皮肤肿瘤中Wnt/β-连环蛋白信号通路的研究进展

Wnt/β-连环蛋白信号通路的异常活化常与皮肤肿瘤的发生发展密切相关.导致通路异常的因素较多,如分泌性Wnt蛋白失调;通路蛋白分子或旁路调控分子的突变;病毒感染和其他通路交叉影响等.通路的致瘤机制及对肿瘤的生物学影响还存在组织特异性,而皮肤肿瘤种类多,且组织来源广泛.因此探索Wnt通路在皮肤肿瘤中差异及组织特异性将对肿瘤靶向治疗研究有指导意义.

Abstract：

The abnormal activation of Wnt/β-catenin signaling pathway is usually and closely related to the occurrence and development of cutaneous tumors.There are various factors causing the abnormality of this pathway, such as deregulated expression of secreted Wnt protein, mutations of proteins involved in this pathway or regulatory molecules in bypass pathway, viral infection, interaction with other pathways, and so on.In addition, the tumorigenic mechanisms of this pathway and its biologcal effects on tumors are considered to be tissue-specific.However, the types and tissue origins of cutaneous tumors are various.Therefore, exploring the disparities and tissue specificity of Wnt pathway in cutaneous tumors will provide a guide to the targeted therapy of tumors.     

β联蛋白对体外过氧化氢所致人皮肤成纤维细胞衰老表型的影响

目的 观察高表达的β联蛋白对体外过氧化氢（H2O2）所致正常人皮肤成纤维细胞（NHSF）衰老表型的影响。方法 NHSF分别转染pcDNA3.1-β联蛋白或空载体pcDNA3.1,然后150 μmol/L H2O2处理2 h。实验分为转染H2O2处理组（NHSF + β联蛋白 + H2O2）、转空载体H2O2处理组（NHSF + 空载体 + H2O2）以及转空载体组（NHSF + 空载体）。用RT-PCR和Western印迹分析β联蛋白的表达,显微镜观察细胞形态,试剂盒检测衰老相关的β半乳糖苷酶（SA-β-gal）、细胞活性氧（ROS）以及超氧化物歧化酶（SOD）的活性。所有数据均用SPSS 13.0软件进行统计分析,多组间比较采用ANOVA检验。结果 pcDNA3.1-β联蛋白明显上调了NHSF中β联蛋白的表达。NHSF + 空载体 + H2O2组β联蛋白mRNA和蛋白水平（与GAPDH mRNA和蛋白的比值分别为0.2900 ± 0.0195和0.3130 ± 0.0171）明显低于NHSF + 空载体组（分别为0.5963 ± 0.0400和0.6190 ± 0.0090）,两组比较,均P ＜ 0.05;NHSF + β联蛋白 + H2O2组β联蛋白表达水平（0.7953 ± 0.0074）高于NHSF + 空载体 + H2O2组（P ＜ 0.05）。SA-β-gal染色率在NHSF + 空载体组、NHSF + 空载体 + H2O2组、NHSF + β联蛋白 + H2O2组分别为（2.9667 ± 0.2517）%、（37.70 ± 0.9539）%、（29.330 ± 0.6359）%,各组比较,差异有统计学意义（P ＜ 0.05）。NHSF + 空载体、NHSF + 空载体 + H2O2、NHSF + β联蛋白 + H2O2组ROS活性分别为（50.9963 ± 9.2688）%、（109.9190 ± 11.5215）%、（75.1063 ± 3.0138）%,SOD水平分别为（88.0856 ± 3.9181）、（35.5585 ± 3.4438）、（61.7029 ± 3.1716） U/mg,各组比较,差异均有统计学意义（P ＜ 0.05）。结论 高表达的β联蛋白能够抑制细胞衰老相关β-半乳糖甘酶（SA-β-gal）和细胞活性氧（ROS）的表达,同时上调了SOD的活性。 【关键词】 β连环素; 过氧化氢; 成纤维细胞; 氧化性应激; 细胞衰老     

Role of β2-microglobulin in uremic patients may be greater than originally suspected

