血清FKN、APC与老年社区获得性肺炎患者病情和预后不良的关系

摘要 目的：探讨血清分形趋化因子（FKN）、活化蛋白C（APC）与老年社区获得性肺炎（CAP）患者病情和预后不良的关系。方法：选取2020年1月～2023年1月潍坊市人民医院收治的314例老年CAP患者为CAP组,根据病情程度分为低危组104例、中危组123例、高危组87例,根据入院30d生存状况分为死亡组65例和存活组249例,另选取同期100名体检健康老年人为对照组。采用酶联免疫吸附法检测血清FKN、APC水平。采用受试者工作特征（ROC）曲线分析血清FKN、APC水平对老年CAP患者预后不良的预测价值。通过多因素Logistic回归分析老年CAP患者预后不良的影响因素。结果：与对照组比较,CAP组血清FKN水平升高,APC水平降低（P＜0.05）。低危组、中危组、高危组老年CAP患者血清FKN水平依次升高,APC水平依次降低（P＜0.05）。多因素Logistic回归分析显示,病情高危、机械通气和C反应蛋白（CRP）、FKN升高为老年CAP患者预后不良的独立危险因素,APC升高为其独立保护因素（P＜0.05）。ROC曲线分析显示,FKN、APC水平单独和联合预测老年CAP患者预后不良的曲线下面积分别为0.783、0.789、0.870,二者联合对老年CAP患者预后不良的预测价值大于各指标单独预测。结论：血清FKN水平升高和APC水平降低参与着老年CAP患者病情进展,血清FKN联合APC能较好地预测老年CAP患者预后不良。     

血清Fractalkine、Apelin水平与糖尿病视网膜病变患者血糖、血脂以及病程的关系研究

目的:研究血清Fractalkine(FKN)、爱帕琳肽(Apelin)水平与糖尿病视网膜病变(DR)患者血糖、血脂以及病程的关系。方法:选取我院于2015年1月至2016年12月收治的160例糖尿病患者为研究对象,行眼底荧光造影、裂隙灯显微镜检查,按照检查结果将其区分为非增生型DR组(稳定组,43例)、背景期DR组(背景组,62例)和增殖期DR组(增殖组,55例),另外于同期选取我院40例健康体检者为健康对照组(健康组),测量4组血清FKN、Apelin、空腹血糖(FPG)、餐后2h血糖(2hPG)、糖化血红蛋白(HbA1c)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)和总胆固醇(TC)水平,使用Pearson相关性分析分析血清FKN、Apelin与FPG、2hPG、HbA1c、HDL-C、LDL-C、TG、TC、糖尿病病程的相关性。结果:血清FKN、Apelin水平比较:增殖组背景组稳定组健康组,各组间比较差异具有统计学意义(P0.05);血清FPG、2hPG、HbA1c、LDL-C、TG、TC水平比较:增殖组背景组稳定组健康组,各组间比较差异具有统计学意义(P0.05);血清HDL-C水平比较:健康组稳定组背景组增殖组,各组间比较差异具有统计学意义(P0.05);采用Pearson相关性分析显示,血清FKN水平与FPG、2hPG、HbA1c、LDL-C、TG、TC、糖尿病病程呈正相关性(r=0.321、0.215、0.645、0.154、0.215、0.325、0.578,P0.05),与HDL-C呈负相关性(r=-0.547,P0.05);血清Apelin水平与FPG、2hPG、HbA1c、LDL-C、TG、TC、糖尿病病程呈正相关性(r=0.245、0.574、0.951、0.357、0.357、0.159、0.546,P0.05),与HDL-C呈负相关性(r=-0.459,P0.05);糖尿病病程、HbA1c、LDL-C、HDL-C、FKN和Apelin为DR病程的相关影响因素。结论:糖尿病伴发DR患者血清FKN、Apelin水平随着病程的加重逐渐增加,且这两种因子的水平与患者血糖、血脂代谢关系密切。     

Use of cocultured cell systems to elucidate chemokine-dependent neuronal/microglial interactions: control of microglial activation

In order to understand processes involved in central nervous system inflammatory diseases, a critical appreciation of mechanisms involved in the control of immune function in the brain is needed. Microglial cells are watchful eyes for unusual events and detecting the presence of pathogens but are also alert to signals emanating from damaged neurons. Fractalkine (CX3CL1) is a chemokine which is expressed predominantly in the central nervous system, being localized on neurons, while its receptor, CX3CR1, is found on microglial cells. We have developed a strategy to investigate the role of this chemokine in neuronal-microglia interactions. Because fractalkine is expressed both as a soluble and as a membrane-attached protein, we have established various protocols involving different levels of cell-to-cell communication. Three experimental systems were instituted, including (1) a conditioned medium transfer system in which no cell-cell communication or contact is possible, (2) a transwell system that permits cell-contact-independent communication through diffusible soluble factors only, and (3) a coculture system allowing cell-to-cell communication via direct microglial-neuronal contacts. Using these in vitro cocultured systems, we have investigated the role of a soluble and/or cell-associated chemokine, such as fractalkine, in order to obtain insights into the role of glia-neuron interactions in cerebral inflammation.     

