非小细胞肺癌中CCR7和CD62L表达及与淋巴结转移的关系

目的探讨CC趋化因子受体7(CCR7)和L-选择素(CD62L)在非小细胞肺癌(NSCLC)组织中的表达及其与淋巴结转移的关系。方法采用免疫组化法检测80份NSCLC癌组织和20份癌旁正常肺组织标本中CCR7和CD62L表达,比较二者在淋巴结转移者与无淋巴结转移者中的阳性表达率。结果 NSCLC癌组织中CCR7和CD62L表达阳性率均明显高于癌旁正常肺组织(P均<0.05),且CCR7表达与CD62L表达有明显相关性(P<0.05)。有淋巴结转移者CCR7和CD62L表达阳性率均明显高于无淋巴结转移者(P均<0.05)。结论 NSCLC癌组织中CCR7和CD62L表达均上调,二者可能共同参与了NSCLC的淋巴结转移过程。     

Correlation between circulating fibrocytes, and activity and progression of interstitial lung diseases

Background and objective: Interstitial lung diseases (ILD) are characterized by progressive interstitial pulmonary fibrosis and a decline in lung function. Fibrocytes are bone marrow‐derived mesenchymal progenitor cells that may play a role in the pathogenesis of pulmonary fibrosis. Circulating fibrocyte numbers have been correlated with the prognosis of patients with idiopathic pulmonary fibrosis. The aim of the present study was to evaluate the relationship between circulating fibrocytes, and parameters of disease activity and progression in several groups of patients with ILD. Methods: The study population comprised 41 patients with ILD and seven healthy control subjects. Circulating CD45⁺ collagen‐I⁺ fibrocytes were evaluated by flow cytometry. Results: The number of circulating fibrocytes was significantly increased in all patients with ILD and particularly in patients with idiopathic interstitial pneumonitis and interstitial pneumonitis associated with collagen vascular disease as compared with healthy control subjects. The numbers of circulating fibrocytes were significantly correlated with pulmonary function test parameters and with serum levels of sialylated carbohydrate antigen, a marker of disease activity. Temporal changes in circulating fibrocyte numbers were evaluated in two patients, and the results suggested that these changes correlated with the activity of ILD. Conclusions: The results from this study provide further evidence for the role of circulating fibrocytes in fibrotic lung diseases.     

miR-106调控CC趋化因子配体2对增生型糖尿病视网膜病变中人视网膜微血管内皮细胞增殖、血管生成和炎症反应的影响

目的　探讨miR-106调控CC趋化因子配体2（CCL2）对增生型糖尿病视网膜病变（PDR）中人视网膜微血管内皮细胞（HRMEC）增殖、血管生成、炎症反应的影响。方法　GEO数据库筛选PDR中差异表达的miRNAs。高糖（25.0 mmol·L^-1葡萄糖,HG）诱导HRMEC建立PDR细胞模型,qRT-PCR检测miR-106和CCL2在正常葡萄糖（5.5 mmol·L^-1葡萄糖,NG）和HG培养条件下HRMEC中的表达。将PDR患者的血清外泌体做miR-106过表达处理后与HG处理的HRMEC共培养,同时干预HRMEC中CCL2的表达以探讨血清外泌体miR-106和CCL2在PDR中的功能。采用MTT法、小管形成实验和ELISA法分别检测各组HRMEC增殖、血管生成和炎症因子的表达。双荧光素酶报告实验用以验证miR-106和CCL2的靶向关系。结果　与NG组细胞中miR-106表达（1.04±0.10）、CCL2表达（1.02±0.09）相比,HG培养的HRMEC中miR-106表达（0.68±0.06）降低,CCL2表达（1.38±0.11）升高（均为P<0.05）。PDR患者血清外泌体中miR-106表达较NDR患者血清外泌体中表达降低（P<0.05）。与HG+PDR-exo+miR-NC组相比,HG+PDR-exo+miR-106 mimic组HRMEC活力降低,血管生成被抑制,细胞上清液中TNF-α、IL-1β和IL-6水平降低,而SOD、CAT和GSH水平升高（均为P<0.05）。双荧光素酶报告实验证实了CCL2是miR-106的一个靶点。与HG+PDR-exo+miR-106 mimic+oe-NC组相比,HG+PDR-exo+miR-106 mimic+oe-CCL2组HRMEC活力提高,血管生成被诱导,TNF-α、IL-1β和IL-6水平升高,而SOD、CAT和GSH水平降低（均为P<0.05）。结论　血清外泌体miR-106能够抑制CCL2表达,进而对PDR中的HRMEC增殖、血管生成和炎症反应起抑制作用。     

电针联合脂肪源性干细胞移植对脑缺血/再灌注大鼠SDF-1和CXCR4表达的影响

目的探讨电针联合脂肪源性干细胞(ADSC)移植对大鼠缺血/再灌注损伤后趋化因子SDF-1及其受体CXCR4的影响。方法成年大鼠随机分为模型组、电针组、ADSC移植组、电针+ADSC组。线栓法制作大鼠大脑中动脉缺血(MCAO)2h再灌注模型,电针组取大椎穴和内关穴行电针治疗。ADSC组尾静脉注射PKH26标记的ADSC细胞悬液,电针+ADSC组联合电针和AD-SC移植治疗。缺血7d后行神经功能损害评分(NSS),采用Western bolt法及实时荧光定量PCR检测海马区SDF-1、CXCR4蛋白和mRNA表达。结果电针+ADSC组海马区PKH-26标记的细胞个数高于ADSC组(P<0.05)。与模型组比较,电针组、ADSC组和电针+ADSC组NSS评分均显著降低,海马区SDF-1、CXCR4蛋白和mRNA表达增加(P<0.05)。其中电针+ADSC组较电针组和ADSC组NSS评分显著降低,而SDF-1、CXCR4蛋白和mRNA表达显著增加(P<0.05)。结论电针联合脂肪源性干细胞移植促进脑缺血再灌注大鼠神经功能恢复作用优于单独治疗组,其机制可能与电针上调脑海马区SDF-1和CXCR4表达,促进移植的ADSC迁移和存活有关。     