The role of beta2-microglobulin(β2M) in dialysisrelated amyloidosis as a specific amyloid precursor was defined in the 1980 s. Studies in those years were largely related to β2M amyloidosis. In 2005, for what was probably the first time in the available literature, we provided data about the association betweenβ2M and early-onset atherosclerosis in hemodialysis patients without co-morbidities. In recent years, the role of uremic toxins in uremic atherosclerosis and the interest in β2M as a marker of cardiovascular(CV) and/or mortality risk have grown. In the current literature,clinical studies suggest that β2M is an independent, significant predictor of mortality, not only in dialysis patients, but also in predialysis patients and in the highrisk portion of the general population, and it seems to be a factor strongly linked to the presence and severity of CV disease. It is still unknown whether β2M is only a uremic toxin marker or if it also has an active role in vascular damage, but data support that it may reflect an increased burden of systemic atherosclerosis in a setting of underlying chronic kidney disease. Thus, although there have been some inconsistencies among the various analyses relating to β2M, it promises to be a novel risk marker of kidney function in the awareness and detection of high-risk patients. However, more research is required to establish the pathophysiological relationships between retained uremic toxins and further biochemical modifications in the uremic milieu to get answers to the questions of why and how. In this review, the recent literature about the changing role of β2M in uremic patients will be examined.     

β-连环蛋白在毛母质瘤及毛发上皮瘤中的表达

目的: 检测β-连环蛋白(β-catenin)在毛母质瘤及毛发上皮瘤中的表达.方法: 应用免疫组织化学法检测20例正常皮肤、20例毛母质瘤及10例毛发上皮瘤中β-连环蛋白的表达.结果: β-连环蛋白在正常皮肤角质形成细胞中主要表达于细胞膜, 15例毛母质瘤和8例毛发上皮瘤的表达位于胞质和/或胞核.结论: β-连环蛋白可能参与毛基质细胞的增殖和成熟的调控,它的异常表达在毛囊发育和向毛囊分化肿瘤的发生中可能起重要作用.     

黄芪甲苷对光老化小鼠皮肤中转化生长因子-β信号通路的影响

目的 探讨黄芪甲苷对皮肤光老化的保护机制.方法 BALB/c小鼠分为模型组、模型+基质组、模型+黄芪甲苷组、正常对照组.RT-PCR测定转化生长因子β受体Ⅱ(TGF-βRⅡ)、Smad 7的mRNA表达水平,免疫组化观察TGF-βR Ⅱ、Smad 7在小鼠皮肤组织中的蛋白表达情况.结果 正常对照组中,TGF-βR Ⅱ、Smad 7的灰度值比值分别为0.5688±0.0439、0.5900 ±0.0585,阳性表达率分别为(53.00 ±4.72)%、(47.50±3.81)%;模型组中,TGF-βR Ⅱ、Smad 7的灰度值比值分别为0.2588±0.0283、0.8637±0.0514,阳性表达率分别为(28.20±5.24)%、(82.06±2.18)%;模型+基质组中,TGF-βR Ⅱ、Smad 7的灰度值比值分别为0.2653±0.0456、0.8553±0.0575,阳性表达率分别为(28.74±2.28)%、(82.62±4.02)%;模型+黄芪甲苷组中TGF-βRⅡ、Smad 7的灰度值比值分别为0.3767±0.0374、0.7131±0.0410,阳性表达率分别为:(41.64±2.59)%、(64.36±2.62)%.皮肤组织中TGF-βRⅡ和Smad灰度值比值4组小鼠间比较,F值分别为80.98和736.80,TGF-βRⅡ和Smad 7的阳性表达率4组小鼠间比较,F值分别为45.36和132.25,P值均<0.01.与正常对照组相比,模型组TGF-βR Ⅱ mRNA和蛋白表达明显降低,Smad 7 mRNA和蛋白表达显著升高(P值均<0.01).与模型组和模型+基质组比较,模型+黄芪甲苷组TGF-βRⅡ mRNA和蛋白表达明显升高,Smad 7 mRNA和蛋白表达显著下调(P值均<0.01).模型+基质组TGF-βR Ⅱ、Smad 7的mRNA和蛋白表达与模型组相比差异无统计学意义(P值均>0.01).结论 黄芪甲苷可以通过上调TGF-βRⅡ表达和下调Smad 7表达而改变TGF-β通路的信号转导参与抗光老化.

