趋化因子CX3CL1及其受体CX3CR1在神经病理性痛中的调节作用

正神经病理性痛是一种复杂的疾病,现已成为困扰全球的健康问题。其发生原因多种多样,包括:病毒感染、肿瘤、外周或中枢神经系统损伤以及糖尿病等伴有周围神经损害的疾病,甚至某些可以导致神经损伤的药物。神经病理性痛是由躯体感觉神经系统的损伤或疾病而直接造成的疼痛(pain arising as a direct consequence of a lesion or disease af fecting the somatosensory system)~[1],最常见的症状一般为痛     

结肠癌患者CX3CL1的表达情况与术后预后的相关性分析

目的探讨结肠癌组织中CX3CL1的表达情况、临床意义及其与术后预后分析的相关性。方法应用免疫组织化学法检测104例结肠癌病人的石蜡切片中CX3CL1的表达情况,用χ2检验来分析CX3CL1的临床意义,用Kaplan-Meier方法及建立Cox回归模型来分析CX3CL1表达情况与结肠癌术后预后的相关性。结果 CX3CL1高表达组占所有结肠癌病人的61.54%。CX3CL1的表达与年龄、性别、肿瘤大小、肿瘤位置、Dukes分期、组织分化程度无明显相关（P〉0.05）,但与淋巴结转移相关（P〈0.05）。CX3CL1高表达组（n=64）比低表达组（n=40）有更长的生存期（P=0.0096）。Cox回归模型分析表明,CX3CL1表达可作为判断结肠癌患者术后生存和预后的独立指标（RR=2.4;P=0.0135）。结论 CX3CL1可作为结肠癌的独立预后因素,结肠癌中CX3CL1高表达提示更好的预后。     

糖皮质激素对BXSB狼疮小鼠肾组织fractalkine/CX3CR1表达的影响

目的：研究BXSB狼疮小鼠肾组织趋化因子fractalkine及其受体CX3CR1的表达以及给予泼尼松治疗后的改变,探讨两者在狼疮肾炎发病机制中的可能作用。方法：12 周龄雄性BXSB狼疮小鼠随机分成泼尼松治疗组（n=6）和实验对照组（n=6）;另取同周龄雄性C57BL/6J小鼠6只作为正常对照组。正常对照组和实验对照组小鼠每天给予0.5 mL生理盐水灌胃;泼尼松治疗组小鼠每天给予0.18 mg/20 g BW的泼尼松溶于0.5 mL生理盐水灌胃。持续10周结束实验。应用逆转录-聚合酶链反应（RT-PCR）及Western印迹检测小鼠肾组织fractalkine和CX3CR1 mRNA和蛋白的表达,并检测小鼠实验室指标以及肾脏组织病理学的变化。结果： BXSB狼疮小鼠肾组织fractalkine以及CX3CR1 mRNA和蛋白表达均较C57BL/6J小鼠明显增高,而经过泼尼松治疗后的BXSB小鼠两者的表达均较未治疗组（实验对照组）明显下降,同时伴有血清免疫球蛋白G（IgG）、IgM、血清抗双链脱氧核糖核酸（dsDNA）抗体水平以及血尿素氮（BUN）、血肌酐（SCr）水平的明显改善,尿蛋白减少;肾小球内免疫复合物沉积和肾脏组织病理学改变亦显著减轻。结论： 实验结果提示fractalkine/CX3CR1可能参与了小鼠狼疮肾炎的发病机制,且糖皮质激素可能通过抑制肾脏fractalkine的表达而发挥其治疗效应。     

764—3对大鼠低氧性肺动脉高压的作用

将大鼠置常压低氧舱(10％O_2～90％N_2)观察肺动脉压、右心室重及右心室功能的动态变化过程及764-3对其影响。低氧3天时除肺动脉压升高外上述其它指标无明显改变。自低氧7天时起,右心室重量及右心室功能也显著高于对照组,并持续至观察的21天。低氧21天后再吸常氧14天,上述变化基本恢复正常。764-3处理可显著缓解低氧所致的上述变化。结果提示低氧所致的变化在复氧后一定时期内可自然消退,764-3对低氧性肺动脉高压及右心室肥厚有明显的保护作用,作用机理有待进一步研究。     

