应用RT—PCR技术检测银屑病皮损中IGF—1受体mRNA

为了了解银屑病皮损中ＩＧＦ－１受体ｍＲＮＡ的表达,采用ＲＴ－ＰＣＲ对２０例银屑病和正常人皮肤组织块进行检测,结果证实银屑病患者表皮中ＩＧＦ－１受体ｍＲＮＡ表达显著高于对照（Ｐ〈０．０１）,真皮中未有ＰＧＦ－１受体与银屑病角度形成细胞在转录水平与增殖,分化有关,ＩＧＦ－１通过旁分泌机制作用于角质形成细胞上受体。     

银屑病患者皮损周围正常皮肤中白介素1系统mRNA原位杂交检测

近来文献报道,表皮白介素 1（ IL－ 1）系统的调节紊乱与银屑病皮损的形成有密切关系 [1]。我们采用原位杂交技术和针对 IL－ 1全部家族成员的 RNA探针,在银屑病患者的非皮损皮肤和正常人皮肤中对 IL－ 1α、 IL－ 1β、 IL－ 1R1（Ⅰ 型受体）、 IL－ 1R2（Ⅱ 型受体）、 IL－ 1ra（受体拮抗剂）的 -----------mRNA转录水平进行了比较,报道如下。 一、材料和方法 皮肤标本来源： 12例银屑病非皮损皮肤均来自寻常型静止期银屑病患者;女 6例,男 6例,年龄 26～ 60岁,在其躯干部皮损 5 cm以外的正常部位取材。 12例正常人皮…     

进行期银屑病患者皮损中胰岛素样生长因子Ⅰ受体蛋白与mRNA的表达

目的 探讨胰岛素样生长因子Ⅰ受体（ＩＧＦ－ＩＲ）和进行期银屑病的关系。方法 应用免疫组织化学逆 酶聚合酶链反应和原位杂交分别检测１５例银屑病患者和１０例正常人皮肤组织块中ＩＧＦＩＲ蛋白与ｍＲＮＡ的表达。结果 ＩＧＦＩＲ蛋白与ｍＲＮＡ表达在银屑病患者基底层和棘细胞层，ＩＧＦＩＲ蛋白与ｍＲＮＡ相对积分密度值在正常对照组与银屑病组间存在极显著性差异。结论进行期银屑病患者皮损中表达增加的ＩＧＦＩＲ蛋白和ｍ     

银屑病皮损VDR表达水平的研究

为了解维生素D受体（VDR）在银屑病皮损中的表达情况,采用RT－PCR方法检测了10例寻求型银屑病患者皮损VDR mRNA表达水平,并与正常人皮肤做对照。结果显示：寻常型银屑病患者皮损VDR mRNA水平明显低于 正常人皮肤（P＜0．05）,寻常型银屑病皮损VDR mRNA水平下降,可能与银屑病的发病有一定的关系。     

IL－15 mRNA在寻常型银屑病皮损中的表达

@@@@目的：检测IL－15 mRNA在寻常型银屑病（PV）皮损中的表达。方法：采用原位杂交方法检测35例寻常型银屑病患者皮损及20例正常人表皮IL－15 mRNA的表达。结果：在正常皮肤组织中IL－15 mRNA表皮基底层阳性着色3例，阳性率15．0％，阴性着色17例；在PV皮损组织中，IL－15 mRNA主要在表皮基底层形成细胞胞浆阳性着色，阳性率100％，强阳性为57．1％（20？35），中强阳性为31．4％（4？35），阳性为11．4％（4？35），两组间有显著性差异。在PV皮损组织中IL－15 mRNA表达线性密度为9．512±6．503，高于在正常皮肤组织中的表达（1．339±1．011），差异有统计学意义。结论：IL－15 mRNA在PV的发病过程中有显著的作用，为阐明PV的发病机制及病变的演变过程提出了新的理论依据。     

白介素1受体拮抗剂基因内含子2多态性与银屑病的关系

为了探讨ＩＬ－１ｒａ内含子２－多态性与银屑病的关系，应用ＰＣＲ确定了１组３０例银屑病嘲笑 １组对照的ＩＬ－１ｒａ内含子２的等位基因频率，发现银屑病组Ａ１、Ａ２和Ａ３的等位基因频率与对照组大致相同，这与国外的结果不同，提示ＩＬ－１ｒａ内含子２可能不是一个与我国银屑发病有关的遗传因素。     

ICAM—1，IGF—1在银屑病皮损中的表达

目的了解银屑病患者皮损中ICAM-1、IGF-1表达水平及分布,并讨论其临床意义和相互关系.方法用免疫组化的方法对16例实验组标本和16例对照组标本进行检则.结果实验组ICAM-1为72.9375±21.1408,对照组为51.3750±17.0876,两组间存在显著性差异(P＜0.01);IGF-1实验组为50.1875±13.5288,对照组为33.3125±17.8281,两组间存在显著性差异(P＜0.02);患者组ICAM-1、IGF-1增高呈明显的正相关(P＜0.05).结论银屑病皮损中ICAM-1、IGF-1表达增高且密切相关,在银屑病病程中起着重要作用.     