Abstract：

Objective To study the protective mechanism of astragaloside on skin photoaging. Methods BALB/c mice were randomly divided into four groups: model group irradiated with ultraviolet rays (UV), model plus matrix group pretreated with the matrix before UV irradiation, model plus astragaloside group pretreated with astragaloside 0.08% cream before UV irradiation, normal control group received no irradiation or pretreatment. After 4-week irradiation, the mice were sacrificed, and skin tissues were resected from the back of these mice. Then, reverse transeription PCR (RT-PCR) and immunohistochemistry were performed to detect the mRNA and protein expression of TGF-βR Ⅱ and Smad 7, respectively. Gray scale ratio was used to represent the mRNA levels of TGF-βR Ⅱ and Smad 7. Results There was a significant difference in the mRNA level (F = 80.98, 736.80, respectively, both P < 0.01) and protein positivity rate (F = 45.36,132.25, respectively,both P < 0.01) of TGF-βR Ⅱ and Smad 7 among the 4 groups. The mRNA level and protein positivity rate of TGF-βR Ⅱwere 0.2588±0.0283 and (28.20 ± 5.24)% respectively in the model group, significantly lower than those in the normal control group[0.5688 ± 0.0439, (53.00 ± 4.72)%, both P < 0.01] and model plus astragaloside group [0.3767 ± 0.0374, (41.64 ± 2.59)%, both P< 0.01]; on the contrary, the mRNA level and protein positivity rate of Smad 7 in the model group [0.8637 ± 0.0514, (82.06 ± 2.18)%] were significantly higher than those in the normal control group [0.5900 ± 0.0585, (47.50±3.81)%, both P < 0.01] and model plus astragaloside group [0.7131 ± 0.0410, (64.36 ± 2.62)%, both P< 0.01]. In the model plus astragaloside group, the mRNA level and protein positivity rate of TGF-βR Ⅱ were significantly higher than in the model plus matrix group [0.2653 ± 0.0456, (28.74 ± 2.28)%, both P < 0.01], while those of Smad 7 were statistically lower than in the model plus matrix group [0.8553 ± 0.0575, (82.62 ± 4.02)%, both P < 0.01]. However,no significant difference was observed in the mRNA level or protein positivity rate of TGF-βR Ⅱ or Smad 7 between the model group and model plus matrix group (all P > 0.01). Conclusion Astragaloside can prevent skin photoaging by the alteration of TGF-β pathway via up-regulating TGF-βR Ⅱ expression and down-regulating Smad 7 expression.     

H2O2对人皮肤成纤维细胞衰老标记蛋白30及自噬相关蛋白LC3Ⅱ表达的影响北大核心CSCD

目的 探讨H2O2对人皮肤成纤维细胞（NHSF）衰老标记蛋白30（SMP30）以及细胞自噬相关蛋白微管相关蛋白1轻链3Ⅱ型（LC3Ⅱ）的影响。方法 用150 μmol/L H2O2处理2 ~ 4代NHSF 2 h,构建细胞衰老模型作为H2O2组,而未处理的NHSF作为对照组。细胞衰老β半乳糖苷酶（SA？β？gal）染色法检测衰老细胞比例,间接免疫荧光检测自噬相关蛋白LC3的表达,反转录PCR（RT？PCR）检测SMP30 mRNA的表达,Western印迹法检测SMP30和LC3蛋白的表达。结果 H2O2组NHSF的SA？β？gal染色阳性率（41.70% ± 2.95%）显著高于对照组（3.03% ± 0.25%,t = 22.59,P 〈 0.05）。间接免疫荧光结果显示,H2O2组NHSF LC3阳性率（12.60% ± 1.57%）明显低于对照组（23.67% ± 3.04%,t = 5.61,P 〈 0.05）。Western印迹显示,H2O2组LC3Ⅰ/GAPDH比值（0.40 ± 0.02）与对照组（0.41 ± 0.04）比较差异无统计学意义（P 〉 0.05）,而H2O2组LC3Ⅱ/GAPDH比值（0.20 ± 0.02）明显低于对照组（0.80 ± 0.03,t = 29.69,P 〈 0.05）,且H2O2组LC3Ⅱ/LC3Ⅰ比值（0.51 ± 0.03）亦明显低于对照组（1.98 ± 0.23,t = 10.967,P 〈 0.05）。H2O2组SMP30 mRNA和蛋白水平（与GAPDH mRNA和蛋白的比值分别为0.16 ± 0.01和0.27 ± 0.02）明显低于对照组（分别为0.35 ± 0.01和0.63 ± 0.02）,均P 〈 0.05。结论 H2O2可降低NHSF中SMP30及LC3Ⅱ的表达水平,加速NHSF衰老。     

