动力相关蛋白Drp1在羊瘙痒因子139A感染小鼠脑组织中变化的研究

目的 研究动力相关蛋白1(Drp1)在羊瘙痒因子139A感染的小鼠脑组织中的变化。方法 采用Western Blot方法检测Drp1在羊瘙痒因子139A感染的脑组织匀浆中的含量变化及其亚硝基化水平。采用组织免疫荧光方法检测Drp1在脑组织中的分布。结果 在羊瘙痒因子139A感染终末期小鼠脑组织匀浆中,Drp1总量略有降低。对感染不同时间点的动态分析结果显示Drp1含量呈逐渐降低趋势,终末期稍有回升。感染终末期脑组织中亚硝基化水平明显增高。组织免疫荧光显示正常和羊瘙痒因子139A感染终末期小鼠脑组织中,Drp1与神经元细胞存在共定位现象。结论 Drp1在小鼠中枢神经系统中主要分布于神经元细胞,在羊瘙痒因子139A感染的小鼠脑组织中,Drp1总量略有降低,另外亚硝基化水平明显增高,提示在羊瘙痒病感染的小鼠脑组织中,线粒体动力相关蛋白的表达量及翻译后修饰水平出现了变化,在感染过程中起着重要作用。     

核转录因子-B(p65)在羊瘙痒因子139A感染小鼠脑组织中变化的研究

目的 研究NF-B（p65）在羊瘙痒因子139A感染小鼠脑组织中的变化。方法 通过蛋白质免疫印迹法及免疫组织化学的方法检测p65在139A感染的脑组织均浆中的含量变化。利用免疫荧光方法检测p65在神经元及胶质细胞中的分布情况。结果 在139A感染终末期小鼠脑组织均浆中,p65含量下降,差异有统计学意义。对感染不同时间点的动态分析结果显示,p65含量呈先升高随后逐渐降低趋势。免疫组织化学显示,139A感染终末期小鼠大脑皮层p65含量明显下降。免疫组织荧光显示在正常和139A感染终末期小鼠脑组织中,p65与神经元细胞存在共定位现象。结论 NF-B（p65）在小鼠中枢神经系统中主要分布于神经元细胞,在139A感染的小鼠终末期脑组织中,p65总量明显下降,提示在139A感染的小鼠脑组织中,NF-B（p65）出现了变化,在疾病进程中发挥一定作用。     

CXCR7在碱烧伤小鼠角膜组织中的表达

目的:研究基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)的受体即人CXC趋化因子受体7(C-X-C chemokine receptor type 7,CXCR7)在碱烧伤小鼠角膜组织的表达,探讨CXCR7是否参与小鼠碱烧伤后角膜新生血管(corneal neovascularization,CNV)的形成。方法:采用在角膜鼻侧放置NaOH滤纸片40 s的方法构建BALB/c小鼠碱烧伤诱导CNV动物模型,通过real-time PCR、Western blotting法分别检测碱烧伤后3 d、7 d、14 d角膜组织中SDF-1、CXCR4、CXCR7和血管内皮生长因子(vascular endothelial growth factor,VEGF)的mRNA及蛋白的表达情况,免疫荧光法检测碱烧伤后14 d CXCR7蛋白的表达。结果:小鼠角膜新生血管在碱烧后14 d内持续生长。Real-time PCR结果显示,与正常对照组相比,碱烧伤后3d、7 d、14 d小鼠角膜组织中SDF-1、CXCR4、CXCR7及VEGF的mRNA表达均明显升高(P<0.05)。Western blotting结果显示,与正常组相比,碱烧伤后7 d组与14 d组中SDF-1、CXCR4、CXCR7及VEGF蛋白的相对表达量均升高(P<0.05),但7d组与14 d组差异无统计学意义。免疫荧光检测显示,正常小鼠角膜上皮层有CXCR7蛋白弱阳性蛋白表达;碱烧伤后14 d,角膜上皮层、基质层及新生血管CXCR7蛋白的表达升高。结论:CXCR7在碱烧伤后小鼠角膜组织中表达增加,提示其可能在碱烧伤后的CNV形成中发挥重要作用。     

CXCL12/CXCR4与子痫前期及其肝功能异常的相关性研究

目的 比较子痫前期患者和正常孕妇血清趋化因子CXCL12/CXCR4的水平,分析其与子痫前期孕妇肝功能异常的相关性.方法 选取2018年5月～2019年5月于安徽医科大学附属妇幼保健院住院行剖宫产分娩的子痫前期患者65例(PE组)和正常妊娠孕妇51例(对照组),将PE患者进一步分为轻度PE患者23例(轻度PE组)和重度...     

