姜黄素对脂多糖诱导人支气管上皮细胞 丝裂素活化蛋白激酶信号通路的影响

目的 探讨姜黄素对脂多糖(LPS)诱导人支气管上皮细胞三磷酸肌醇激酶(PI3K)、蛋白激酶B (Akt)、外调节蛋白激酶(ERK)、核转录因子-κB(NF-κB)、不规则趋化因子(CX3CL1)表达的影响.方法 体外培养人支气管上皮细胞24 h后,用计算机生成的随机分配法表分为空白组(CK)、LPS(10 mg/L)组、PD组 (LPS+ ERK抑制剂PD98059,25μmol/L)、LY组(LPS+ PI3K/Akt抑制剂LY294002,20μmol/L)、PDTC组 (LPS+ NF-κB抑制剂PDTC,100μmol/L)、cur组(LPS+姜黄素,10μmol/L)6组.各组阻断剂预处理30 min后给予LPS刺激,加药后作用4 h,用蛋白质免疫印迹试验(Western Blot)检测各组PI3K、Akt、ERK、NF-κB、CX3CL1表达,测定各条带吸光度(A)值.结果 与CK组比较,LPS组PI3K、Akt、ERK、NF-κB、CX3CL1表达均显著升高〔PI3K(A值):0.68±0.05比0.49±0.09,Akt(A值):0.54±0.03比0.30±0.06,ERK(A值):1.09±0.09比0.74±0.05、NF-κB(A值):1.87±0.15比0.57±0.30,CX3CL1(A值):0.62±0.12比0.34±0.00,均P<0.05〕.与LPS组比较,PD组、PDTC组PI3K、Akt、ERK、NF-κB、CX3CL1表达均明显下降〔PI3K(A值):0.37±0.06、0.38±0.16比0.68±0.05,Akt(A值):0.33±0.07、0.33±0.12比0.54±0.03,ERK(A值):0.67±0.05、0.82±0.26比1.09±0.09,NF-κB(A值):0.73±0.19、0.97±0.41比1.87±0.15,CX3CL1(A值):0.43±0.07、0.32±0.03比0.62±0.12,均P<0.05〕;cur组能明显抑制PI3K、Akt、ERK、NF-κB表达〔PI3K(A值):0.44±0.04比0.68±0.05, Akt(A 值):0.30±0.10 比 0.54±0.03,ERK(A 值):0.78±0.05 比 1.09±0.09,NF-κB(A 值):0.78±0.17 比1.87±0.15,均P<0.05〕.结论 人支气管上皮细胞存在LPS—ERK、NF-κB—CX3CL1信号通路,姜黄素可抑制LPS诱导人支气管上皮细胞后PI3K/Akt、ERK、NF-κB的表达.     

类风湿关节炎患者血浆可溶型不规则趋化因子检测的临床意义

目的通过检测血浆中的可溶型不规则趋化因子（sFKN）的水平,探讨其与类风湿关节炎（RA）发病的关系及其与疾病活动的相关性。方法应用ESLIA方法分别测定30例RA活动组、30例RA缓解组及30名健康对照组的血浆sFKN水平,并同时应用ESLIA方法测定类风湿因子（RF）、抗环瓜氨酸肽（CCP）抗体及记录病程、晨僵时间、关节压痛数、关节肿胀数、Ritchter指数等活动性指标。结果 RA组血浆sFKN水平明显高于健康对照组,且RA活动组血浆sFKN明显高于RA缓解组（均P﹤0.05）。血浆sFKN水平与RF、抗CCP抗体及ESR、C反应蛋白（CRP）、关节肿胀数、关节压痛数、Ritcher指数等活动性指标成显著正相关性（均P〈0.01）,与晨僵时间、病程无相关性（均P〉0.05）。结论血浆sFKN可能参与了RA的病理生理过程且有助于RA的诊断,并可作为判断RA活动性的重要指标之一。     

Hypertrophy of the ligament flavum in degenerative lumbar stenosis associated with the increased expression of fractalkine (CX3CL1)/CX3CR1 chemokine