Human β-defensin-2 and skin diseases

Human β-defensin-2 (hBD-2) is an endogenous antimicrobial peptide recently found in the skin with broad-spectrum antimicrobial activity and unique mechanism of function. Recent studies have confirmed that the expression of hBD-2 is upregulated in the lesions of some dermatoses, such as psoriasis,acne vulgaris, verruca, dermatophytosis and basal cell carcinoma, etc. The susceptibility to bacterial and viral infections may be associated with the decreased or absent expression of hBD-2. Further researches into HBD-2will provide a new direction for the prevention and treatment of skin diseases.     

炎症浸润细胞在曝光和非曝光部位皮肤中的组织学表现

目的 观察、比较曝光及非曝光部位皮肤中炎症浸润细胞的类型、数目,探讨其在光老化过程中的作用.方法 应用免疫组化法分别对23例女性健康志愿者前臂伸侧(曝光)和上臂内侧(非曝光部位)皮肤石蜡标本中的细胞表面抗原CD3、CD45RO、CD68进行检测,计数阳性细胞数目.不同部位阳性细胞数比较采用配对t检验,阳性细胞数与年龄的相关性采用Pearson相关分析.结果 曝光部位皮肤组织CD3、CD45RO、CD68阳性细胞数分别为(48.91±13.17)/mm2、(46.83±12.92)/mm2、(85.43±22.35)/mm2,非曝光部位分别为(40.61±11.57)/mm2、(38.00±10.11)/mm2、(73.48±16.21)/mm2,曝光部位均显著高于非曝光部位(P<0.01或<0.05).曝光部位皮肤组织CD3、CD45RO阳性细胞数与年龄呈正相关(r=0.56、0.56,P<0.01),曝光部位CD68及非曝光部位皮肤组织CD3、CD45RO、CD68阳性细胞数与年龄无相关性.结论 T淋巴细胞、巨噬细胞可能在光老化过程中发挥作用.

Abstract：

Objective To investigate the role of infiltrating inflammatory cells in photoaging process by comparing the type and number of these cells in sun-exposed and-unexposed skin.Methods The expression of CD3,CD45RO and CD68 were detected by immunohistochemieal staining in 46 paraffin-embeded skin samples from the extensor forearms(sun-exposed)and upper-inner arms(sun-unexposed) of 23 healthy female volunteers.The number of positive cells in sun-exposed and -unexposed sites was counted and statistically tested by paired samples t test,and Pearson correlation analysis was performed to assess the relationship between the number of positive cells and age of these volunteers.Results The number of cells positive for CD3,CD45RO and CD68 per square millimetre in sun-exposed skin was significantly higher than that in sun-unexposed skin(48.91±13.173 vs.40.61±11.571,46.83±12.915 vs.38.00±10.109,85.43±22.346 vs.73.48±16.208,respectively,P<0.01 or 0.05).The number of cells positive for CD3 and CD45RO increased significantly with age (r=0.557,0.555,respectively,both P<0.01) in the sun-exposed skin but not in sun-unexposed skin,and the number of CD68-positive cells was uncorrelated with age in either sunexposed or -unexposed skin.Conclusion T lymphocytes and macrophages may play a role in the process of photoaging.     

