In situ expression of messenger RNA of interleukin-1 and interleukin-6 in psoriasis: interleukin-6 involved in formation of psoriatic lesions

Summary Psoriasis is a disease of abnormal proliferation and differentiation of epidermal cells. Several cytokines released by keratinocytes are implicated as factors responsible for this pathological condition of the epidermis. In order to elucidate the role of these cytokines in psoriasis, messenger RNA (mRNA) expression of interleukin-1 (IL-1) and IL-6 in psoriatic epidermis was investigated using biotin-labelled complementary DNA (cDNA) of the cytokines. Messenger RNA of IL-1 was weakly detected in some normal healthy epidermis specimens and more strongly in all the perilesional uninvolved psoriatic epidermis specimens. It was also expressed in the transitional zone between uninvolved and fully developed psoriatic skin, but was not expressed in lesional skin. In contrast, IL-6 mRNA was rarely expressed in normal healthy epidermis, but was expressed in perilesional uninvolved psoriatic epidermis, in the transitional zone and in the fully developed lesional epidermis, with the maximum intensity in the transitional zone. Expression of mRNA of IL-6 receptor showed a similar tendency to that of IL-6. It was expressed in psoriatic epidermis, most strongly in the transitional zone, but not in normal healthy epidermis. IL-6 was demonstrated immunohistochemically in psoriatic epidermis, but IL-6 receptor was demonstrated only in the transitional zone. Thus IL-6 and its receptor expression correlated well with the formation of psoriatic lesions where IL-1 may initiate their expression. IL-6 may play an important role in the pathogenesis of psoriasis.     

银屑病易感基因位点PSORS1的研究进展

银屑病是一种常见的多基因遗传性皮肤病,通过对不同群体的银屑病患者的基因样本进行全基因组扫描,确定了7个银屑病易感基因位点(PSORS1～PSORSS7),其中位于6p21染色体上的易感基因位点PSORS1是当前银屑病易感基因研究的热点,它包括三个银屑病候选基因:HLA-Cw6,CDSN,HCR.分别对这三个易感基因进行综述.     

ICAM-1 and LFA-1 Expressions in the Lesional Skin of Annular Erythema Associated with Sjögren Syndrome

ICAM-1 and LFA-1 expression was studied in the lesional skin of ten cases of annular erythema associated with Sjögren syndrome. Most of the infiltrating mononuclear cells around blood vessels expressed LFA-1 in addition to its strong expression on vascular endothelial cells and focal expression on the epidermal basal cell layer in 3 cases. ICAM-1 expression on vascular endothelial cells was similar to LFA-1, although relatively focal and weak expression was observed on mononuclear cells. ICAM-1 expression on keratinocytes was focal and limited to the basal cell layer in annular erythema. These findings suggest that strong expression of ICAM-1 on endothelial cells but not keratinocytes and LFA-1 on mononuclear cells might play some role in the induction of skin lesions in annular erythema associated with Sjögren syndrome.     

消异方对EM患者子宫内膜中MCP-1、IL-6免疫组化表达的影响

目的:探讨消异方对子宫内膜异位症( EMs)患者在位子宫内膜组织中MCP -1、IL -6免疫组化表达的影响.方法:将EMs患者90例随机分为空白对照组(30例)、中药治疗组(30例)、安慰剂对照组(30例),比较3组治疗前后在位子宫内膜中MCP -1、IL-6免疫组化的表达.结果:中药治疗组治疗前后在位子宫内膜中MCP -1、IL -6含量均较用药前明显降低,差别有统计学意义(P1＜0.05,P2＜0.05)；空白对照组和安慰剂对照组治疗前后在位子宫内膜中MCP -1、IL -6变化无统计学意义.结论:中药治疗能降低EMs患者子宫内膜中MCP -1、IL -6免疫组化表达.     

