银屑病患者中性粒细胞中CXC型趋化因子受体CXCR1、CXCR2 mRNA的表达

目的 了解银屑病患者外周血中性粒细胞中CXC型趋化因子受体CXCR1及CXCR2的表达情况。方法 应用逆转录-聚合酶链反应(RT-PCR)法检测了进行期斑块状银屑病患者及健康人对照各30例(其中治疗后患者14例)外周血中性粒细胞中CXCR1及CXCR2mRNA的表达,并将结果与患者皮损面积及严重程度指数(PASI)进行了相关性分析。结果 斑块状银屑病患者外周血中性粒细胞中CXCR1及CXCR2mRNA表达水平分别为1.30±1.18和1.62±0.97,明显高于正常人对照(分别为0.56±0.36、0.74±0.58,P<0.01),并随治疗后患者PASI评分的下降而降低,治疗后CXCR1、CXCR2分别为0.49±0.34、0.51±0.51,(P<0.01),银屑病患者外周血中性粒细胞中CXCR2mRNA水平高于CXCR1mRNA水平,二者均与PASI呈显著正相关。CXCR1:r=0.60,P<0.001;CXCR2:r=0.84,P<0.001。结论 银屑病患者外周血中性粒细胞中增高的CXCR1与CXCR2可能与白细胞向皮损部位的移行和聚集有关,CXCR1及CXCR2参与了银屑病的发病机制。     

CX3C型趋化因子受体CX3CR1 mRNA在银屑病患者中的表达

目的:测定趋化因子受体CX3CR1 mRNA在银屑病患者中的表达水平及其与PASI之间的关系。方法:应用RT-PCR法检测了33例斑块状银屑病患者皮损处、非皮损处皮肤和淋巴细胞及其中16例治疗后患者淋巴细胞中CX3CR1 mRNA的表达水平,设30例健康人为对照组;将检测结果与PASI及皮损处与淋巴细胞中该受体的表达水平分别进行相关性分析。结果:CX3CR1mRNA表达水平:在银屑病治疗前与治疗后外周血淋巴细胞中分别为1.64±1.19与2.15±1.30,均明显高于对照组(0.72±0.72,P<0.001);在银屑病皮损处表皮中为1.33±0.40,明显高于对照组(0.89±0.26,P<0.001)及非皮损处皮肤(0.94±0.45,P<0.01);在银屑病皮损处真皮中为1.59±0.76,明显高于对照组(0.89±0.41,P<0.001)及非皮损处皮肤(1.21±0.53,P<0.01),并与PASI(r=0.601,P<0.001)及淋巴细胞中该受体的表达水平之间呈正相关(r=0.351,P<0.05),非皮损处真皮中CX3CR1 mRNA表达水平也高于对照组(P<0.05)。结论:CX3CR1可能主要通过参与活化、趋化淋巴细胞而在银屑病的发病机制及病理状态的维持中起到重要作用。     

窄谱中波紫外线对银屑病患者CCR2 mRNA表达的影响

目的探讨窄谱中波紫外线(NB-UVB)治疗对银屑病患者趋化因子受体CCR2 mRNA的影响。方法应用逆转录-聚合酶链反应(RT-PCR)法检测36例斑块型银屑病患者NB-UVB治疗前、后皮损部位、外周血淋巴细胞中CCR2 mRNA的表达水平,30例正常人设为对照组,将检测结果与银屑病皮损面积、严重程度指数(PASI)及外周血白细胞中该受体的表达水平与皮损部位的表达水平间的相关性分别进行分析。结果CCR2 mRNA表达水平:银屑病患者治疗前淋巴细胞中为1.87±0.65,明显高于正常人对照组(0.31±0.19,P<0.001)及同组患者治疗后(0.79±0.31,P<0.05);在皮损部位表皮中为1.69±0.26,明显高于正常人对照组(0.89±0.37,P<0.01)与治疗后患者(O.91±0.32,P<0.01),在皮损部位真皮中为1.75±0.43,明显高于正常人对照组(0.79±0.28,P<0.001)及治疗后患者(0.98±0.26,P<0.01)。结论CCR2可能主要通过参与活化、趋化淋巴细胞而在银屑病的发病机制及病理状态的维持中起到重要作用,NB-UVB对银屑病的治疗作用可能通过调节CCR2的表达而实现。     