Decreased Fractalkine and Increased IP-10 Expression in Aged Brain of APP<Subscript>swe</Subscript> Transgenic Mice

Chemokines and their receptors have been strongly implicated in the inflammatory process and pathogenesis of the neurodegenerative disorders, such as Alzheimer’s disease (AD). In the present study, we examined the expression of chemokines, fractalkine, interferon-inducible protein-10 (IP-10) and macrophage inflammatory protein-1α (MIP-1α) by immunohistochemistry in the brain of transgenic mice APP_SWE (Tg2576) at ages of 9, 11, and 17 months, which over-express a mutated form of human amyloid precursor protein (APP). Decreased fractalkine and increased IP-10 expression in cerebral cortex and hippocampus were found at ages of 9 and 17 months in Tg2576 mice when compared with age-matched control mice. On the contrary, MIP-1α expression showed no difference between Tg2576 mice and aged controls and was not influenced by ages. β-amyloid (Aβ) positive plaques were co-located with the intense IP-10 expression. The finding suggests fractalkine and IP-10 may participate in the pathogenesis of AD; and could be new therapeutic strategies for neuroprotection.     

Autocrine role of vascular IL-15 in intimal thickening

Interleukin 15 (IL-15) is a pro-inflammatory cytokine that modulates T cell recruitment and activation, independent of antigen. It has been detected in human atherosclerotic plaques and atherosclerotic plaques of apoE-/- mice. IL-15 regulates fractalkine (FKN)-CX3CR1 chemokine signaling which is involved in atherogenesis and promotes SMC proliferation. We investigated the role of IL-15 in intimal thickening after arterial injury. Treatment of serum-stimulated SMC with IL-15 in vitro attenuated proliferation and suppressed CX3CR1 and FKN mRNA expression. The role of endogenous IL-15 in vivo was investigated in injured carotid arteries of mice. Periadventitial arterial injury resulted in increased IL-15 expression in the media and neointima, paralleled by increased IL-15 receptor alpha expression. Blockade of endogenous IL-15 increased intimal thickening. FKN and CX3CR1 expression increased after injury and were further augmented after IL-15 blockade. These data suggest that endogenous IL-15 attenuated intimal thickening after arterial injury. The potential mechanism of action is suppression of CX3CR1 signaling.     

Proinflammatory Secreted Phospholipase A2 Type IIA (sPLA-IIA) Induces Integrin Activation through Direct Binding to a Newly Identified Binding Site (Site 2) in Integrins αvβ3, α4β1, and α5β1

Integrins are activated by signaling from inside the cell (inside-out signaling) through global conformational changes of integrins. We recently discovered that fractalkine activates integrins in the absence of CX3CR1 through the direct binding of fractalkine to a ligand-binding site in the integrin headpiece (site 2) that is distinct from the classical RGD-binding site (site 1). We propose that fractalkine binding to the newly identified site 2 induces activation of site 1 though conformational changes (in an allosteric mechanism). We reasoned that site 2-mediated activation of integrins is not limited to fractalkine. Human secreted phospholipase A2 type IIA (sPLA2-IIA), a proinflammatory protein, binds to integrins αvβ3 and α4β1 (site 1), and this interaction initiates a signaling pathway that leads to cell proliferation and inflammation. Human sPLA2-IIA does not bind to M-type receptor very well. Here we describe that sPLA2-IIA directly activated purified soluble integrin αvβ3 and transmembrane αvβ3 on the cell surface. This activation did not require catalytic activity or M-type receptor. Docking simulation predicted that sPLA2-IIA binds to site 2 in the closed-headpiece of αvβ3. A peptide from site 2 of integrin β1 specifically bound to sPLA2-IIA and suppressed sPLA2-IIA-induced integrin activation. This suggests that sPLA2-IIA activates αvβ3 through binding to site 2. sPLA2-IIA also activated integrins α4β1 and α5β1 in a site 2-mediated manner. We recently identified small compounds that bind to sPLA2-IIA and suppress integrin-sPLA2-IIA interaction (e.g. compound 21 (Cmpd21)). Cmpd21 effectively suppressed sPLA2-IIA-induced integrin activation. These results define a novel mechanism of proinflammatory action of sPLA2-IIA through integrin activation.     