白介素1α及白介素6对角朊细胞与真皮成纤维细胞相互作用的影响

为了探讨细胞因子在表皮与真皮相互作用中所起的作用,用体外细胞培养的方法研究了白介素1α(IL-1α)及白介素6(IL-6)对角朊细胞与真皮成纤维细胞之间相互作用的影响以及前列腺素E₁(PGE₁)的调节作用.IL-1α主要由角朊细胞产生,而IL-6则主要由成纤维细胞产生.来自成纤维细胞的培养上清液有较强的促进角朊细胞增殖能力.来自角朊细胞的培养上清液也有一定促进成纤维细胞增殖能力.IL-1α和PGE₁可促进成纤维细胞IL-6的产生.抗IL-6抗体可抑制成纤维细胞培养上清液对角朊细胞的促生长活性.以上结果说明IL-1α和IL-6可能是介导表皮-真皮相互作用的重要因子.     

重组人甲状旁腺素(1-34)对角质形成细胞分泌IL-6和IL-8的影响

目的研究人甲状旁腺素(PTH)(1-34)对角质形成细胞分泌IL-6和IL-8的影响,探讨其治疗银屑病的分子机理。方法体外培养人角质形成细胞株(HaCaT),研究中设空白对照组、骨化三醇组、骨化三醇和PTH(1-34)联合组以及1.0×10-7mol/L～1.0×10-10mol/L4个不同剂量的PTH(1-34)给药组。48h后收集上清液,用酶联免疫吸附法(ELISA)检测HaCaT细胞分泌至培养液中IL-6和IL-8的含量。结果在1.0×10-7mol/L～1.010-10mol/L四个梯度浓度PTH(1-34)及联合骨化三醇处理48h后,HaCaT细胞分泌IL-6量和空白对照组、骨化三醇组比较,差异无统计学意义,而IL-8的分泌量明显降低(P<0.05),且1.0×10-7mol/LPTH(1-34)处理组IL-8分泌量明显低于其他各组。结论PTH(1-34)治疗银屑病的作用机制可能部分通过调节IL-8的表达和分泌来实现。     

溶菌酶对体外培养的人成纤维细胞基质金属蛋白酶-1和12及赖氨酸氧化酶基因表达的影响

目的 探讨溶菌酶对体外培养的人成纤维细胞基质金属蛋白酶-1和12及赖氨酸氧化酶基因表达的影响。方法 采用酶消化法进行体外人皮肤成纤维细胞原代培养,然后将不同浓度的溶菌酶（0,1 × 10-8,1 × 10-7 mol/L）加入体外培养的人皮肤成纤维细胞中,待药物作用后,提取总RNA,通过逆转录反应获得cDNA并进行体外扩增,对扩增产物进行琼脂糖凝胶电泳,根据条带的平均光密度A值的比值来判断人成纤维细胞中基质金属蛋白酶（MMP）-1、MMP-12及赖氨酸氧化酶（LOX）mRNA的表达水平。结果 对体外培养的人成纤维细胞进行溶菌酶干预,经β肌动蛋白内参校正后,RT-PCR示对照组、低剂量组、高剂量组MMP-1、MMP-12 mRNA表达水平三组间差异具有统计学意义（F值分别为6.98和4.44,P值均 < 0.05）。SNK-q检验显示,低剂量组与对照组、高剂量组比较,差异均无统计学意义（P > 0.05）,高剂量组与对照组相比,差异有统计学意义（P < 0.05）。LOX mRNA表达水平三组间差异具有统计学意义（F = 5.24,P < 0.05）,SNK-q检验显示,低剂量组、高剂量组与对照组相比,差异具有统计学意义（P < 0.05）,低剂量组与高剂量组相比,差异无统计学意义（P > 0.05）。结论 溶菌酶可以下调MMP-1和MMP-12及上调LOX基因的转录水平。     

