白细胞介素8在银屑病发病机理中的作用

用酶免疫组化法１３例银屑病皮撷 、９例非皮损皮肤和１０例正常皮肤组织中ＩＬ－８的表达与分布，并研究ＩＬ－８对表皮细胞增殖的影响。结果表明９例皮损组织 表层层显著显示ＩＬ－８阳性反应，而患者非皮损皮肤及正常人皮肤表皮层均呈阴性反应。体外实验证实，ＩＬ－８能明显促进原代培养的人表皮细胞增国殖，并呈一定的剂量依赖性。     

Alterations of glucosylceramide-beta-glucosidase levels in the skin of patients with psoriasis vulgaris

Hydrolysis of glucosylceramides by the enzyme glucosylceramide-beta-glucosidase (GlcCer'ase) results in ceramide, a critical component of the intercellular lamellae that mediates the epidermal permeability barrier. A disturbance of ceramide formation is supposed to influence the transepidermal water loss in common skin diseases like atopic eczema or psoriasis. The aim of this study was to investigate whether GlcCer'ase levels were altered in the skin of subjects with psoriasis vulgaris. Skin punch biopsies were taken from lesional and non-lesional psoriatic skin and GlcCer'ase was evaluated both at the RNA and at the protein level. Normal skin from surgical patients provided the baseline GlcCer'ase expression in healthy subjects. Our results show that GlcCer'ase mRNA expression was decreased in psoriatic non-lesional skin compared to normal controls in all cases. Interestingly, in lesional psoriatic skin the level of GlcCer'ase was increased compared to non-lesional skin in all cases. For the immunohistochemical analysis, we used a newly synthesized monoclonal antibody anti-human GBC (GlcCer'ase-GST fusion protein). The results confirmed that GlcCer'ase, mainly present in the upper epidermis, was decreased in psoriatic skin compared to normal control and was increased in lesional compared to non-lesional psoriatic skin. Our findings support the concept that alteration in water permeability barrier in lesional psoriatic skin can serve as a trigger for the upregulation of the expression of enzymes like GlcCer'ase with consequent stimulation of ceramide generation.     

寻常性银屑病患者表皮中DNA总体甲基化水平检测

目的通过检测寻常性银屑病患者表皮DNA总体甲基化水平,探讨DNA甲基化在银屑病发生发展过程中的作用。方法采用荧光比色法检测寻常性银屑病患者表皮DNA总体甲基化水平,与同一患者非皮损部位及健康对照者表皮进行对照研究,并分析其与病情严重程度的相关性。以PASI评分评估银屑病病情严重程度。结果在30例银屑病患者皮损、非皮损部位表皮及28名健康对照者表皮中检测到不同程度的DNA甲基化,分别为(4.92±2.12)%、(2.85±1.35)%和(2.32±0.99)%。银屑病患者皮损部位表皮DNA总体甲基化水平显著高于同一患者的非皮损部位(P<0.0001)及健康对照者(P<0.0001),而非皮损部位DNA总体甲基化水平与健康对照者之间差异无统计学意义(P>0.05)。银屑病患者皮损部位表皮中DNA总体甲基化水平与PASI评分之间存在显著相关性(P<0.0001)。结论寻常性银屑病患者表皮中DNA总体甲基化水平明显升高,且与疾病严重程度呈正相关。     

Interleukin-6 in the epidermis of patients with psoriasis before and during PUVA treatment

Biopsies from lesional and unaffected skin of 6 patients with psoriasis, taken before and during treatment with psoralen plus UVA (PUVA) were examined immunohistologically, using partially purified polyclonal antibodies to crude supernatants of activated human blood monocytes. By absorption with recombinant derived human monokines, we were able to demonstrate that interleukin-6 (IL-6) (but not IL-1 alpha or IL-1 beta) was located in a laminar and granular pattern in stratum corneum, and on epidermal cell membranes in the viable cellular epidermis. Before PUVA treatment, the intensity and the extension of staining for IL-6 were both markedly increased in lesional skin compared with uninvolved skin. A weaker staining for IL-6 was observed in lesional skin, simultaneous with the clinical improvement of psoriasis. The staining patterns for IL-6 in biopsies from cleared lesional skin and uninvolved psoriatic skin were identical at the conclusion of therapy.     