人皮肤鳞状细胞癌A431细胞株电压门控钾通道的研究

目的 探讨电压门控钾通道在人皮肤鳞状细胞癌A431细胞和人角质形成细胞HaCaT细胞中的作用.方法 噻唑蓝方法研究加入含有不同浓度四乙胺的培养基对A431细胞和HaCaT细胞的增殖的影响;提取体外培养A431细胞、HaCaT细胞以及人正常表皮组织的蛋白,ELISA方法检测电压门控钾通道蛋白(HERG)在它们中的表达,比较它们的差异.结果 四乙胺对体外培养的A431细胞和HaCaT细胞的增殖抑制效应具有时间依赖与剂量依赖的特点,当四乙胺的浓度≥10 mmol/L且作用时间≥24 h,可以使A431细胞和HaCaT细胞的增殖受到明显的抑制.HERG钾通道蛋白在A431细胞、HaCaT细胞和人正常皮肤表皮组织均有表达,平均浓度分别为(49.7114±3.55696)pg/ml、(35.7471±4.14696)pg/ml、(36.8857±3.47810)pg/ml,组间比较差异具有统计学意义(P<0.05).结论 阻断电压门控钾通道可抑制A431细胞和HaCaT细胞的增殖;电压门控钾通道可能在人皮肤鳞状细胞癌细胞上的表达更显著.

Abstract：

Objective To investigate the role of voltage-gated potassium channel in the human skin squamous cell carcinoma A431 cells and human keratinocyte HaCaT cells. Methods MTT assay was performed to detect the effect of different concentrations of tetraethylammonium (TEA) on the proliferation of cultured A431 cells and HaCaT cells. Besides, enzyme-linked immunosorbent assay (ELISA) was conducted to detect the expression of HERG channel protein in A431 cells, HaCaT cells and normal human skin tissue.Results TEA inhibited the proliferation of A431 cells and HaCaT cells in a dose- and time-dependent manner.After treated with TEA (≥ 10 mmol/L) for 24 or more hours, the proliferation of A431 cells and HaCaT cells was obviously suppressed. A significant difference was observed in the average concentration of HERG channel protein between A431 cells, HaCaT cells and normal human skin tissue (49.7114 ± 3.55696 pg/ml, 35.7471 ±4.14696 pg/ml, 36.8857 ± 3.47810 pg/ml, all P < 0.05). Conclusions The block of voltage-gated potassium channel could inhibit the proliferation of A431 cells and HaCaT cells, and the expression of voltage-gated potassium channel seems to be higher in human skin squamous cell carcinoma cells.     

皮肤细胞DNA光损伤后反应机制研究进展

紫外线可引起皮肤细胞DNA损伤,诱发皮肤肿瘤和光老化.DNA损伤类型包括:DNA单链断裂、链间交联、核苷酸碱基修饰等.细胞拥有DNA损伤后反应机制以维持基因组稳定,如细胞周期监控点、DNA修复和细胞早衰等.细胞周期监控系统检测到染色体结构异常时将引起细胞周期停滞并启动修复进程;而在检测到无法修复的损伤时则会诱使细胞衰老.因此皮肤光老化也是一种预防肿瘤发生的防御机制,ATR-Chk1信号通路在其中起着关键的调控作用.

Abstract：

Ultraviolet radiation can induce DNA damage in skin cells, cause skin tumors and accelerate skin photoaging. UV-induced DNA damage in skin cells includes DNA single strand breaks, DNA interstrand cross-links and nucleotide base modifications. Skin cells could exert DNA damage responses, such as cell cycle checkpoints, DNA repair and premature senescence to prevent genomic instability. When cell cycle checkpoint systems sense the abnormal chromosomal DNA structures, they execute cell cycle arrest and DNA repair process will be initiated. Cellular senescence is also executed by checkpoint responses when unrepairble and extensive chromosomal abnormalities are detected. So skin photoaging is thought to be one of the major mechanisms against skin carcinogenesis. ATR-Chk1 signaling pathway plays a key role in the regulation of DNA damage response process.     