银屑病患者皮损组织中角质形成细胞生长因子mRNA的过表达

角质形成细胞生长因子(keratinocyte growth factor,KGF)由Rubin等^[1]在1989年分离鉴定,它特异地作用于上皮细胞,促进其生长增殖和分化,而银屑病的主要特征就是角质形成细胞生长增殖速度过快,因而探讨KGF与银屑病发病的关系,就显得很有意义.我们抽提了12例银屑病皮损的总RNA,进行RNA印迹(Northern blot),结果如下.     

银屑病皮损区IL—8,HLA—DR,PLA2,ANAE,EGFR mRNA,c—fos mRNA的 …

目的 探讨银屑病患者白介素８（ＩＬ－８）,人类白细胞抗原ＤＲ（ＨＬＡ－ＤＲ）,磷脂酶Ａ２（ＰＬＡ２）,酸性α－乙酸萘酯酶（ＡＮＡＥ）,表皮生长因子受体基因（ＥＧＦＲｍＲＮＡ）,ｃ－ｆｏｓ基因（ｃ－ｆｏｓｍＲＮＡ）在皮损的表达定位。方法 对２０例银屑病患者皮损和１０例正常人的皮肤标本进行了免疫组化,组化和原位要交技术检测,结果 ＩＬ－８免疫反应于主要定位于表皮的基层,棘细胞层和颗粒层,血管内皮细胞及     

银屑病患者皮损中IL－2R表达的观察

应用Ｔａｃ单克隆抗体、ＡＢＣ免疫组化技术测定了１８例寻常型患者皮损中的ＩＬ－２Ｒ水平。结果发现：银屑病患者皮损中ＩＬ－２Ｒ水平明显高于正常人；进行期高于静止期。作者认为ＩＬ－２Ｒ是银屑病发病机理中的重要因素之一。     

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.     

Increased expression of adhesion receptors in both lesional and non-lesional psoriatic skin

Adhesion receptors and their ligands play a vital role in the immune system. We studied the expression of different adhesion receptors, using single- and double-staining immunohistochemical techniques, in both lesional and non-lesional skin specimens from seven psoriasis patients and in skin biopsy specimens from eight normal healthy controls. Our results showed an overall increased expression of several adhesion receptors in both lesional and non-lesional psoriatic skin. We consistently found an increased expression in particular of ICAM-1 and E-selectin on endothelial cells, and ICAM-1 on T cells and Langerhans cells. In contrast, a weak expression of VCAM-1 was found on endothelial cells and mononuclear cells in lesional psoriatic skin specimens alone. Interestingly, LFA-1 was also expressed on Langerhans cells, with a greater frequency in skin from lesional than from non-lesional sites, but was never expressed in skin from normal healthy individuals. Furthermore, significantly increased numbers of Langerhans cells and T cells with a positive reactivity for MAb HECA-452 were found in both lesional and non-lesional psoriatic skin. We hypothesize that the enhanced expression of adhesion receptors on migrating immunocompetent cells and endothelial cells of psoriatic skin in general facilitates the increased influx of activated T lymphocytes and other immunocomponent cells into the skin, and thus underscores the generalized character of the disease.     

WAF1／CIP1mRNA在银屑病表皮中的表达

采用原位杂交法研究了ＷＡＦ１／ＧＩＰ１ｍＲＮＡ在正常表皮及银屑病患者表皮中的表达。发现ＷＡＦ１／ＣＩＰ１ｍＲＮＡ在４便正常对照皮肤中不表达，在２８例银屑病患者表皮中呈高表达，ＷＡＡＦ１／ＣＩＰ１ｍＲＮＡ主要在银屑病患者表皮棘细胞层上部和颗粒层细胞中表达，在基底细胞层则基本表达。提示ＷＡＦ１／ＣＩＰ１ｍＲＮＡ在银屑病患者表皮中的高表达可能与银屑病表皮的细胞分化有关。     