朊病毒感染细胞模型中趋化因子CXCL1变化特征研究

目的 探究CXCL1在朊病毒感染细胞模型SMB-S15中的变化特征,比较感染细胞内朊病毒含量变化对CXCL1水平变化的影响.方法 通过荧光定量PCR、蛋白免疫印迹和酶联免疫吸附试验(ELISA)对朊病毒感染细胞模型SMB-S15及其对照细胞SMB-PS、SMB-RES中CXCL1的转录及翻译水平进行分析.利用ELISA...     

CXCL11和CXCR3在宫颈癌患者中的表达及其与高危型人乳头状瘤病毒感染的关系

目的 探讨宫颈癌患者CXC趋化因子配体11(CXCL11)、CXC趋化因子受体-3(CXCR3)表达及其与高危型人乳头状瘤病毒(HR-HPV)感染的关系。方法 选取2019年5月至2022年5月该院诊治的宫颈癌患者75例为宫颈癌组,宫颈上皮内瘤变(CIN)患者78例为CIN组,宫颈炎患者52例为宫颈炎组。采用免疫组化法测定宫颈病变组织CXCL11、CXCR3表达,检测患者HR-HPV感染情况,分析CXCL11、CXCR3表达与HR-HPV感染的关系。结果 宫颈癌组宫颈病变组织CXCL11、CXCR3阳性表达率及HR-HPV阳性感染率分别为85.33%、80.00%、90.67%,高于宫颈炎组的15.38%、11.54%、13.46%及CIN组的52.56%、50.00%、62.82%,差异均有统计学意义(P<0.05);CIN组CXCL11、CXCR3阳性表达率及HR-HPV阳性感染率高于宫颈炎组(P<0.05)。宫颈癌患者宫颈病变组织CXCL11、CXCR3表达及HR-HPV感染与淋巴结转移、分化程度有关(P<0.05)。宫颈癌患者中HR-HPV感染共68例,其中单...     

HE4、CXCL12及其受体在卵巢癌中的表达及意义

目的探讨人附睾上皮分泌蛋白4(HE4)、CXC趋化因子配体12(CXCL12)及其受体[CXC趋化因子受体4(CXCR4)]在卵巢癌中的表达及意义。方法选取2016年3月至2018年3月在山东省胶州市妇幼保健院就诊的卵巢癌患者30例纳入卵巢癌组,另外选取在山东省胶州市妇幼保健院就诊的卵巢良性肿瘤患者30例纳入良性肿瘤组。取卵巢良性肿瘤患者手术切除组织标本(良性肿瘤组织)、卵巢癌患者手术切除癌组织标本(卵巢癌组织)以及距离癌组织5cm以上癌旁组织标本(癌旁组织),进行免疫组化标记以及革兰染色。采用酶联免疫吸附试验检测卵巢癌患者和卵巢良性肿瘤患者HE4、CXCL12及CXCR4水平,采用免疫组化染色观察各组织中HE4、CXCL12、CXCR4阳性表达情况。结果与良性肿瘤组相比,卵巢癌组患者的HE4、CXCL12及CXCR4水平升高(P0.05)。Ⅲ～Ⅳ期、低分化、有淋巴结转移的患者HE4、CXCL12及CXCR4的阳性率较高(P0.05)。HE4水平与CXCL12呈正相关,HE4水平与CXCR4呈正相关,CXCL12水平与CXCR4呈正相关。结论 HE4、CXCL12及CXCR4的异常表达参与了卵巢癌的发生、发展。该研究为HE4、CXCL12及CXCR4成为治疗卵巢癌的新靶点提供了理论依据。     