肺动脉高压患者血浆Apelin的变化及意义

目的:观察新发现的小分子活性肽Apelin在肺动脉高压(PH)病人血浆中的变化,探讨其在肺动脉高压中可能的作用。方法:采用放射免疫分析测定32例PH患者和30例健康人(NC组)血浆Apelin水平;PH组另行血气分析及测定肺动脉收缩压(PASP)。结果:①血浆Apelin水平(pg/m l);PH组(59±31)较NC组(80±30)低26.3%(P<0.01);②PH组动脉血气分析:PaO2(64±13)mmHg,PaCO2(55±13)mmHg,pH(7.37±0.05)。PASP:(47±16)mmHg。③相关分析:血浆Apelin水平与PASP呈显著负相关(r=-0.65,P<0.01)。结论:肺动脉高压患者血浆Apelin水平降低,其生成减少和/或代谢异常可能是导致肺循环稳态失衡的重要因素之一。     

肺动脉高压患者肺组织S1PR1/2/3的表达

目的探讨肺动脉高压患者肺组织1-磷酸鞘氨醇受体1/2/3(sphingosine-1-phosphate receptor1/2/3,S1PR1/2/3)的表达变化,为肺动脉高压发病机制的深入研究奠定基础。方法人的肺组织来源于黑龙江省医院,其中对照组取自右下肺叶球形性变实施肺叶切除患者的肺组织,病变组取自患有特发性肺动脉高压尸检患者的肺组织,应用免疫印迹及实时定量PCR技术检测S1PR1/2/3的表达变化。结果原发性肺动脉高压患者肺组织S1PR1/2/3蛋白的光密度值分别为2.00±0.03,0.86±0.13,0.94±0.03,m RNA的绝对定量值分别为3.90±0.42,2.50±0.37,92.08±11.26,较对照组蛋白的光密度值(0.88±0.29,0.27±0.07,0.54±0.06)及m RNA的绝对定量值(1.44±0.33,1.12±0.15,1.63±0.56)明显增高(P0.01)。结论原发性肺动脉高压患者肺组织S1PR1/2/3的表达上调,提示S1PR1/2/3在肺动脉高压发病过程中可能发挥了重要的作用。     

肺动脉高压尸检病例的肺动脉形态计量及胶原变化的观察

为探讨原发性肺动脉高压病例肺动脉的病理分级与血液动力学变化及Ⅰ、Ⅳ型胶原蛋白变化的关系，收集了我院尸检有右心导管资料的９例病例（丛状肺动脉病６例，弥漫性肺间质纤维化２例，慢性肺栓塞及血栓形成１例）作为肺动脉高压组。将肺动脉高压性病变分４级：Ⅰ级仅出现肺小动脉中层肥厚；Ⅱ级为Ⅰ级加内膜细胞增生；Ⅲ级为Ⅰ级加内膜纤维化管腔狭窄及（或）血栓阻塞；Ⅳ级为Ⅲ级加丛状病变，无或有坏死性动脉炎。另外用免疫酶标ＰＡＰ染色法进行抗Ⅰ型、Ⅳ型胶原抗体检测。结果如下：（１）正常７例对照组肺小动脉中层平均厚为外径６．１％，肺小动脉密度平均为２２．６支／ｃｍ２；肺动脉高压组肺动脉中层平均厚为外径的２６．２％，密度平均值４４．６支／ｃｍ２，两组分别经ｔ检验，Ｐ值均小于０．０１。提示肺动脉高压可引起肺动脉中层肥厚及肺小动脉密度增加。（２）统计学分析显示，肺动脉压力与病理分级呈正相关关系（ｒ＝０．６８，Ｐ＜０．０１）。（３）Ⅰ型胶原变化与病程有关，在肺动脉高压晚期不可逆性病变中Ⅰ型胶原增加；而Ⅳ型胶原增加反映为可逆性病变阶段。     