银屑病表皮热休克蛋白27、70、60的表达

目的：探讨热休克蛋白（HSP）在银屑病发病中的作用。方法：通过免疫组化和图像分析检测了25例银屑病患者治疗前后皮损、非皮损区表皮组织中HSP27、HSP70和HSP60的表达，并与6例正常人作对照。结果：HSP27、HSP70在非皮损、正常人表皮中呈基础表达，在银屑病患者皮损中的表达很弱，治愈后表皮后HSP27、HSP70的表达又恢复；HSP60的表达在银屑病皮损组表皮中均为阳性，而银屑病非皮损组、治愈后组表皮中及正常人对照表皮组织中HSP60的表达均阴性。结论：热休克蛋白在银屑病应激保护机制中可能发挥一定的作用。     

Lichen planus remission is associated with a decrease of human herpes virus type 7 protein expression in plasmacytoid dendritic cells

The cause of lichen planus is still unknown. Previously we showed human herpes virus 7 (HHV-7) DNA and proteins in lesional lichen planus skin, and significantly less in non-lesional lichen planus, psoriasis or healthy skin. Remarkably, lesional lichen planus skin was infiltrated with plasmacytoid dendritic cells. If HHV-7 is associated with lichen planus, then HHV-7 replication would reduce upon lichen planus remission. HHV-7 DNA detection was performed by nested PCR and HHV-7 protein by immunohistochemistry on lesional skin biopsies from lichen planus patients before treatment and after remission. Biopsies were obtained from lichen planus lesions before treatment (n = 18 patients) and after remission (n = 13). Before treatment 61% biopsies contained HHV-7 DNA versus 8% after remission (P = 0.01). HHV-7-protein positive cell numbers diminished significantly after remission in both dermis and epidermis. Expression of HHV-7 was mainly detected in BDCA-2 positive plasmacytoid dendritic cells rather than CD-3 positive lymphocytes. HHV-7 replicates in plasmacytoid dendritic cells in lesional lichen planus skin and diminishes after remission. This study further supports our hypothesis that HHV-7 is associated with lichen planus pathogenesis.     

银屑病患者皮损中转化生长因子-β1及受体基因表达的研究

目的 研究银屑病患者皮损及非皮损处转化生长因子-β1(TGF-β1)、转化生长因子β受体(TGF-βRⅡ)及CD105的基因(mRNA)表达,并初步探讨其临床意义。方法 采用异硫氰酸胍法抽提皮肤组织中的RNA;采用逆转录聚合酶链反应(RT-PCR)检测皮肤组织中mRNA的表达。结果 银屑病患者皮损组织TGF-β1及TGF-βRⅡmRNA的表达均低于非皮损组织及正常人(P<0.05);非皮损组织及正常人皮肤组织中mRNA的表达量差异无显著性(P>0.05)。而银屑病皮损组织CD105的mRNA表达量高于非皮损组及正常人(P<0.05);非皮损组织和正常人皮肤组织比较P>0.05。结论 银屑病皮损中TGF-β1及TGF-βRⅡ表达降低和CD105的表达上调可能与银屑病的表皮过度增殖及真皮炎症细胞浸润有关。     

Correlation between macroscopic fluorescence and protoporphyrin IX content in psoriasis and actinic keratosis following application of aminolevulinic acid

In fluorescence diagnosis with 5-aminolevulinic acid (ALA)-induced porphyrins (FDAP), protoporphyrin IX (PpIX) accumulation can be macroscopically visualized. Interpretation of these data is still problematic because of the low reproducibility of the procedure and poor understanding of the mechanisms involved in PpIX tumor selectivity. In this study, PpIX accumulation is investigated in patients with psoriasis and actinic keratosis (AK) following FDAP. For this purpose, desquamated lesional and non-lesional skin were incubated with 20% ALA ointment for 3 h, FDAP was performed, and highly fluorescing lesional skin and non-lesional skin were biopsied. In extracts from these biopsies, PpIX, protein, and dsDNA were quantified by spectrofluorometry. Digital images acquired with FDAP were analyzed using image analysis software. PpIX per biopsy in lesional skin in both psoriasis and AK was significantly higher than in non-lesional skin (p < 0.05). When corrected for epidermal involvement, only lesional psoriatic skin showed significantly higher PpIX levels than non-lesional skin. The PpIX-ratio lesional:non-lesional skin (mean(pmol per mL)+/-SEM) was 4.12+/-0.91 in psoriasis and 1.96+/-0.24 in AK. In FDAP, the ratio of lesional:non-lesional skin was 1.77+/-0.06 in psoriasis and 1.37+/-0.07 in AK. Macroscopic fluorescence and PpIX content appeared to be well correlated (r = 0.73), thus making FDAP a good predictor of PpIX content.     

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.     

肝素结合表皮生长因子样生长因子在银屑病早期皮损中的意义

目的：探讨肝素结合表皮生长因子样生长因子（HB－EGF）在银屑病发上期中的作用机制。方法：用原位杂交和免疫组化的方法检测正常组织、进行期银屑病皮损及其未受累皮肤中HB－EGF mRNA的蛋白表达。结果：在正常人皮肤组织中，HB－EGF mRNA和蛋白位于基底层（100％），基底上层仅有少量表达（16．67％）；在未受累表皮和银屑病皮损的周围部分中，HB－EGF的表达不仅位于基底层，且以灶状高表达于基底上层（分别为88．00％、80．00％）；在银屑病皮损的中央部分，基底上层无HB－EGF表达，基底层几乎不表达（4．00％）。结论：HB－EGF可能在银屑病发病的早期阶段起重要作用。     