The anti-atherosclerotic effect of tanshinone IIA is associated with the inhibition of TNF-α-induced VCAM-1, ICAM-1 and CX3CL1 expression

Tanshinone IIA is one of the major diterpenes in Salvia miltiorrhiza. The inhibitory effect of Tanshinone IIA on atherosclerosis has been reported, but the underlying mechanism is not fully understood. The present study aimed to study the anti-atherosclerosis effect of Tanshinone IIA on the adhesion of monocytes to vascular endothelial cells and related mechanism. Results showed that Tanshinone IIA, at the concentrations without cytotoxic effect, dose-dependently inhibited the adhesion of THP-1 monocytes to the TNF-α-stimulated human vascular endothelial cells. The expressions of cell adhesion molecules including VCAM-1, ICAM-1 and E-selectin were induced by TNF-α in HUVECs at both the mRNA and protein levels. The mRNA and protein expressions of VCAM-1 and ICAM-1, but not E-selectin, were both significantly suppressed by Tanshinone IIA in a dose dependent manner. In addition, the TNF-α-induced mRNA expression of fractalkine/CX3CL1 and the level of soluble fractalkine were both reduced by Tanshinone IIA. We also found that Tanshinone IIA significantly inhibited TNF-α-induced nuclear translocation of NF-κB which was resulted from the inhibitory effect of Tanshinone IIA on the TNF-α-activated phosphorylation of IKKα, IKKβ, IκB and NF-κB. As one of the major components of Salvia miltiorrhiza, Tanshinone IIA alone exerted more potent effect on inhibiting the adhesion of monocytes to vascular endothelial cells when compared with Salvia miltiorrhiza. All together, these results demonstrate a novel underlying mechanism for the anti-inflammatory effect of Tanshinone IIA by modulating TNF-α-induced expression of VCAM-1, ICAM-1 and fractalkine through inhibition of TNF-α-induced activation of IKK/NF-κB signaling pathway in human vascular endothelial cells.     

弥漫性毒性甲状腺肿患者血清高半胱氨酸蛋白61及Fractalkine水平的表达及其临床意义

摘要 目的：探讨弥漫性毒性甲状腺肿（GD）患者血清高半胱氨酸蛋白61（CYR61）、Fractalkine水平的表达及其临床意义。方法：选取2018年3月～2021年10月河北省邯郸市中心医院收治的57例GD患者作为研究组。另取同期健康体检者50例。采集所有受试者的静脉血,检测血清CYR61、Fractalkine水平,甲状腺功能及甲状腺自身抗体相关指标。采用Pearson检验分析GD患者血清CYR61、Fractalkine水平与甲状腺功能及甲状腺自身抗体相关指标的相关性。结果：研究组血清CYR61、Fractalkine水平均高于对照组,差异均有统计学意义（均P＜0.05）。研究组血清游离三碘甲腺原氨酸（FT3）、游离四碘甲腺原氨酸（FT4）均高于对照组,而促甲状腺激素（TSH）水平低于对照组,差异均有统计学意义（均P＜0.05）。研究组抗甲状腺球蛋白抗体（TGAb）、甲状腺过氧化物酶抗体（TPOAb）及促甲状腺激素受体抗体（TRAb）水平均高于对照组,差异均有统计学意义（均P＜0.05）。经Pearson相关性分析发现,GD患者血清CYR61、Fractalkine水平与FT3、FT4、TGAb、TPOAb、TRAb水平均呈正相关,而与血清TSH水平呈负相关（均P＜0.05）。结论：GD患者血清CYR61、Fractalkine水平异常高表达,且与患者甲状腺功能及甲状腺自身抗体有关。     

Fractalkine attenuates excito-neurotoxicity via microglial clearance of damaged neurons and antioxidant enzyme heme oxygenase-1 expression

Glutamate-induced excito-neurotoxicity likely contributes to non-cell autonomous neuronal death in neurodegenerative diseases. Microglial clearance of dying neurons and associated debris is essential to maintain healthy neural networks in the central nervous system. In fact, the functions of microglia are regulated by various signaling molecules that are produced as neurons degenerate. Here, we show that the soluble CX3C chemokine fractalkine (sFKN), which is secreted from neurons that have been damaged by glutamate, promotes microglial phagocytosis of neuronal debris through release of milk fat globule-EGF factor 8, a mediator of apoptotic cell clearance. In addition, sFKN induces the expression of the antioxidant enzyme heme oxygenase-1 (HO-1) in microglia in the absence of neurotoxic molecule production, including NO, TNF, and glutamate. sFKN treatment of primary neuron-microglia co-cultures significantly attenuated glutamate-induced neuronal cell death. Using several specific MAPK inhibitors, we found that sFKN-induced heme oxygenase-1 expression was primarily mediated by activation of JNK and nuclear factor erythroid 2-related factor 2. These results suggest that sFKN secreted from glutamate-damaged neurons provides both phagocytotic and neuroprotective signals.