斑块状银屑病表皮干扰素-γ受体mRNA表达的研究

目的 探讨斑块状银屑病患者表皮干扰素-γ受体mRNA的表达及其在发病机制中的作用.方法 采集28例斑块状银屑病患者皮损及周围外观正常皮肤,28例正常人皮肤作为对照,分离表皮,逆转录聚合酶链反应(RT-PCR)法检测干扰素-γ受体mRNA表达水平.PASI评分法评估银屑病的严重程度.结果 斑块状银屑病患者皮损和非皮损表皮干扰素-γ受体mRNA的表达率为100%,对照样本的表达率为17.86%(5/28);患者皮损、非皮损和对照表皮干扰素-γ受体mRNA的表达水平均值分别为1.002±0.563、0.188±0.095、0.005±0.012,皮损处的表达水平明显高于非皮损处(t=7.54,P<0.01),非皮损处明显高于正常人对照;进行期与稳定期皮损的表达水平分别为1.210±0.489和0.379±0.163,进行期显著高于稳定期(t=4.37,P<0.01);而皮损处的表达水平与PASI评分值无相关性.结论 表皮干扰素-γ受体mRNA的表达可能与银屑病皮损的形成和病情的活动性有关.     

寻常型银屑病患者外周血环氧合酶同工酶的研究

目的 探讨进行期斑块状银屑病患者外周血单一核细胞（PBMC）中环氧合酶同工酶的表达及其在银屑病发病机制中的作用。方法 应用逆转录－聚合酶链反应（RT－PCR）检测了进行期斑块状银屑病患者及健康对照者各30例PBMC中构成性环氧合酶（COX－1）和诱导性环氧合酶（COX－2）mRNA的表达。结果 进行期斑块状银屑病患者PBMC中COX－2 mRNA的平均表达水平明显高于健康对照组（P＜0．01），且其mRNA的表达水平与其相应的面积与严重度积分（PASI）呈正相关（P＜0．01）；而两组间COX－1平均表达水平的差异无显著性（P＞0．05）。结论 进行期斑块状银屑病患者PBMC中COX－2 mRNA的表达增高，提示可能通过诱导前列腺素（PGs）的产生参与进行期斑块状银屑病的发病。     

寻常性银屑病患者外周血中性粒细胞中趋化因子受体的表达

目的:了解趋化因子受体在寻常性银屑病外周血中性粒细胞活化及趋化中的作用,探讨寻常性银屑病的发病机制。方法:应用反转录(RT)-PCR检测33例进行期斑块状银屑病患者及其中16例患者治疗后外周血中性粒细胞中趋化因子受体CXCR1～3及CCR1、5mRNA的表达水平,并设30名正常人作对照。将检测结果与银屑病皮损面积严重度指数(PASI)进行相关性分析。结果:寻常性银屑病患者治疗前中性粒细胞中CXCR1、CXCR2与CCR1mRNA表达水平分别为1.36±1.19、1.45±0.87与1.43±1.28,明显高于正常对照及治疗后的患者[正常对照分别为0.55±0.37(P<0.01),0.74±0.58(P<0.001),0.68±0.37(P<0.001);治疗后分别为0.46±0.33(P<0.001),0.63±0.58(P<0.01),0.73±0.86(P<0.05)],其中CXCR1、CXCR2与PASI呈显著正相关。结论:CXCR1、CXCR2与CCR1在寻常性银屑病患者外周血中性粒细胞中表达异常增高,它们可能参与了中性粒细胞的活化及趋化,进而参与了银屑病的发病。     

系统性红斑狼疮患者外周血单一核细胞CXCR3、CXCR5 mRNA的表达

目的: 检测趋化因子受体CXCR3及CXCR5在系统性红斑狼疮(SLE)患者外周血单一核细胞(PBMC)中的表达,并分析其与疾病活动性的相关性.方法: 收集93例SLE患者和30例正常对照者的外周血单一核细胞,提取RNA.应用RT-PCR方法检测CXCR3、CXCR5 mRNA的表达.结果: 活动期SLE CXCR3 mRNA的表达高于健康对照组.活动期SLE患者CXCR5 mRNA的表达均高与非活动期和健康对照组.SLE患者组CXCR5 mRNA表达水平与SLE疾病活动指数(SLEDAI)呈正相关.结论:CXCR3和CXCR5 mRNA的高表达提示SLE处于活动期.     