SDF-1/CXCR4在体外受精胚胎移植后输卵管妊娠患者输卵管中的表达及意义

目的 比较体外受精胚胎移植(IVF-ET)后输卵管妊娠和自然受孕输卵管妊娠患者基质细胞衍生因子-1(SDF-1)及CXC趋化因子受体4(CXCR4) mRNA表达的差异,以探讨其在IVF-ET后输卵管妊娠发生中所起的作用及机制.方法 分别选取2011年1月至2012年12月10例IVF-ET后输卵管妊娠和10例自然受孕输卵管妊娠患者,在腹腔镜手术的同时抽吸蜕膜、绒毛和输卵管黏膜组织,-70℃冷冻备存;采用荧光定量PCR的方法检测各组织中SDF-1及CXCR4mRNA表达.结果 两组患者胎盘绒毛、输卵管黏膜及子宫蜕膜组织的SDF-1及CXCR4mRNA表达量比较差异均无统计学意义(P均＞0.05);两组绒毛组织和输卵管黏膜组织均高表达CXCR4与SDF-1mRNA,其表达量显著高于各自的蜕膜组织(P均＜0.05).显示出受孕方式与输卵管妊娠发生率可能无关.结论 输卵管黏膜SDF-1和CXCR4表达亢进,子宫蜕膜SDF-1和CXCR4相对表达不足可能是IVF-ET后胚胎定植于输卵管的重要机制之一.     

羊瘙痒因子139A感染引起小鼠脑组织中的1-ACT表达增加

目的 探究1-ACT在羊瘙痒因子139A感染小鼠脑组织中的变化情况。方法 利用蛋白免疫印迹、免疫组织化学、间接免疫荧光及荧光共聚焦方法分析羊瘙痒因子139A感染小鼠脑组织中1-ACT表达的变化和分布特点。结果 蛋白免疫印迹方法显示在羊瘙痒因子139A感染小鼠终末期脑组织中1-ACT的含量较正常对照小鼠明显上调且随着潜伏期的延长而逐渐增加;免疫组织化学方法发现1-ACT主要分布于羊瘙痒因子139A感染小鼠的皮层、丘脑和小脑区域;间接免疫荧光实验显示羊瘙痒因子139A感染终末期小鼠脑组织中补体成分C3含量明显增加,同时荧光共聚焦实验表明1-ACT与补体成分C3存在明显的共定位现象。结论 羊瘙痒因子139A感染终末期小鼠脑组织中1-ACT含量明显增加。     

CXCR7/CXCL12在不同胃癌组织中的表达及与临床病理因素的相关性分析

目的分析趋化因子受体7(CXCR7)/趋化因子配体12(CXCL12)在不同胃癌组织中的表达及与临床病理因素的相关性。方法采用回顾性研究的方法,收集凉山州第一人民医院105例胃癌组织标本及其对应的正常胃黏膜组织标本。根据组织标本类型的不同,分为对照组(正常胃黏膜组织)、胃癌组、腹膜转移癌组、肝转移癌组、淋巴结转移癌组。采用免疫组织化学法检测各组标本中CXCR7与CXCL12蛋白的表达情况,进一步分析CXCR7和CXCL12蛋白表达与胃癌患者临床病理特征的相关性。结果 CXCR7蛋白表达在各组胃癌组织中的阳性率较对照组均明显升高(P 0. 05);而不同胃癌组织中CXCR7蛋白阳性表达率的比较,差异无统计学意义(P 0. 05)。CXCL12蛋白表达在各组胃癌组织中的阳性率较对照组均明显升高(P 0. 05);而不同胃癌组织中CXCL12蛋白阳性表达率的比较,差异无统计学意义(P 0. 05)。CXCR7与CXCL12蛋白表达与胃癌患者性别、年龄、肿瘤直径及肿瘤部位均无明显相关性(P 0. 05),而与肿瘤浸润深度、肿瘤组织形态、腹膜转移、肝脏转移、淋巴结转移、肿瘤侵袭淋巴管与血管均密切相关(P 0. 05)。结论 CXCR7与CXCL12在胃癌组织中的表达水平较正常胃黏膜组织明显升高,二者表达水平与胃癌发生、发展、增殖及迁移密切相关;随着胃癌进程的推进,CXCR7与CXCL12的表达水平可进一步增高,两者过高表达可能促使胃癌细胞的侵袭性及转移性更强,二者在胃癌发生、侵袭和转移的病理过程中可能具有协同作用,有望成为评估胃癌患者预后状况的潜在肿瘤标志物。     