Abstract

Fractalkine (CX3CL1) and its receptor (CX3CR1) comprise a chemokine system involved in leukocyte recruitment and adhesion in chronic inflammatory disease, but its role in spinal diseases is unknown. The purpose of this study is to investigate the role of CX3CL1/CX3CR1 chemokine on hypertrophy of the ligamentum flavum (LF) in degenerative lumbar stenosis (DLS) compared with that of non-degenerative spinal condition (NDS) of the lumbar spine and correlation between expression of CX3CL1/CX3CR1 chemokine and thickness of LF. The mRNA concentrations of CX3CL1/CX3CR1 chemokine were analyzed in the surgically obtained LF specimens from DLS (n?=?10) and NDS (n?=?11) by real-time PCR. The localization of CX3CL1/CX3CR1 chemokine within the LF was determined using immunohistochemical study. Plasma levels of soluble FKN (sFKN) were measured by enzyme-linked immunosorbent assay, respectively. The thickness of the LF was measured with axial T1-weighted MRI. The cells that express CX3CL1/CX3CR1 chemokine ratio in the LF observed in DLS group were substantially higher than in NDS group. In ELISA, the plasma levels of sFKN was significantly increased in DLS compared with patients in the other groups (p?=?0.006). There was greater CX3CL1/CX3CR1 expression in DLS as quantified by RT-PCR (p?=?0.004, 0.010). Thickness of LF in patients was significantly correlated with serum CX3CL1 level (R^2?=?0.824, p?=?0.003) and with mRNA expression of CX3CL1/CX3CR1 (R^2?=?0.671, p?=?0.000) (R^2?=?0.514, p?=?0.001). This study identified for the first time that increases in CX3CL1 and CX3CR1-expressing cells are significantly related to LF hypertrophy.     

Aspirin inhibits tumor necrosis factor-α-stimulated fractalkine expression in human umbilical vein endothelial cells

Background Fractalkine is an important chemokine mediating local monocyte accumulation and inflammatory reactions in the vascular wall. Aspirin inhibits inflammatory cytokine expression closely related to atherosclerosis through the way independent of platelet and cyclooxygenase （COX）. There has been no report about the effect of aspirin on fractalkine expression. We aimed to determine the fractalkine expression in human umbilical vein endothelial cell （HUVEC） stimulated by tumor necrosis factor （TNF）-α and the effect of aspirin intervention.
Methods Six of 8 HUVEC groups received either different concentrations of aspirin （0.02, 0.2, 1.0, 5.0 mmol/L） or 40 μmol/L pyrrolidinecarbodithioc acid （PDTC） or 0.5 μmol/L NS-398. The other two groups were negative control and positive control （TNF-α-stimulated）. After being incubated for 24 hours, cells of the 8 groups except the negative control one were stimulated with TNF-α （4 ng/ml） for another 24 hours. After that, the cells were collected for RNA isolation and protein extraction.
Results Both mRNA and protein expressions of fractalkine in HUVEC were upregulated by 4 ng/ml TNF-α stimulation. Aspirin inhibited fractalkine expression in a dose-dependent manner at mRNA and protein levels. Nuclear factor-kappa B inhibitor, PDTC, effectively decreased the fractalkine expression. Fractalkine expression was not influenced by COX-2 selective inhibitor NS-398. COX-1 protein expression was not changed by either TNF-α stimulation or aspirin, PDTC, NS-398 intervention. Both mRNA and protein expression of COX-2 in HUVEC were upregulated by 4 ng/ml TNF-α stimulation. Aspirin decreased COX-2 expression in a dose-dependent manner at mRNA and protein levels.
Conclusions TNF-α-stimulated fractalkine expression is suppressed by aspirin in a dose-dependent manner through the nuclear factor-kappa B p65 pathway.     

Fractalkine/CX3CR1: why a single chemokine-receptor duo bears a major and unique therapeutic potential

Importance of the field: Fractalkine, also known as CX3CL1, is the unique member of the fourth class of chemokines and mediates both chemotaxis and adhesion of inflammatory cells via its highly selective receptor CX3CR1. Fractalkine mediates inflammatory responses and pain sensation and is involved in the pathogenesis and progression of numerous inflammatory disorders and malignancies.

Areas covered in this review: We performed a Medline/PubMed search to detect all published studies that explored the role of fractalkine and CX3CR1 and the possibilities of therapeutic intervention in the fractalkine/CX3CR1 axis in a wide range of clinical disorders, using CX3CR1 blocking antibodies, different fractalkine antagonists, CX3CR1 depletion or transfection of fractalkine expression vectors.

What the reader will gain: This review summarizes the role of fractalkine and its receptor CX3CR1 in various diseases, focusing on their high potential as novel therapeutic targets, with special emphasis on pancreatic diseases.