寻常性银屑病患者外周血T细胞受体β可变区优势表达分析

目的 分析寻常性银屑病患者外周血中T细胞受体β可变区(TRBV)的优势表达状况,探讨其在银屑病发病中的作用.方法 以人功能性TRBV家族设计33个上游引物,共同的T细胞受体β恒定区(TRBC)基因设计下游引物,在T细胞受体α恒定区(TRAC)设计引物作为内参,分析寻常性银屑病患者、正常人对照外周血样本各10例,检测各PCR反应管荧光强度,以大于正常人外周血T细胞相应TRBV基因相对表达量(-x+3s)筛选TRBV优势表达基因家族.结果 TRAC扩增产物阈值循环数(Ct)值集中于21～24,银屑病组TRBV2扩增产物Ct与TRAC产物Ct差值平均数银屑病患者为2.98,TRBV5-7为3.24,TRBV6-6/6-9为2.52,TRBV12为2.04,TRBV24为3.56,TRBV29为4.12,其表达与正常人比较均明显增高(P值均＜0.05),其中TRBV6-6/6-9、TRBV12、TRBV29在银屑病患者呈优势表达.结论 银屑病患者外周血TRBV基因家族存在优势表达,可能在银屑病T细胞异常免疫反应中起重要作用.

Abstract：

Objective To assess the preferential expressions of peripheral blood T cell receptor beta chain variable region (TRBV) subfamilies in patients with psoriasis vulgaris(PV), and to estimate their role in the pathogenesis of psoriasis. Methods Thirty-three upstream primers were designed to target the human functional TRBV genes, downstream primers to target the common T cell receptor beta constant (TRBC) gene,with T cell receptor alpha constant (TRAC) gene as the internal reference. Total RNA was extracted from the peripheral blood T cells of 10 health human controls and 10 patients with PV, and transcribed into cDNA.Then, TRBV genes were amplified by real-time fluorescence quantitative PCR (RFQ-PCR) and the fluorescence intensity of each samples was detected. The expression levels of TRBV genes in the control group were used to calculate the cut-off values (mean expression levels of TRBV subfamilies in the 10 normal controls + 3 standard deviations). When the expression level of a TRBV subfamily from patients with PV was equal to or higher than the cut-off value, it was considered as the preferentially expressed TRBV subfamily. Results The threshold cycle (Ct) value varied from 21 to 24 for TRAC gene. The difference in the Ct value between TRBV subfamily genes and TRAC gene in patients with PV was 2.98 for TRBV2 gene, 3.24 for TRBV5-7 gene, 2.52 for TRBV6-6/6-9 gene, 2.04 for TRBV 12 gene, 3.56 for TRBV 24 gene, and 4.12 for TRBV 29 gene, and the expression levels of these subfamily genes were significantly higher than those in the normal controls (all P ＜ 0.05). According to the above standard, TRBV6-6/6-9, TRBV12 and TRBV29 were considered to be preferentially expressed subfamilies. Conclusions There is a preferential expression of TRBV gene subfamilies in peripheral blood of patients with psoriasis vulgaris, which may play a vital role in the abnormal T cell-mediated immune responses in psoriasis.     

Alleviating Skin Barrier Disruption,Skin Inflammation,and Pruritus: A Moisturizing Spray Containing β-Glucan and Panthenol

Objective: Inflammatory skin diseases were proved to be associated with dry skin-induced pruritus. However, the relationship between skin inflammation, skin barrier function, and pruritus remains unclarified. The present study aimed to explore this relationship using an acetone-ether-water (AEW) mouse model, and to investigate the anti-itch effects of the combined application of β-glucan and panthenol in a moisturizing spray in this mouse model.Methods: A dry skin-induced chronic pruritus mouse ...     