Absence of autoantibodies against neurofilament proteins in the sera of scrapie infected mice

Autoantibodies against neurofilament proteins were not detected in any of the sera from the following scrapie infected mice: 19 mice infected with scrapie agent 139A in pre-clinical stage, 32 histologically confirmed scrapie mice and other 12 clinical scrapie mice infected with various strains. The test sera were assayed against acetone-fixed central neuron cultures from fetal mice by indirect immunofluorescence and immunoperoxidase techniques. The negative result suggests that autoantibodies against neurofilament proteins do not play a role in the pathogenesis of scrapie.     

趋化因子配体5及其受体趋化因子受体2在急性白血病髓外浸润中的表达和意义

目的观察趋化因子配体5(CXCL5)及其受体趋化因子受体2(CXCR2)在急性白血病(AL)髓外浸润(EI)中的表达,并探讨其临床意义。方法 60例初治成人AL患者,急性髓系白血病(AML)41例,急性淋巴系白血病(ALL)19例,骨穿取骨髓,ELISA法测定骨髓液中CXCL5含量,流式细胞术检测骨髓细胞CXCR2的相对荧光强度,统计分析其表达水平。结果 AL组、对照组的骨髓液CXCL5浓度[M(Q)]分别为3 173.93(1 392.22)pg/m L、349.88(178.67)pg/m L,差异有统计学意义(U=0.000,P0.05),CXCR2相对荧光强度分别为71.03(32.94)、19.68(8.90),差异有统计学意义(U=0.000,P0.05);伴有EI与无EI的骨髓CXCL5表达水平分别为3 687.81(885.05)pg/m L、2 305.32(781.85)pg/m L,差异有统计学意义(U=29,P0.05),骨髓细胞CXCR2相对荧光强度分别为80.72(21.11)、50.37(14.38),差异有统计学意义(U=16,P0.05);但AML组与ALL组的患者骨髓CXCL5/CXCR2表达差异无统计学意义(P0.05)。有效的化疗可以使AL患者获得完全缓解,使骨髓CXCL5、CXCR2表达水平明显下调;但在表达水平与完全缓解率方面,AML组与ALL组差异无统计学意义(P0.05)。AL患者骨髓液CXCL5浓度和骨髓细胞CXCR2的表达呈正相关(r=0.809,P=0.000);同时,骨髓CXCL5、CXCR2的表达与骨髓幼稚细胞百分比、外周血白细胞计数均呈正相关(P均0.05);与年龄、外周血小板计数均无明显相关性(P均0.05)。结论 AL患者的骨髓CXCL5、CXCR2表达均上调,在伴有EI的患者尤为明显,CXCL5、CXCR2轴的高表达可用于预测急性白血病的EI。     

盖尔森基兴诺卡菌GUH-2感染小鼠后细胞因子分泌特征研究

目的了解盖尔森基兴诺卡菌感染小鼠后细胞因子的分泌特征,进一步阐明其致病机制,为诺卡菌病的治疗提供理论基础。方法滴鼻感染BALB/c小鼠,采用平板计数法统计不同时间点小鼠肺上清及血清中的菌载量。 采用酶联免疫吸附试验（ELISA）检测在不同时间点肺上清及血清中多种细胞因子的浓度。 采用SPSS 23.0软件进行统计学分析。结果小鼠感染48 h后,98.35%的肺内盖尔森基兴诺卡菌被清除,在血清中没有检测到该菌。 在感染早期,肺组织中IL-2、IL-4、IL-6、IL-10、IL-12、TNF-α、IFN-γ的浓度均明显高于对照组,差异有统计学意义（P<0.01）,随着时间的推移,各细胞因子的浓度呈现逐渐下降的趋势。 在血清中,IL-6（6～24 h）、IFN-γ（12～24 h）与IL-10（48 h）浓度较高,与对照组相比差异有统计学意义（P<0.01）。IL-12（12～48 h）与IL-4（12 h）的表达受到抑制,与对照组相比差异有统计学意义（P<0.01）。 另外,在肺中,各细胞因子的浓度与菌载量之间符合线性相关,且均呈正相关。结论在小鼠感染盖尔森基兴诺卡菌的过程中,机体会分泌IL-6、TNF-α、IL-12、IL-2、IFN-γ、IL-10、IL-4等细胞因子,尤其是IL-6、IL-12、IFN-γ和IL-10浓度较高。 各细胞因子交互作用参与了清除盖尔森基兴诺卡菌感染的免疫效应机制。     