Take home message: The reviewed studies provide promising results demonstrating fractalkine and CX3CR1 as potential target molecules for future therapeutics that may attenuate pain, inflammation and furthermore serve as an anti-cancer therapy. However, to date, no therapeutics targeting fractalkine or CX3CR1 are in clinical use.     

Therapeutic potential of the chemokine–receptor duo fractalkine/CX3CR1: an update

Introduction: The chemokine fractalkine/CX3CL1 and its highly selective receptor CX3CR1 mediate critical physiological events during inflammatory responses. The fractalkine/CX3CR1 axis has been shown to play a key role in the pathogenesis and the progression of a large number of diseases in which imbalance of the immune response is frequently seen. Since our last review published in early 2010, the fractalkine/CX3CR1 axis has gained vast attention as a potential therapeutic target in the scientific community, which can be clearly seen in the large number of studies that have been published on this issue since then.

Areas covered: A Medline/PubMed search was performed to detect all recently published studies on the role of the fractalkine/CX3CR1 axis as a therapeutic target in a wide range of clinical diseases.

Expert opinion: Recently published studies further underline the high potential of the fractalkine/CX3CR1 axis as a major target for future treatment of pain, inflammation and cancer. However, no clinical trials on novel therapeutics targeting fractalkine or CX3CR1 have been initiated so far, so that the fractalkine/CX3CR1 axis does still not find application in daily clinical practice.     

The CX3C chemokine fractalkine in allergic asthma and rhinitis

BACKGROUND: Unlike other chemokines, fractalkine is expressed as a membrane-bound form, mainly on endothelial and epithelial cells, and can be shed as a soluble chemotactic form. Fractalkine can capture leukocytes expressing its receptor (CX(3)CR(1)), including T lymphocytes, rapidly and firmly in an integrin-independent manner. Because of its dual activity, fractalkine plays a major role in the transendothelial and transepithelial migration of leukocytes during inflammation. OBJECTIVE: We sought to study the fractalkine-CX(3)CR(1) axis in patients with allergic airways diseases. METHODS: Plasma fractalkine levels were measured by means of ELISA in 19 control subjects and 55 patients with symptomatic allergic rhinitis, asthma, or both, and CX(3)CR(1) function was studied by using triple-color flow cytometry in circulating T-lymphocyte subpopulations. Segmental allergen challenge was performed in 16 allergic asthmatic patients to analyze fractalkine expression and inflammatory cell recruitment in bronchoalveolar lavage fluid and bronchial biopsy specimens. RESULTS: Compared with control subjects, patients with symptomatic allergic rhinitis and asthmatic patients had increased circulating fractalkine levels, and CX(3)CR(1) function was upregulated in circulating CD4(+) T lymphocytes. Twenty-four hours after segmental allergen challenge, bronchoalveolar lavage fluid soluble fractalkine concentrations increased and correlated with the total number of recruited cells. Bronchial epithelial and endothelial cells expressed high levels of the membrane-bound form of fractalkine before and after challenge. CONCLUSION: Allergic asthma and rhinitis are associated with systemic and bronchial upregulation of the chemotactic axis fractalkine-CX(3)CR(1). This might contribute to the rapid recruitment of circulating CD4(+) T lymphocytes in the airways after allergen stimulation.     

Respiratory syncytial virus(RSV)-induced allergy may be controlled by IL-4 and CX3C fractalkine antagonists and CpG ODN as adjuvant: hypothesis and implications for treatment

Based on the hypothesis that respiratory syncytial virus (RSV) sG protein causes allergy in patients, it is suggested that treatment of RSV patients with antagonists of IL-4 and FKN early in infection will prevent the increased level of IL-4 in the serum. Together with CpG ODNs that induce Toll-like receptor 9⁺ (TLR9⁺) plasmacytoid dendritic cells to release type I IFN-α and -β will reactivate the inhibited Th1 cells and the antiviral cytotoxic T leukocytes. In addition, binding of CpG ODNs to TLR9⁺ B cells will stop IgE synthesis and antiviral IgG and IgA will continue. Together, the IL-4 and FKN antagonists and CpG ODNs reactivate the adaptive immune response to clear the virus and protect the patient from a second RSV infection. It is also suggested that the less-pathogenic RSV strain Long may be a candidate for vaccine development after deletion of the FKN and superantigen domains from the G gene.