vivo, the expression of key elements of the IL-6 signalling pathway, IL-6 and its receptor, was investigated in tissue samples obtained on radical prostatectomy from prostate cancer patients. IL-6 immunohistochemistry was performed on 17 frozen prostate cancer specimens. IL-6 receptor immunostaining was evaluated in 21 paraffin-embedded prostate tumour specimens. In both groups, adjacent areas of high-grade prostatic intraepithelial neoplasia and benign tissue were also investigated. In benign prostatic epithelium, IL-6 was localized predominantly in basal cells, whereas in prostate cancer tissues more IL-6-positive glandular cells were identified. No IL-6 expression was detected in stromal cells on immunohistochemistry, although IL-6 protein was measured in the supernatants obtained from cultured stromal cells by ELISA. IL-6 receptor was expressed in benign prostatic tissue in both epithelial and stromal cells. Furthermore, IL-6 receptor expression was observed in all tumour specimens investigated and the majority of Gleason patterns analysed had more than 50% of cells showing a positive reaction. IL-6 and IL-6 receptor expression patterns in high-grade prostatic intraepithelial neoplasia lesions were similar to those observed in tumour tissues. Taken together, the results of the present study imply that there are paracrine and autocrine IL-6 loops in benign and neoplastic prostate.     

成纤维细胞介导的人白细胞介素6基因疗法的建立及其体内白细胞介素6分泌水平的实验研究

细胞因子基因治疗是近年来生物治疗的重要进展。本文以人IL-6基因为治疗目的基因,以成纤维细胞为载体细胞,建立了人IL-6基因疗法的实验模型,并动态观察了其体内IL-6分泌水平.将650bp的人IL-6 cDNA 插入到携有Neo~R 基因的表达载体BCMGNeo 的Xho I 位点上,对此重组表达载体进行限制性酶切鉴定。用磷酸钙共沉淀法将BCMGNeo-IL-6转入NIH3T3成纤维细胞中,通过G418抗性筛选、有限稀释和上清中IL-6活性的测定,从多株阳性克隆中筛选到一株高分泌IL-6(184.6U/ml)的克隆株。对此阳性克隆进行了Southern 杂交分析。将此阳性克隆体外扩增、包裹入胶原中,然后移植入小鼠腹腔内,可从小鼠血清中(直至移植后15天)检测出IL-6,明成纤维细胞能成功地将IL-6基因导入体内并有效表达,证明该基因疗法是可行的。     

Characterization of anti-mouse interleukin-6 receptor antibody

Hybridoma that produces rat anti-mouse interleukin 6 receptor (IL-6R) antibody, MR16-1, was established by the fusion of mouse P3U1 myeloma cells and spleen cells from mouse soluble IL-6R (sIL-6R)-immunized Wistar rat. In the present study, we examined the characteristics of MR16-1 in vitro and in vivo. MR16-1 bound to mouse sIL-6R dose-dependently. MR16-1 suppressed IL-6-induced proliferation of 7TD1 cells in a dose-dependent manner and this inhibitory effect was reversed by the addition of a higher concentration of IL-6. Cross-reactivity study using T cells from mouse, rat, and human revealed that MR16-1 did not cross-react with human and rat IL-6R. Binding region analysis using several human-mouse chimeric IL-6Rs showed that half of the fibronectin domain II of mouse IL-6R (amino acids 214-285) was required for MR16-1 binding. Furthermore, MR16-1 completely suppressed IL-6-induced antibody production in DNP-KLH immunized mice. These lines of evidence demonstrate that MR16-1 is useful to investigate the physiological and pathological roles of IL-6 and sIL-6R in mice.     

The effect of interleukin-6 and the interleukin-6 receptor on glucose transport in mouse skeletal muscle

Exercise results in an increase in interleukin-6 (IL-6), its receptor (IL-6R) and skeletal muscle glucose transport. Interleukin-6 has been found to have equivocal effects on glucose transport, with no studies, to our knowledge, investigating any potential role of IL-6R. In the present study, we hypothesized that a combined preparation of IL-6 and soluble IL-6R (sIL-6R) would stimulate glucose transport. Mouse soleus muscles were incubated with physiological and supraphysiological concentrations of IL-6 and a combination of IL-6 and sIL-6R. Total and phosphorylated AMP-activated protein kinase (AMPK) and Protein Kinase B (PKB/Akt) were also measured by Western blotting. Exposure to both physiological (80 pg ml⁻¹) and supraphysiological IL-6 (120 ng ml⁻¹) had no effect on glucose transport. At physiological levels, exposure to a combination of IL-6 and sIL-6R (32 ng ml⁻¹) resulted in a 1.4-fold increase ( P < 0.05) in basal glucose transport with no change to the phosphorylation of AMPK. Exposure to supraphysiological levels of IL-6 and sIL-6R (120 ng ml⁻¹) resulted in an approximately twofold increase ( P < 0.05) in basal glucose transport and an increase ( P < 0.05) in AMPK phosphorylation. No effect of IL-6 or sIL-6R was observed on insulin-stimulated glucose transport. These findings demonstrate that, while IL-6 alone does not stimulate glucose transport in mouse soleus muscle, when sIL-6R is introduced glucose transport is directly stimulated, partly through AMPK-dependent signalling.     

Modelling of fish interleukin-1 and its receptor

This paper presents original data regarding: the three-dimensional modelling of rainbow trout (Oncorhynchus mykiss) and sea bass (Dicentrarchus labrax) IL-1beta, the modelling of trout IL-1 receptor type I (IL-1RI) and the modelling of trout IL-1beta bound to its receptor. The 3D models of trout and sea bass IL-1beta molecules were predicted by comparison with those already available of human and mouse, and, in both cases, a structure consisting of a beta-trefoil fold was obtained, a motif well conserved during evolution of IL-1beta-related proteins. Moreover, a model for the rainbow trout IL-1 receptor alone and complexed with IL-1beta was predicted and compared to the murine model. Both ligand-receptor complex models were compared with the known crystal structure of the human IL-1beta/IL-1R complex. A cross-interaction of trout IL-1beta with the mouse receptor was also simulated. Such analysis and the predicted interaction energies calculated from the models have helped to explain the different biological efficacies of mammalian and fish IL-1beta molecules in assays based on mammalian target cells. The obtained data are discussed on the basis of evolutionary and applied perspectives.