Role and implications of the CXCL12/CXCR4/CXCR7 axis in atherosclerosis: still a debate

Atherosclerosis is one of the leading causes of mortality and morbidity worldwide. Chemokines and their receptors are implicated in the pathogenesis of atherosclerosis. CXCL12 is a member of the chemokine family exerting a myriad role in atherosclerosis through its classical CXCR4 and atypical ACKR3 (CXCR7) receptors. The modulatory and regulatory functional spectrum of CXCL12/CXCR4/ACKR3 axis in atherosclerosis spans from proatherogenic, prothrombotic and proinflammatory to atheroprotective, plaque stabilizer and dyslipidemia rectifier. This diverse continuum is executed in a wide range of biological units including endothelial cells (ECs), progenitor cells, macrophages, monocytes, platelets, lymphocytes, neutrophils and vascular smooth muscle cells (VSMCs) through complex heterogeneous and homogenous coupling of CXCR4 and ACKR3 receptors, employing different downstream signalling pathways, which often cross-talk among themselves and with other signalling interactomes. Hence, a better understanding of this structural and functional heterogeneity and complex phenomenon involving CXCL12/CXCR4/ACKR3 axis in atherosclerosis would not only help in formulation of novel therapeutics, but also in elucidation of the CXCL12 ligand and its receptors, as possible diagnostic and prognostic biomarkers.

Key messages

1. The role of CXCL12 per se is proatherogenic in atherosclerosis development and progression.
2. The CXCL12 receptors, CXCR4 and ACKR3 perform both proatherogenic and athero-protective functions in various cell types
3. Due to functional heterogeneity and cross talk of CXCR4 and ACKR3 at receptor level and downstream pathways, regional boosting with specific temporal and spatial modulators of CXCL12, CXCR4 and ACKR3 need to be explored.

     

CXCL12-CXCR4/CXCR7趋化因子轴在肿瘤中的研究进展

近年来,多种研究表明CXCR4与CXCR7在肿瘤的发生发展过程中发挥着重要作用,CXCL12-CXCR4/CXCR7趋化因子轴与肿瘤生长和转移的关系引起人们的关注。本文对CXCL12-CXCR4/CXCR7在肿瘤中的表达、促进肿瘤增殖、侵袭转移及与肿瘤干细胞相关性等方面作一综述,提示CXCL12-CXCR4/CXCR7轴可成为抗肿瘤药物治疗的新靶点。     

Blockade of CXCL12/CXCR4 axis ameliorates murine experimental colitis

Recent studies indicate that the CXCL12/CXCR4 interaction is involved in several inflammatory conditions. However, it is unclear whether this interaction has a role in the pathophysiology of inflammatory bowel disease (IBD). We investigated the significance of this interaction in patients with IBD and in mice with dextran sulfate sodium (DSS)-induced colitis and the effect of a CXCR4 antagonist on experimental colitis. First, we measured CXCR4 expression on peripheral T cells in patients with IBD. Furthermore, we investigated CXCR4 expression on leukocytes and CXCL12 expression in the colonic tissue of mice with DSS-induced colitis, and we evaluated the effects of a CXCR4 antagonist on DSS-induced colitis and colonic inflammation of interleukin (IL)-10 knockout (KO) mice. Colonic inflammation was assessed both clinically and histologically. Cytokine production from mesenteric lymph node cells was also examined. CXCR4 expression on peripheral T cells was significantly higher in patients with active ulcerative colitis (UC) compared with normal controls, and CXCR4 expression levels of UC patients correlated with disease activity. Both CXCR4 expression on leukocytes and CXCL12 expression in colonic tissue were significantly increased in mice with DSS-induced colitis. Administration of a CXCR4 antagonist ameliorated colonic inflammation in DSS-induced colitis and IL-10 KO mice. CXCR4 antagonist reduced tumor necrosis factor-alpha and interferon-gamma production from mesenteric lymph node cells, whereas it did not affect IL-10 production. The percentage of mesenteric Foxp3+CD25+ T cells in DSS-induced colitis was not affected by CXCR4 antagonist. These results suggest that blockade of this chemokine axis might have potential as a therapeutic target for the treatment of IBD.     