IL-1 and IL-1 receptor antagonist regulation during keratinocyte cell cycle and differentiation in normal and psoriatic epidermis

Abstract Changes in the levels of IL-1 (IL-1α, IL-1β, and its receptor antagonist, IL-1RA) occur upon keratinocyte differentiation in vitro and are associated in vivo with abnormal differentiated and hyperproliferative states of psoriatic keratinocytes. A flow cytometric procedure, capable of detecting changes in the intracellular levels of IL-1, was used to determine whether intracellular IL-1/IL-1RA levels in psoriatic and normal keratinocytes alter during in vivo differentiation and the cell cycle. Increases in the IL-1RA levels and IL-1α levels were observed as both normal and psoriatic keratinocytes differentiated from basal stem cells (β₁ integrin⁺, small size) into transient amplifying cells (TAC; β₁ integrin⁺, large size). Upon further differentiation (β₁ integrin^–, large size) both IL-1RA and IL-1α levels dropped. However, while psoriatic IL-1β levels increased as cells differentiated into TACs, little change occurred in the IL-1β levels of normal keratinocytes during differentiation. Changes in IL-1/IL-1RA levels were also detected as keratinocytes progressed through the cell cycle. Within the basal stem cell population of both normal and psoriatic keratinocytes, the IL-1α and IL-1RA levels increased between G0/G1 and S but not between S and G2/M. However, psoriatic basal keratinocyte IL-1β levels differed from those of normal keratinocytes by showing no increase between S and G2/M. The IL-1/IL-1RA levels of normal TAC increased throughout the cell cycle. However, in psoriatic TAC, a slight decrease in IL-1α and IL-1RA levels was observed between G0/G1 and S followed by a delayed increase between S and G2/M. IL-1β levels in psoriatic TAC varied little throughout the cell cycle. Thus, we were able to detect precisely the regulation of IL-1/IL-1RA intracellular levels during the keratinocyte cell cycle and differentiation, showing notably decreased IL-1β upregulation in psoriatic keratinocytes progressing through the cell cycle. Received: 15 July 1996     

寻常性银屑病皮损表皮Dsg1 mRNA的表达

银屑病特征性的病理改变不仅与角质形成细胞(KC)异常增殖有关,也与KC凋亡异常有关[1-2].凋亡是由天冬氨酸特异性半胱氨酸蛋白酶家族(caspase)成员介导的蛋白酶级联反应过程,其中caspase-3是主要的凋亡效应分子.桥粒芯糖蛋白1(Dsg1)是caspase底物蛋白.我们采用RT-PCR方法从mRNA水平检测Dsg1与caspase-3在银屑病皮损中的表达,以探讨Dsg1在银屑病皮损表皮KC凋亡中的作用.     

血管活性肠肽受体1基因在银屑病表皮早期皮损中的表达

目的研究血管活性肠肽受体1(vasoactive intestinal peptide receptor 1 VIPR1)在寻常型进行期银屑病皮损表皮中的表达情况,探讨其在银屑病发病机制中的作用。方法应用组织切片原位杂交法检测VIPR1 mRNA在正常表皮、进行期银屑病皮损表皮和未受累皮肤表皮中的表达和分布。结果在寻常型进行期银屑病皮损表皮的全层、表皮中下层可见VIPR1 mRNA表达(63.6%);在寻常型进行期银屑病非皮损表皮的中下层或个别细胞内可见VIPR1 mRNA的表达(54.5%);在正常皮肤表皮的基底层或少量细胞内可见VIPR1 mRNA的表达(18.2%)。寻常型进行期银屑病皮损表皮中VIPR1 mRNA的表达明显高于正常对照组(P<0.05)。结论VIPR1 mRNA在银屑病发病早期阶段的表达增高可能与银屑病的发病有关。     

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     

银屑病表皮角质形成细胞中白细胞介素8的原位表达

目的 分析ＩＬ－８ｍＲＮＡ在正常健康人表皮及银屑病患者皮损中的表达。方法 用原位杂交（ＩＳＨ）的方法对ＩＬ－８ｍＲＮＡ在正常人表皮及寻常性银屑病不同时期,不同部位表皮的表达进行对照研究。结果 ＩＬ－８ｍＲＮＡ在大多数银屑病皮损标本的表皮层内有特异性杂交信号,而在非皮损区,正常皮肤及临床治愈的银屑病皮肤表皮层无特异性杂交信号。结论 纠正ＩＬ－８在银屑病表皮角质形成细胞中的表达紊乱,可能为该病的防治提