大鼠缺氧性肺动脉高压过程中缺氧诱导因子1α和血红素氧合酶1的变化

目的：观察缺氧大鼠肺小血管壁缺氧诱导因子1α（HIF-1α）和血红素氧合酶1（HO-1）基因表达。方法： 40只雄性Wistar大鼠随机分成缺氧0、3、7、14和21 d组。测平均肺动脉压（mPAP），血管形态学指标，右室肥大指数(RVHI)，HO-1活性和HIF-1α, HO-1 基因表达。结果： 缺氧7 d mPAP高于对照组，缺氧14 d达到高峰并维持于此水平。肺血管重塑，RVHI改变在缺氧14 d后出现。HIF-1α蛋白在对照组表达不明显，各缺氧组血管内膜均为阳性。在中膜，HIF-1α蛋白缺氧3 d开始增高，第7 d达到高峰，14 d和21 d后下降，HIF-1α mRNA缺氧14 d增高, 此后维持于高水平。HO-1蛋白在缺氧7 d后增高，14 d后达到高水平，并持续于高水平。HO-1 mRNA缺氧3 d增高，7 d达高峰，之后下降。结论： HIF-1α和HO-1 均在大鼠缺氧性肺动脉高压的发病机制中发挥作用，且HIF-1α与HO-1基因表达可能有相互调控。     

miR-183靶向CX3CL1基因调控肝癌细胞MICA/B表达和增加NK细胞对肝癌细胞杀伤作用

目的探讨miR-183对肝癌细胞MHC-Ⅰ类相关蛋白A/B(MICA/B)表达和自然杀伤(NK)细胞杀伤作用影响及其分子机制。方法实验设置miR-con组、miR-183组、anti-miR-con组、anti-miR-183组、pcDNA组、pcDNA-CX3CL1组、antimiR-183+si-con组、anti-miR-183+CX3CL1组、对照(NC)组。实时荧光定量PCR(RT-qPCR)检测miR-183和CX3CL1 m RNA表达水平;蛋白质印迹(Western blot)法检测CX3CL1、MICA和MICB蛋白表达;流式细胞术检测肝癌细胞表面MICA/B的表达;NK细胞与肝癌细胞共培养后,细胞计数试剂盒8(CCK-8)法检测NK细胞对肝癌细胞的杀伤作用;酶联免疫吸附法(ELISA)法检测肝癌细胞与NK细胞共培养后培养液中肿瘤坏死因子-α(TNF-α)和干扰素γ(IFN-γ)水平;荧光素酶报告实验检测miR-183和CX3CL1的靶向关系。结果肝癌组织和肝癌细胞中miR-183高表达,CX3CL1、MICA和MICB低表达。干扰miR-183表达和过表达CX3CL1,肝癌细胞中MICA、MICB阳性表达率显著升高,与NK细胞共培养后TNF-α和IFN-γ水平显著升高,NK细胞对肝癌细胞的杀伤率显著升高(P<0.05)。miR-183靶向调控CX3CL1,沉默CX3CL1逆转了干扰miR-183对肝癌细胞MICA/B表达和NK细胞杀伤作用的影响。结论抑制表达miR-183可增加肝癌细胞MICA/B表达,增加NK细胞对肝癌细胞的杀伤作用,其机制可能与CX3CL1有关。     

NF-κB通路对类风湿关节炎成纤维样滑膜细胞中CX3CL1表达的影响

目的观察NF-κB通路对类风湿关节炎成纤维样滑膜细胞(RA-FLS)中CX3CL1表达的影响。方法用100μM NF-κB拮抗剂(PDTC)刺激体外培养的RA-FLS,用RT-PCR检测刺激不同时间后RA-FLS中CX3CL1 mRNA表达。结果 RA-FLS中表达CX3CL1;PDTC作用18h后,RA-FLS自身CX3CL1 mRNA表达量下降(P0.001)。结论 CX3CL1 mRNA的表达可能受到NF-κB通路的调节。     