银屑病皮损中IL—1受体拮抗物的蛋白和mRNA表达

采用抗人ＩＬ－１ｒａ单抗和地高辛标记的ＲＮＡ探针,在银屑病破损和正常人皮肤切片上进行ＡＰＡＡＰ染色和原位杂交,检测并比较ＩＬ－Ｉｒａ的蛋白表达和ｍＲＮＡ转录水平。结果发现,正常人皮肤中ＩＬ－１ｒａ蛋白染色阴性,ＩＬ－１ｒａｍＲＮＡｗ信号微弱;相比之下,银屑病皮损中ＩＬ－１ｒａ蛋白和ｍＲＮＡ的表达均异常增高,统计学比较相差显著。上述结果显示：ＩＬ－１ｒａ在银屑病皮损中增强表达,提示银屑病病理过程中Ｉ     

寻常性银屑病患者表皮中DNA甲基化转移酶2和3a mRNA的表达

目的 检测寻常性银屑病患者表皮中DNA甲基化转移酶（DNMT2和DNMT3a）mRNA表达。方法 利用实时定量PCR技术检测2009年3月至2010年12月来自中国医学科学院皮肤病医院皮肤科及宜兴市人民医院皮肤科门诊的46例寻常性银屑病患者皮损和非皮损表皮以及来自中国医学科学院皮肤病医院皮肤外科的28例健康对照表皮中DNMT2和DNMT3a mRNA的表达。结果 在银屑病患者皮损、非皮损和对照组表皮DNMT2 mRNA表达水平（2-ΔΔCt值）分别是0.62 ± 0.02、0.36 ± 0.05和0.15 ± 0.11,皮损组明显高于非皮损组（t = 6.23,P < 0.01）,非皮损组明显高于对照组（t = 7.33,P < 0.01）;DNMT3a mRNA表达水平（2-ΔΔCt值）分别是0.85 ± 0.03、0.43 ± 0.04和0.18 ± 0.09,皮损组明显高于非皮损组（t = 5.66,P < 0.01）,非皮损组明显高于对照组（t = 8.62,P < 0.01）。结论 寻常性银屑病患者表皮DNMT2和DNMT3a mRNA均异常高表达。     

Immunofluorescence localization of nuclear retinoid receptors in psoriasis versus normal human skin

Psoriasis responds favourably to treatment with retinoids but the cellular pathways mediating these effects are poorly understood. Retinoids regulate keratinocyte proliferation and maturation via binding to nuclear retinoic acid receptors (mainly RARalpha and RARgamma) which form heterodimers with the 9-cis-RA receptor, RXRalpha. We have previously shown that mRNA expression of RARalpha and RXRalpha is down-regulated in psoriatic lesions as compared with non-lesional human skin. In the present study, we investigated the protein expression of RARalpha, RARgamma and RXRalpha in normal and psoriatic skin using indirect immunofluorescence analysis. Epidermal keratinocytes of normal and non-lesional psoriatic skin displayed similar nuclear localization of all three receptors; RARalpha was detected with decreasing intensity from basal to suprabasal layers, RARgamma showed the opposite trend, whereas RXRalpha was evenly expressed throughout the epidermis. In lesional psoriatic skin, however, all three receptor proteins showed a much higher staining intensity in the lower half of the epidermis; in particular, RARalpha immunoreactivity was low or even absent in the upper layers of epidermis. The results support the idea that psoriasis is associated with abnormal retinoid signalling in lesional epidermis.     

Enhanced expression of the high-affinity receptor for IgE (Fc(epsilon)RI) associated with decreased numbers of Langerhans cells in the lesional epidermis of atopic dermatitis

Epidermal Langerhans cells (LCs) and the high-affinity receptor for IgE (Fc(epsilon)RI) on their surface are considered important in the pathogenesis of atopic dermatitis (AD). We investigated the numbers of epidermal LCs and their Fc(epsilon)RI expression in patients with AD and healthy controls. Biopsy specimens taken from lesional skin from 17 patients with AD, non-lesional skin from five patients with AD and normal skin from five healthy individuals were immunohistochemically stained with a monoclonal antibody against CD1a or with either of two monoclonal antibodies against two different epitopes of Fc(epsilon)RI alpha chain. Many dendritic cells were positively stained with anti-CD1a antibody in the epidermis of each skin sample, and fewer cells were stained with anti-Fc(epsilon)RI antibodies. The numbers of epidermal LCs positive for Fc(epsilon)RI were significantly increased in both lesional and non-lesional skin from AD patients compared with those in normal skin, suggesting important roles of Fc(epsilon)RI+LCs in the pathogenesis of the disease. In contrast, the numbers of total epidermal LCs (CD1a-positive) were decreased in AD lesional skin compared with those in non-lesional skin from AD patients and in normal skin from healthy subjects. Together with our finding that the numbers of epidermal LCs were negatively correlated with the clinical severity of the AD lesions, we concluded that epidermal LCs may decrease in some conditions of AD, probably in lesions with severe inflammation.