磷酸化NF-κB亚基P65介导化学性缺氧诱导的HaCaT细胞炎症损伤

目的 探讨核因子-κB(NF-κB)亚基P65磷酸化是否参与化学性缺氧模拟剂氯化钴(CoCl2)诱导的永生化人皮肤角质形成细胞(HaCaT)毒性作用及炎症反应.方法 用2 mmol/L CoCl2处理HaCaT细胞,建立化学性缺氧诱导HaCaT细胞损伤的体外模型.RNA干扰法下调NF-κB亚基P65的表达.细胞计数试剂盒-8比色法检测细胞存活率;ELISA法检测培养基中IL-6和IL-8的含量;Western印迹法检测总量P65及磷酸化P65的蛋白表达.结果 CoCl2处理HaCaT细胞0～4 h,可促进NF-κB亚基P65的磷酸化,在0.5 h时P65亚基开始磷酸化,1.5 h时P65亚基的磷酸化水平达到高峰,约为对照组的6.6倍,而4 h基本恢复到正常水平.CoCl2处理HaCaT细胞0～6 h,可时间依赖性地降低细胞活力,2、4和6 h的细胞存活率与对照组比较,P值分别＜0.05、＜0.01及＜0.01.CoCl2处理6 h,还引起IL-6和IL-8的释放显著增加.RNA干扰法下调P65的表达后,CoCl2处理引起的HaCaT细胞毒性作用被明显减弱,即使细胞存活率升高了11%左右,下调P65表达还明显抑制了CoCl2处理引起的IL-6和Il-8释放增多.结论 磷酸化NF-κB亚基P65介导CoCl2诱导的HaCaT细胞毒性及炎症反应.

Abstract：

Objective To explore whether the phosphorylation of NF-κB P65 subunit is involved in the cytotoxicity to and inflammation in an immortal human keratinocyte cell line HaCaT during cobalt chloride (CoCl2-induced chemical hypoxia. Methods HaCaT cells were treated with CoCl2 of 2 mmol/L to set up a chemical hypoxia-induced cell model of injury. Then, RNA interference was used to down-regulate the expression of P65 in CoCl2-induced HaCaT cells. After additional culture, cell viability was tested by cell counting kit8 (CCK-8), the levels of interleukin 6 (IL-6) and interleukin 8 (IL-8) were detected by ELISA kits, phosphorylated and total P65 protein was measured by Western blot. Results The exposure of HaCaT cells to 2 mmol/L CoCl2 for 0 to 4 hours enhanced the phosphorylation of P65, which began at 0.5 hour, peaked at 1.5 hours, and restored to the normal level at 4 hours, and the level of P65 phosphorylation was about 6.6 times that in the untreated control group. The CoCl2 of 2 mmol/L decreased the cell viability of HaCaT cells in a time dependent manner, and a significant difference was observed in the viability of HaCaT cells between CoCl2-treated and untreated HaCaT cells at 2, 4, and 6 hours (P ＜ 0.05, 0.01, 0.01 ). The release of IL-6 and IL-8 from HaCaT cells was also promoted by CoCl2 treatment. The knockdown of P65 expression with siRNA markedly suppressed the CoCl2-induced cytotoxicity to and increase in the release of IL-6 and IL-8 from HaCaT cells,despite of an increment in cell viability by about 11%. Conclusion The phosphorylated P65 subunit mediates CoCl2-induced cytotoxicity and inflammatory injury to HaCaT cells.     

银屑病性关节炎患者血清中转化生长因子-β1的检测

目的 探讨银屑病性关节炎患者血清中转化生长因子(TGF)-β1的表达及其意义.方法 分别检测银屑病性关节炎患者与寻常性银屑病患者血清中TGF-β1的水平.结果 38例银屑病性关节炎患者血清中的TGF-β1浓度为375.92±95.88 pg/ml,80例寻常性银屑病患者为510.78±167.97 pg/ml,两组差异有统计学意义(t=4.60,P<0.01).结论 对银屑病性关节炎患者进行血清TGF-β1浓度的动态监测,有助于引起临床医师对银屑病性关节炎骨质疏松的警惕和预防.