CX3CL1 in allergic diseases: not just a chemotactic molecule

Allergic asthma and atopic dermatitis (AD) are two allergic diseases that are primarily driven by the activation of T helper (Th)2 cells. Th2 cells produce cytokines that directly contribute to the symptoms of these diseases. The recruitment and maintenance of Th2 cells into the target tissues are two key events in the pathogenesis of allergic asthma and AD. While migration is mediated by both chemokines and lipid mediators such as leukotrienes and prostaglandins, very little is known about the molecules involved in lymphocyte survival and maintenance in inflamed tissues. However, chemokines could also play a role in this phenomenon. An example of this could be illustrated by CX3CL1, also known as fractalkine. CX3CL1 is a chemokine that is upregulated in some inflammatory diseases including allergic pathologies and that was recently demonstrated to provide a survival signal upon binding to its unique receptor CX3CR1.     

Expression of CX3CL1 (fractalkine) in mice with endothelial-target rickettsial infection of the spotted-fever group

Fractalkine (CX3CL1) is a chemokine expressed mainly by endothelial cells, which are the major cellular targets of rickettsiae. We used immunohistochemistry to investigate the normal expression of CX3CL1 in mice and the kinetics of expression of this chemokine throughout the course of lethal and sublethal rickettsial infections in a mouse model of spotted-fever group rickettsioses. The peak of expression of fractalkine on day 3 of infection coincided with the time of infiltration of macrophages into infected tissues and preceded the peak of rickettsial content in tissues.     

Type I interferon induces CX3CL1 (fractalkine) and CCL5 (RANTES) production in human pulmonary vascular endothelial cells

Type I interferon (IFN) medications cause various adverse reactions, including vascular diseases. Although an association between chemokines and vascular diseases has also been reported, the relationship between type I IFN and chemokines in vascular endothelial cells (VEC) remains unclear. To provide clues to pathogenesis of the diseases, we analysed the effects of type I IFN on chemokine production in human VEC. Type I IFN induced higher CX3CL1 (fractalkine) mRNA expression and protein secretion in pulmonary arterial VEC than in umbilical vein VEC. Type I IFN also induced CCL5 [regulated upon activation normal T cell expressed and secreted (RANTES)] production in VEC, especially in lung micro-VEC. IFN-β induced much higher chemokine production than IFN-α, and Janus protein tyrosine kinase (JAK) inhibitor I prevented type I IFN-induced chemokine secretion. Type I IFN-induced chemokines may be involved in the pathophysiology of pulmonary vascular diseases, and the JAK inhibitor may serve as a therapeutic option for these diseases.     

The impact of chemokine receptor CX3CR1 deficiency during respiratory infections with Mycobacterium tuberculosis or Francisella tularensis

Recruitment of immune cells to infection sites is a critical component of the host response to pathogens. This process is facilitated partly through interactions of chemokines with cognate receptors. Here, we examine the importance of fractalkine (CX3CL1) receptor, CX3CR1, which regulates function and trafficking of macrophages and dendritic cells, in the host''s ability to control respiratory infections with Mycobacterium tuberculosis or Francisella tularensis. Following low-dose aerosol challenge with M. tuberculosis, CX3CR1^−/− mice were no more susceptible to infection than wild-type C57BL/6 mice as measured by organ burden and survival time. Similarly, following inhalation of F. tularensis, CX3CR1^−/− mice displayed similar organ burdens to wild-type mice. CX3CR1^−/− mice had increased recruitment of monocytes and neutrophils in the lung; however, this did not result in increased abundance of infected monocytes or neutrophils. We conclude that CX3CR1-deficiency affects immune-cell recruitment; however, loss of CX3CR1 alone does not render the host more susceptible to M. tuberculosis or F. tularensis.