TGF-β1对人工皮肤光损伤炎症因子IL-1α、IL-6、TNF-α影响的研究

目的探讨TGF-β1对人工皮肤光损伤中炎症因子IL-1α、IL-6、TNF—α的影响。方法将角质形成细胞和成纤维细胞接种到胶原支架中体外构建人工皮肤,将人工皮肤分为四个实验组,即阴性对照组,紫外线照射,TGF-β1干预组,以及TGF—β1干预后再进行UV照射,采用ELISA法检测各实验组上清液中炎症因子IL-1α、IL-6、TNF—α．的浓度变化。结果体外培养的人工皮肤分为表皮和真皮层,与正常皮肤结构相似。与阴性对照组相比,UV照射组炎症因子IL-1仪、IL-6、TNF—α明显增加,有显著性差异（p〈0．01）;TGF-β1对IL-1α、IL-6、TNF-α的浓度没有明显影响（P〉0．05）;但是TGF—β1干预的uV组跟UV照射组比较则是IL—1α、IL-6、TNF-α浓度增加减少（p〈0．01）。结论TGF-β1对紫外线所致的人工皮肤光损伤的炎症因子IL-1α、IL-6、TNF-α产生有一定抑制作用,即对光损伤模型有保护作用。     

尖锐湿疣患者细胞免疫功能和血清可溶性粘附分子水平研究

为了解尖锐湿疣(CA)患者的细胞免疫功能和血清可溶性粘附分子水平,采用ELISA方法检测了33例CA患者血清中IL-2及其可溶性受体(sIL-2R)、IL-6、可溶性细胞间粘附分子-1(sICAM-1)和血管细胞粘附分子-1(sVCAM-1)水平。结果发现患者血清中IL-2及IL-6水平明显低于正常对照(P<0.01),sIL-2R、sICAM-1、sVCAM-1明显高于对照(P<0.05),提示CA患者存在细胞免疫缺陷和血清高粘附分子水平。     

寻常性银屑病患者血脂、TXB_2、6-酮-PGF_(1α)和血小板聚集功能的检测

银屑病患者伴发心血管疾病者明显增多，为探讨其机理，对１００例作血脂检测，发现ＴＣ、ＴＧ和β ＬＰ均明显增高。３２例作ＴＸＢ２和６ 酮 ＰＧＦ１α检测，前者为正常人的２．５倍，后者为１．８倍。７５例作血小板聚集功能检测，也明显增高。提示高脂血症、ＴＸＢ２和６ 酮 ＰＧＦ１α增高以及血小板聚集功能增高是银屑病伴发心血管疾病的病理基础     

A hypomorphic Egfr allele does not ameliorate the palmoplantar keratoderma caused by SLURP1 deficiency

Mutations in SLURP1, a secreted protein of keratinocytes, cause a palmoplantar keratoderma (PPK) known as mal de Meleda. Slurp1 deficiency in mice faithfully recapitulates the human disease, with increased keratinocyte proliferation and thickening of the epidermis on the volar surface of the paws. There has long been speculation that SLURP1 serves as a ligand for a receptor that regulates keratinocyte growth and differentiation. We were intrigued that mutations leading to increased signalling through the epidermal growth factor receptor (EGFR) cause PPK. Here, we sought to determine whether reducing EGFR signalling would ameliorate the PPK associated with SLURP1 deficiency. To address this issue, we bred Slurp1‐deficient mice that were homozygous for a hypomorphic Egfr allele. The hypomorphic Egfr allele, which leads to reduced EGFR signalling in keratinocytes, did not ameliorate the PPK elicited by SLURP1 deficiency, suggesting that SLURP1 deficiency causes PPK independently (or downstream) from the EGFR pathway.     

Dihydroavenanthramide D inhibits mast cell degranulation and exhibits anti‐inflammatory effects through the activation of neurokinin‐1 receptor

Chronic pruritus is difficult to treat. Current treatment options are frequently ineffective and new therapeutic approaches are urgently needed. Avenanthramides are active substances in oats that exhibit anti‐inflammatory effects. Their potential to interrupt pruritus mechanisms was investigated in this study. It was found that the synthetic analog dihydroavenanthramide D (DHAvD) can interact with the neurokinin‐1 receptor (NK1R) and inhibit mast cell degranulation. DHAvD also affects inflammatory processes and reduces secretion of the cytokine interleukin‐6. Our findings indicate that DHAvD may act as a NK1R inhibitor and could be a promising candidate for topical treatments of chronic pruritus.     