Pharmacological characterization of Sch527123, a potent allosteric CXCR1/CXCR2 antagonist

In neutrophils, growth-related protein-alpha (CXCL1) and interleukin-8 (CXCL8), are potent chemoattractants (Cytokine 14:27-36, 2001; Biochemistry 42:2874-2886, 2003) and can stimulate myeloperoxidase release via activation of the G protein-coupled receptors CXCR1 and CXCR2. The role of CXCR1 and CXCR2 in the pathogenesis of inflammatory responses has encouraged the development of small molecule antagonists for these receptors. The data presented herein describe the pharmacology of 2-hydroxy-N,N-dimethyl-3-{2-[[(R)-1-(5-methyl-furan-2-yl)-propyl]amino]-3,4-dioxo-cyclobut-1-enylamino}-benzamide (Sch527123), a novel antagonist of both CXCR1 and CXCR2. Sch527123 inhibited chemokine binding to (and activation of) these receptors in an insurmountable manner and, as such, is categorized as an allosteric antagonist. Sch527123 inhibited neutrophil chemotaxis and myeloperoxidase release in response to CXCL1 and CXCL8 but had no effect on the response of these cells to C5a or formyl-methionyl-leucyl-phenylalanine. The pharmacological specificity of Sch527123 was confirmed by testing in a diversity profile against a panel of enzymes, channels, and receptors. To measure compound affinity, we characterized [(3)H]Sch527123 in both equilibrium and nonequilibrium binding analyses. Sch527123 binding to CXCR1 and CXCR2 was both saturable and reversible. Although Sch527123 bound to CXCR1 with good affinity (K(d) = 3.9 +/- 0.3 nM), the compound is CXCR2-selective (K(d) = 0.049 +/- 0.004 nM). Taken together, our data show that Sch527123 represents a novel, potent, and specific CXCR2 antagonist with potential therapeutic utility in a variety of inflammatory conditions.     

Chemokine contribution to neuropathic pain: Respective induction of CXCL1 and CXCR2 in spinal cord astrocytes and neurons

Recent studies have indicated an important role of chemokines such as CCL2 in the development of chronic pain. However, the distinct roles of different chemokines in the development and maintenance of neuropathic pain and in their interactions with neurons have not been clearly elucidated. We found that spinal nerve ligation (SNL) not only induced persistent neuropathic pain symptoms, including mechanical allodynia and heat hyperalgesia, but also produced sustained CXCL1 upregulation in the spinal cord. Double staining of immunofluorescence and in situ hybridization revealed that CXCL1 was primarily induced in spinal astrocytes. In cultured astrocytes, tumor necrosis factor-α induced robust CXCL1 expression via the activation of the c-jun N-terminal kinase. Intrathecal administration of CXCL1 neutralizing antibody transiently reduced SNL-induced pain hypersensitivity, suggesting an essential role of CXCL1 in neuropathic pain sensitization. In particular, intraspinal delivery of CXCL1 shRNA lentiviral vectors, either before or after SNL, persistently attenuated SNL-induced pain hypersensitivity. Spinal application of CXCL1 not only elicited pain hypersensitivity but also induced rapid neuronal activation, as indicated by the expression of phosphorylated extracellular signal-regulated kinase and cAMP response element binding protein, and c-Fos in spinal cord neurons. Interestingly, CXCR2, the primary receptor of CXCL1, was upregulated in dorsal horn neurons after SNL, and the CXCR2 antagonist SB225002 completely blocked the CXCL1-induced heat hyperalgesia. SB225002 also attenuated SNL-induced pain hypersensitivity. Collectively, our results have demonstrated a novel form of chemokine-mediated glial-neuronal interaction in the spinal cord that can drive neuropathic pain. Inhibition of the CXCL1-CXCR2 signaling may offer a new therapy for neuropathic pain management.