黄芪注射液对寻常性银屑病患者外周血单核细胞白介素-2mRNA表达的影响

目的观察黄芪注射液对寻常性银屑病患者外周血单核细胞白介素-2mRNA(IL-2mRNA)表达的影响,探讨黄芪注射液治疗银屑病的可能机制。方法随机抽取20例寻常性银屑病患者和20例正常对照者的外周血,采用逆转录聚合酶链反应(RT-PCR)法检测IL-2mRNA的表达。将20例寻常性银屑病患者外周血单核细胞与不同浓度的黄芪注射液共培养后,用RT-PCR法检测其IL-2mRNA的表达水平。结果寻常性银屑病患者外周血IL-2mRNA表达水平(1.230±0.052)明显高于正常对照组(0.807±0.075),差异有统计学意义(P<0.05);低浓度组IL-2mRNA表达水平(1.214±0.060)与空白对照组(1.230±0.052)差异不显著(P>0.05);中浓度组IL-2mRNA表达水平(1.393±0.066)明显高于空白对照组,高浓度组IL-2mRNA表达水平(1.113±0.041)明显低于空白对照组,差异均有统计学意义(P均<0.05)。结论低浓度黄芪注射液对IL-2mRNA的表达水平无影响,中浓度黄芪注射液可促进IL-2mRNA表达,高浓度黄芪注射液抑制IL-2mRNA的表达,黄芪注射液治疗寻常性银屑病可能通过影响IL-2mRNA的表达来发挥作用。     

MiR-146a在寻常型银屑病患者外周血单个核细胞中的表达及其临床意义

目的:检测寻常型银屑病患者外周血单个核细胞(PBMC)中miR-146a的表达情况,并探讨其与寻常型银屑病发病的相关性。方法:应用实时荧光定量聚合酶链反应检测30例寻常型银屑病患者外周血PBMC中的miR-146a的相对表达量,与30例正常对照组进行比较并对其与寻常型银屑病患者PASI评分行相关性分析。结果:寻常型银屑病患者外周血PBMC中的miR-146a的表达明显高于正常对照组,进行期患者组明显高于静止期患者组,差异均有统计学意义(P值均<0.05)。寻常型银屑病患者外周血PBMC中miR-146a的表达与PASI评分呈正相关性(r=0.62,P<0.05)。结论:寻常型银屑病患者外周血PBMC中miR-146a异常高表达,并与寻常型银屑病的分期相关,提示miR-146a在寻常型银屑病的发病中起重要作用。     

阿维A对中、重度斑块状银屑病患者Th17细胞相关因子的影响

目的探讨阿维A治疗中、重度斑块状银屑病的疗效以及对银屑病患者血清Th17细胞相关因子IL-17,IL-22及IL-23的影响,进一步阐明阿维A治疗寻常性银屑病的作用机制。方法 35例中、重度斑块状银屑病患者口服阿维A治疗8周,以银屑病皮损面积和严重程度(PASI)评分评价疗效;采用ELISA法检测健康对照组以及银屑病治疗组使用阿维A治疗前后血清IL-17,IL-22及IL-23的水平。结果阿维A治疗前后PASI评分明显下降(P<0.01);银屑病组和健康对照组比较,IL-17,IL-22和IL-23的血清水平均明显增高(P<0.01);阿维A治疗后血清中IL-17的水平(45.38±11.69)pg/mL,IL-22的水平(61.48±18.76)pg/mL及IL-23的水平(139.65±40.28 pg/mL),较治疗前显著下降(P均<0.01)。结论银屑病患者的血清中存在高水平的IL-17,IL-22和IL-23;阿维A治疗中、重度银屑病疗效明显,治疗后血清中IL-17,IL-22和IL-23明显降低;阿维A可能通过影响Th17细胞相关因子来发挥治疗银屑病的作用。     

Autocrine regulation of re-epithelialization after wounding by chemokine receptors CCR1, CCR10, CXCR1, CXCR2, and CXCR3