Detection of inflammatory cytokines in psoriatic skin

Recent investigations have revealed the involvement of cytokines in the pathogenesis of psoriasis. This study examined the amount of inflammatory cytokines — interleukin-1 (IL-1), interleukin-6 (IL-6) and granulocyte macrophage colony-stimulating factor (GM-CSF) — released into the supernatants of organ cultures of involved and uninvolved skin from psoriatic patients and normal skin from healthy individuals. Bioassays were employed to detect the activities of IL-1 and IL-6. Enzyme-linked immunosorbent assay (ELISA) methods were used to quantitate immunoreactive IL-1, IL-1, IL-6 and GM-CSF. The activity of IL-1 in uninvolved psoriatic skin was found to be increased relative to that in involved and normal skin, while immunoreactive IL-1 was found only in involved skin. A neutralization experiment showed that bioactive IL-1 was mostly attributable to IL-1. Uninvolved psoriatic skin also secreted higher amounts of both bioactive and immunoreactive IL-6 compared with involved skin. Immunoreactive GM-CSF was detected in uninvolved skin only. These cytokines detected in uninvolved skin may have been released from epidermal or mesenchymal cells, since uninvolved skin contained fewer inflammatory infiltrates. Our results offer additional evidence that increased amounts of inflammatory cytokines in uninvolved skin may provide a preliminary condition and play important roles in the initial events in the evolution of psoriatic lesions.Part of this work was presented in abstract form at the Fifth International Psoriasis Symposium in San Fransisco, 10–14 July 1991     

Polymorphisms in inflammation genes (angiotensinogen, TAP1 and TNF-β) in psoriasis

Abstract This study focused on the association between plaque psoriasis and polymorphisms of several inflammation genes. Included in the study were 142 Caucasian (Czech) patients with plaque psoriasis and 141 healthy subjects. The genotypes of the polymorphisms in angiotensinogen [M235T ATG, A(-6)G ATG], in transporters associated with antigen processing TAP1 (TAP1*0101, TAP*02011 and TAP1*0301) and in lymphotoxin α (TNFβ) (NcoI in intron 1) were detected by polymerase chain reaction-based methods and restriction enzyme analysis. An increase in B1 (less frequent) allele of NcoI TNFβ polymorphism was found in psoriatic patients compared to healthy individuals (odds ratio = 1.6, 95% confidence interval 1.13–2.26, P = 0.006). A positive family history of psoriasis was associated with a higher B1 allele frequency in NcoI TNFβ (P = 0.011). Hardy-Weinberg disequilibrium was found in TAP1 polymorphism A→G at nucleotide 1207 in psoriatic patients. A case-control difference was found in the allelic concurrence of M235T and A(-6)G ATG polymorphisms. The most frequent population genotypes MMGG, MTAG and TTAA were observed in 92% of patients vs 74% of control subjects (odds ratio 0.29, 95% confidence interval 0.14–0.60, P = 0.0003). A positive history of tonsillitis and/or tonsillectomy was associated with a higher T allele frequency of the M235T ATG polymorphism (P = 0.037) as well as with a higher G allele frequency of the A(-6)G ATG polymorphism (P = 0.022). Polymorphisms in proinflammatory angiotensinogen and TNFβ genes were associated with plaque psoriasis, a positive family history of psoriasis and with frequent tonsillitis in childhood. Received: 29 February 2000 / Revised: 14 April 2000 / Accepted: 25 August 2000     