MCP-1参与缬沙坦对阿霉素心脏毒性的预保护作用

目的　探讨阿霉素诱导SD大鼠心肌损伤模型中单核细胞趋化蛋白-1（MCP-1）蛋白表达及其潜在的干预机制。方法　选择8周龄雄性SPF级SD大鼠30只,按照随机数字表法分为3组：对照组（Con组,生理盐水灌胃＋腹腔注射）、阿霉素组（Dox组,1.25 mL/kg阿霉素腹腔注射+2 mL/kg生理盐水灌胃）及缬沙坦+阿霉素组（Val+Dox组,1.25 mL/kg阿霉素腹腔注射+2 mL/kg缬沙坦灌胃）各10只,连续干预6周。10周时超声心动图检测心功能变化,留取大鼠心肌组织,电镜观察心肌超微结构变化,采用蛋白芯片技术检测各组血清MCP-1、调节激活正常T细胞表达和分泌因子（RANTES）及次级淋巴组织趋化因子（6Ckine）蛋白含量,采用Western blot检测各组心肌组织MCP-1蛋白表达。结果　超声结果显示,与Con组相比,Dox组和Val+Dox组左心室射血分数（LVEF）、左心室短轴缩短率（LVFS）明显降低,左心室收缩末期内径（LVESD）、左心室舒张末期内径（LVEDD）明显升高（P＜0.05）。与Dox组相比,Val+Dox组LVEF、LVFS明显升高,LVESD、LVEDD明显降低（P＜0.05）。电镜下可见Con组心肌细胞结构清晰;Dox组心肌结构模糊,心肌溶解,可见自噬小体;Val+Dox组心肌结构模糊不清,线粒体体积增大、大小不等,但结构完整。与Con组相比,Dox组血清MCP-1、RANTES、6Ckine蛋白分别上调1.87倍、1.40倍和1.26倍（P＜0.05）。Western blot结果显示,Dox组与Con组相比,心肌组织MCP-1蛋白表达显著升高（P＜0.05）;Val+Dox组与Dox组相比,心肌组织MCP-1蛋白表达显著降低（P＜0.05）。结论　MCP-1蛋白可能参与阿霉素诱导的心肌损伤,缬沙坦可能通过下调MCP-1蛋白表达减轻心肌损伤。     

Designing small molecule CXCR3 antagonists

Introduction: By virtue of its specificity for chemokines induced in Th1-associated pathologies, CXCR3 has attracted considerable attention as a target for therapeutic intervention. Several pharmacologically distinct small molecules with in vitro and in vivo potency have been described in the literature, although to date, none have shown efficacy in clinical trials.

Areas covered: In this article, the author outlines the rationale for targeting CXCR3 and discusses the potential pitfalls in targeting receptors in poorly understood areas of chemokine biology. Furthermore, they cover emerging therapeutic areas outside of the ‘traditional’ Th1 arena in which CXCR3 antagonists may ultimately bear fruit. Finally, they discuss the design of recently discovered small molecules targeting CXCR3.

Expert opinion: CXCR3 and its ligands appear to play roles in a multitude of diverse diseases in humans. In vitro studies suggest that CXCR3 is inherently ‘druggable’ and that potent, efficacious small molecules targeting CXCR3 antagonists will find a clinical niche. However, the well-trodden path to failure of small molecule chemokine receptor antagonists in clinical trials suggests that a cautious approach should be undertaken. Ideally, unequivocal evidence elucidating the precise role of CXCR3 should be obtained before targeting the receptor in a particular disease cohort.     

Ly6Chigh Monocytes Protect against Kidney Damage during Sepsis via a CX3CR1-Dependent Adhesion Mechanism

Monocytes have a crucial role in both proinflammatory and anti-inflammatory phenomena occurring during sepsis. Monocyte recruitment and activation are orchestrated by the chemokine receptors CX3CR1 and CCR2 and their cognate ligands. However, little is known about the roles of these cells and chemokines during the acute phase of inflammation in sepsis. Using intravital microscopy in a murine model of polymicrobial sepsis, we showed that inflammatory Ly6C^high monocytes infiltrated kidneys, exhibited altered motility, and adhered strongly to the renal vascular wall in a chemokine receptor CX3CR1-dependent manner. Adoptive transfer of Cx3cr1-proficient monocyte-enriched bone marrow cells into septic Cx3cr1-depleted mice prevented kidney damage and promoted mouse survival. Modulation of CX3CR1 activation in septic mice controlled monocyte adhesion, regulated proinflammatory and anti-inflammatory cytokine expression, and was associated with the extent of kidney lesions such that the number of lesions decreased when CX3CR1 activity increased. Consistent with these results, the pro-adhesive I249 CX3CR1 allele in humans was associated with a lower incidence of AKI in patients with sepsis. These data show that inflammatory monocytes have a protective effect during sepsis via a CX3CR1-dependent adhesion mechanism. This receptor might be a new therapeutic target for kidney injury during sepsis.