This study identifies chemokine receptors involved in an autocrine regulation of re-epithelialization after skin tissue damage. We determined which receptors, from a panel of 13, are expressed in healthy human epidermis and which monospecific chemokine ligands, secreted by keratinocytes, were able to stimulate migration and proliferation. A reconstructed epidermis cryo(freeze)-wound model was used to assess chemokine secretion after wounding and the effect of pertussis toxin (chemokine receptor blocker) on re-epithelialization and differentiation. Chemokine receptors CCR1, CCR3, CCR4, CCR6, CCR10, CXCR1, CXCR2, CXCR3, and CXCR4 were expressed in epidermis. No expression of CCR2, CCR5, CCR7, and CCR8 was observed by either immunostaining or flow cytometry. Five chemokine receptors (CCR1, CCR10, CXCR1, CXCR2, and CXCR3) were identified, the corresponding monospecific ligands (CCL14, CCL27, CXCL8, CXCL1, CXCL10, respectively) of which were not only able to stimulate keratinocyte migration and/or proliferation but were also secreted by keratinocytes after introducing cryo-wounds into epidermal equivalents. Blocking of receptor-ligand interactions with pertussis toxin delayed re-epithelialization, but did not influence differentiation (as assessed by formation of basal layer, spinous layer, granular layer, and stratum corneum) after cryo-wounding. Taken together, these results confirm that an autocrine positive-feedback loop of epithelialization exists in order to stimulate wound closure after skin injury.     

Delayed wound healing in CXCR2 knockout mice

Previous studies demonstrated that the CXC chemokine, MGSA/GRO-alpha and its receptor, CXCR2, are expressed during wound healing by keratinocytes and endothelial cells at areas where epithelialization and neovascularization occur. The process of wound healing is dependent on leukocyte recruitment, keratinocyte proliferation and migration, and angiogenesis. These processes may be mediated in part by CXC chemokines, such as interleukin-8 and MGSA/GRO-alpha. To examine further the significance of CXC chemokines in wound healing, full excisional wounds were created on CXCR2 wild-type (+/+), heterozygous (+/-), or knockout (-/-) mice. Wounds were histologically analyzed for neutrophil and monocyte infiltration, neovascularization and epithelialization at days 3, 5, 7, and 10 postwounding. The CXCR2 -/- mice exhibited defective neutrophil recruitment, an altered temporal pattern of monocyte recruitment, and altered secretion of interleukin-1beta. Significant delays in wound healing parameters, including epithelialization and decreased neovascularization, were also observed in CXCR2 -/- mice. In vitro wounding experiments with cultures of keratinocytes established from -/- and +/+ mice revealed a retardation in wound closure in CXCR2 -/- keratinocytes, suggesting a role for this receptor on keratinocytes in epithelial resurfacing that is independent of neutrophil recruitment. These in vitro and in vivo studies further establish a pathophysiologic role for CXCR2 during cutaneous wound repair.     

Human T lymphocytes and mast cells differentially express and regulate extra- and intracellular CXCR1 and CXCR2

CXCL8 plays a major role in cell recruitment to sites of inflammation. Apart from neutrophils, little is known, however, about the cellular distribution and regulation of CXCL8 receptors in cells involved in acquired and adaptive immune responses. In previous studies, we have demonstrated the extracellular expression and function of CXCR1/2 on mast cells and also detected an intracellular pool of CXCR1/2. Here, we have addressed the question of receptor regulation during stimulation of human mast cells (HMC-1 cell line) and have studied T cells in comparison. Cell permeabilization was performed to detect both surface and possible intracellular receptor pools. HMC-1 cells stained positive for both receptors on the cell surface (CXCR1, 50%; CXCR2, 51%) and also after cell permeabilization (CXCR1, 86%; CXCR2, 74%). Similarly, T cells exhibited both cell-surface receptor expression (CXCR1, 30%; CXCR2, 23%) and higher total receptor expression (CXCR1, 50%; CXCR2, 36%), although overall values were lower than that in HMC-1 cells. On immunoblot, molecular weights of extra- and intracellular receptors on mast cells were the same, excluding altered receptor glycosylation. On stimulation with phorbol 12-myristate 13-acetate plus calcium ionophore, a time-dependent decrease of surface-membrane receptors was observed in both cell types, while total receptor remained the same, suggesting that receptor shedding is not involved. The kinetics of membrane receptor internalization and replenishment differed for the two cell types. Furthermore, receptor internalization was associated with decreased F-actin polymerization, a basic prerequisite for cell migration. These findings demonstrate for the first time the expression of extra- and intracellular CXCR1/2 receptors on T cells and delineate the dynamics of CXCR1/2 receptors on mast cells and T cells. Furthermore, they suggest a cell-type-specific and finely tuned regulation of chemokine responses at the receptor level in the context of inflammation.