靛玉红对角质形成细胞中促炎细胞因子表达水平的影响

 目的: 探讨靛玉红对人类永生化角质形成细胞株（HaCaT细胞）中促炎细胞因子IL-1β、IL-6、TNF-α表达水平的影响，初步阐明青黛膏治疗银屑病的机制。 方法：以人类永生化角质形成细胞株HaCaT细胞作为研究对象，观察不同浓度靛玉红处理后HaCaT细胞中促炎细胞因子IL-1β、IL-6、TNF-α表达水平的变化，采用ELISA检测各组细胞培养上清液中IL-1β、IL-6、TNF-α的分泌水平，RT-qPCR检测各组细胞中IL-1β、IL-6、TNF-α的mRNA表达水平。 结果：靛玉红处理后，HaCaT细胞中促炎细胞因子IL-1β、IL-6、TNF-α mRNA表达水平及细胞培养上清液中三者分泌水平均明显降低，差异均具有统计学意义(P值<0.05)。随着靛玉红浓度升高，该抑制作用增强，呈一定的浓度依赖性。 结论:青黛膏的主要成分靛玉红可能通过降低HaCaT细胞中IL-1β、IL-6、TNF-α等银屑病相关炎性因子的表达水平从而发挥治疗银屑病的作用。     

Modulation of MHC class II+ Langerhans cell numbers in corticosteroid treated epidermis by GM-CSF in combination with TNF-α

Abstract It is important to understand how dendritic cells (DC) are recruited, maintained and stimulated to migrate from tissues to lymph nodes. This is because DC are potent initiators of primary immune responses and candidates for vaccine development. Identification of factors which could lead to increased numbers of DC in tissues could affect immune responses by modulating their interaction with antigen which penetrates the tissue. To identify cytokines which could increase DC in tissues we tested the ability of GM-CSF, TNF-α and IL-6 to partially prevent steroid depletion of Langerhans cells (LC) from the epidermis. Cytokines diluted in serum-containing medium were compared with cytokines diluted in albumin-containing, serum-free medium in order to determine a minimum combination of cytokines required to increase LC and the effect of serum on the LC-increasing activity of cytokines. In the presence of serum, GM-CSF or TNF-α could increase LC frequency compared to the control; but in the absence of serum neither of these cytokines were effective unless they were combined with each other. In the presence of serum the combination of GM-CSF with TNF-α was ineffective. The data support the hypotheses that GM-CSF and TNF-α are both important in regulating LC numbers in the epidermis in vivo. Serum may modulate how each of these cytokines, separately or in combination, affect LC frequency in the epidermis–GM-CSF and TNF-α separately probably interact with other factors present in serum to increase LC frequency, whereas in combination it is possible that these separate effects are cancelled in the presence of serum. TNF-α and GM-CSF together, in the absence of serum, form one combination of a minimum number of cytokines which can regulate LC frequency in the epidermis; and IL-6 alone, or in combination with GM-CSF, does not increase LC frequency.     

白念珠菌对人血管内皮细胞表面黏附分子表达和细胞因子分泌的影响

目的 观察白念珠菌对人血管内皮细胞表面细胞间黏附分子-1(ICAM-1)、血管细胞黏附分子-1(VCAM-1)表达及分泌白介素6(IL-6)、白介素8(IL-8)水平的影响.方法 分离健康产妇胎盘脐静脉内皮细胞进行原代培养,细胞长至单层时,与白念珠菌共培养不同时间,再用逆转录-聚合酶链反应法分析内皮细胞表面ICAM-1mRNA、VCAM-1mRNA的表达,用ELISA法检测IL-6、IL-8分泌水平.结果 白念珠菌刺激4h后血管内皮细胞表达ICAM-1 mRNA、VCAM-1 mRNA明显增加,至8h达到峰值.而IL-6,IL-8的分泌在4h后也明显增加(P<0.05),24h到达高峰(P<0.01).结论 白念珠菌可诱导人血管内皮细胞ICAM-1 mRNA、VCAM-1 mRNA表达及促进IL-6、IL-8分泌.