Abstract：

Objective To measure the expression of TGF-β1 in the sera of patients with psoriatic arthritis and its significance.Methods The serum level of TGF-β1 was measured in 38 patients with psoriatic arthritis and 80 patients with psoriasis vulgaris.Results A significant difierence was observed between the patients with psoriatic arthritis and those with psoriasis vulgaris(375.92±95.883 pg/ml vs.510.78±167.967 pg/ml,t=4.60,P<0.01).Conclusion Continuous monitoring of serum levels of TGF-β1 may help clinicians to take precautions against osteoporosis in patients with psoriatic arthritis.     

树突细胞相关C型凝集素-1受体介导的抗白念珠菌免疫作用及机制

树突细胞相关c型凝集素-1是一种主要的C型凝集素样受体,存在于多种细胞和组织中.它能够特异性识别白念珠菌细胞壁主要成分β-葡聚糖,进而诱导机体产生固有免疫反应和适应性免疫反应.其可单独或与其他模式识别受体相互协同完成上述过程.白念珠菌β-葡聚糖的暴露、树突细胞相关C型凝集素-1表达水平等因素可影响机体免疫效应的强度,对其进行调控将为自念珠菌感染预防和治疗提供一个新的方向.

Abstract：

Dectin-1, a principal C-type lectin pattern-recognition receptor, exists in various types of cells and tissues. It can specifically recognize β-glucans, which are the major cell wall component of Candida albicans, and then trigger the innate and adaptive immune responses in hosts. Dectin-1 can independently, as well as collaboratively with other pattern recognition receptors, induce above mentioned processes. The intensity of immune responses in hosts is modulated by multiple factors such as the exposure of β-glucans on Candida albicans and the expression level of Dectin-1, and this modulation may provide a new direction for the prevention and therapy of Candida albicans infection.     

C型凝集素1受体参与人中性粒细胞体外杀伤白念珠菌活性的实验研究

【摘要】 目的 探讨人中性粒细胞能否通过C型凝集素1受体（dectin-1）识别白念珠菌细胞壁不溶性β葡聚糖来介导体外杀伤白念珠菌的活性。 方法 100 mg/L β葡聚糖体外作用于中性粒细胞1、6、24 h后,实时荧光定量逆转录PCR分析dectin-1和Toll样受体2 mRNA表达水平。100 mg/L β葡聚糖体外分别刺激中性粒细胞15 min、2 h、6 h,或先用dectin-1抑制剂昆布多糖100 mg/L和50 mg/L预处理30 min后,再予100 mg/L β葡聚糖刺激2 h,微量培养板荧光分析法检测中性粒细胞H2O2释放水平。昆布多糖预处理的中性粒细胞与白念珠菌体外共培养后,检测菌落形成单位（CFU）。结果 β葡聚糖作用中性粒细胞1、6、24 h后,dectin-1 mRNA水平分别为2.8195 ± 0.1669、5.4859 ± 0.7244、3.6041 ± 0.5372,均明显高于空白对照组（均P < 0.01）。β葡聚糖刺激中性粒细胞15 min后H2O2水平为（64.55 ± 15.67） μmol/L,2 h时为（290.34 ± 30.56） μmol/L,6 h时为（208.54 ± 26.88） μmol/L,与空白对照组（22.05 ± 3.99） μmol/L比较,差异均有统计学意义（均P < 0.01）;100 mg/L和50 mg/L昆布多糖预处理组分别为（80.45 ± 22.41） μmol/L和（130.42 ± 44.55） μmol/L,与β葡聚糖刺激组相比,分别降低了73%和45%,差异均有统计学意义（P < 0.01）。昆布多糖能明显抑制中性粒细胞体外杀伤白念珠菌的活性（均P < 0.01）。 结论 Dectin-1参与人中性粒细胞分泌H2O2以及对白念珠菌杀灭活性,为过继性粒细胞转输治疗系统性念珠菌感染提供初步依据。 【关键词】 念珠菌,白色; 中性白细胞; β葡聚糖类; 外源凝集素类,C型; 过氧化氢     

内皮素对黑素瘤A375细胞转化生长因子-β1及磷酸化Smad 3蛋白表达的调节

Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.