特应性皮炎皮损角质形成细胞TLR2和TLR4的表达

目的:探讨Toll样受体(TLR)2和TLR4在特应性皮炎(AD)表达及发病机制中的意义。方法:采用免疫组织化学法检测9例中至重度AD患者急性发作期炎性皮损TLR2和TLR4表达。结果:免疫组化检测显示在正常皮肤基底细胞层角质形成细胞(KC)胞膜散在表达TLR2和TLR4。TLR2和TLR4均在AD急性发作期炎性皮损KC胞膜及胞浆过度表达,以膜表达为主,主要定位于基底细胞层至棘细胞层。结论:AD皮损角质形成细胞TLR2和TLR4过度表达可能与AD皮肤天然免疫炎症反应有关。     

湿疹及皮炎

20050989 特应性皮炎皮损角质形成细胞NF-κB表达及外用他克莫司对其的影响/谢志强(北京大学第一医院皮肤科),刘玲玲,窦侠…//北京大学学报.-2004,36 (5).-487—490 采用免疫组织化学二步法检测9例中至重度AD 患者急性发作期炎性皮损与正常皮肤角质形成细胞(KC)NF-κB的表达方式,以及外用0.1%(质量分数)     

他克莫司对银屑病皮损中蛋白酶活化受体2表达的影响

目的:明确他克莫司对寻常型银屑病皮损中蛋白酶活化受体2(PAR-2)的影响。方法:免疫组化法(SP法)检测30例寻常型银屑病皮损外用0.1%他克莫司软膏治疗前后及25名正常对照PAR-2的表达情况。结果:银屑病患者皮损内角质形成细胞PAR-2的表达明显高于正常对照;经0.1%他克莫司软膏外用治疗2周后表达明显下降。结论:PAR-2在皮损角质形成细胞内过度表达可被他克莫司软膏抑制。     

他克莫司对特应性皮炎皮损中蛋白酶活化受体2表达的影响

目的 观察蛋白酶活化受体2（PAR-2）在特应性皮炎（AD）患者皮肤中的表达以及他克莫司对其表达的影响。方法 比较6例中重度AD患者外用0.1%他克莫司软膏治疗3周前后的临床症状、体征变化,切取患者治疗前后皮损及外观正常的皮肤,6例正常皮肤作对照,免疫组化方法检测皮肤PAR-2的表达变化。结果 PAR-2在皮肤的表皮全层尤其是颗粒层以及毛囊、汗腺、血管内皮细胞、神经纤维样结构均表达。AD患者皮损内角质形成细胞PAR-2染色的平均吸光度为4339.6 ± 115.8,与非皮损部位（4189.0 ± 228.9）和正常对照皮肤（3864.0 ± 237.3）比较,其表达明显升高（t = 2.85,P < 0.05;t = 4.31,P < 0.05）;0.1%他克莫司软膏治疗3周后原皮损处角质形成细胞PAR-2染色的平均吸光度为3942.4 ± 176.6,表达较治疗前明显下降（t = 4.55,P < 0.05）;PAR-2的表达水平与AD患者的瘙痒VAS评分、EASI指数和IGA评分均呈正相关。 结论 PAR-2在AD患者皮损角质形成细胞内的表达增加,表达水平与瘙痒程度和皮损严重程度呈正相关,PAR-2在皮损角质形成细胞内的过度表达可被他克莫司抑制。     

特应性皮炎患者皮损中热休克蛋白60、Toll样受体4和核因子-KB p65的表达

目的:研究热休克蛋白60(HSP60)、Toll样受体4(TLR4)、核因子-kB p65(NF-KB p65)在特应性皮炎(AD)患者皮损中的表达水平及其与疾病发生的关系.方法:采用SP免疫组化法检测17例AD患者急性或亚急性期炎性皮损及12份正常皮肤石蜡标本中HSP60、TLR4、NF-kB p65的表达.结果:HSP60、TLR4、NF-kB p65在重度患者皮损角质形成细胞中均过度表达,与正常对照及中度组比较,差异有统计学意义(P＜0.05);HSP60及NF-kB p65在中度患者组中的表达明显高于正常对照,差异具有统计学意义(P＜0.05);HSP60、TLR4、NF-kB p65在AD皮损中的表达水平与湿疹面积及严重度指数(EASI)评分间存在正相关(P＜0.05).结论:HSP60、TLR4、NF-kB p65在AD患者皮损中存在过度表达.HSP60的表达上调可能与AD发病有关.     

NB-UVB联合他克莫司软膏治疗面部脂溢性皮炎疗效评价

目的:评价NB-UVB联合他克莫司软膏治疗面部脂溢性皮炎的疗效及安全性.方法:140例面部脂溢性皮炎患者随机分为2组,治疗组70例采用NB-UVB照射联合他克莫司软膏外用;对照组70例,外用他克莫司软膏,每日2次,治疗4周后评估疗效及不良反应.结果:治疗组的总有效率为94.29％,明显高于对照组(77.14％),差异有统计学意义(P＜0.05).两组均未见明显不良反应.结论:NB-UVB照射联合他克莫司软膏外用治疗面部脂溢性皮炎安全有效.     

他克莫司软膏治疗儿童特应性皮炎疗效和安全性的meta分析

目的 :系统评价他克莫司软膏治疗儿童特应性皮炎的有效性及安全性。方法 :搜索电子数据库MEDLINE and PubMed、the Cochrane Central Register of Controlled(CENTRAL)、EMbase、中国知网、万方数据库以及维普数据库,检索时间从建库到2016年9月。检索他克莫司软膏治疗儿童特应性皮炎的随机对照试验(RCT)并根据Cochrane系统评价方法评价纳入文献质量,提取相关数据并用Stata/SE11.0软件进行meta分析。结果:最终纳入18个RCT共计2821例中到重度儿童特应性皮炎儿童患者。结果显示:(1)0.03%和0.1%他克莫司软膏可以有效治疗儿童特应性皮炎且无明显不良反应;(2)他克莫司软膏疗效优于氢化可的松软膏,但与甲泼尼龙和氟轻松相比差异无统计学意义;(3)0.1%与0.03%他克莫司软膏疗效及不良反应发生率差异无统计学意义。结论:外用他克莫司软膏可有效治疗儿童特应性皮炎,且具有较低的不良反应发生率。     

长期外用他克莫司软膏对特应性皮炎患者免疫力的影响

目的:探讨特应性皮炎患者长期外用他克莫司软膏治疗后血药浓度的变化以及对外周血淋巴细胞亚群的影响。方法:使用他克莫司软膏(0.1%)治疗12例特应性皮炎患者,疗程12个月,于治疗前和治疗结束后使用流式细胞术检测外周血中淋巴细胞亚群,治疗一周时和治疗结束后使用ELISA检测患者他克莫司血药浓度,比较治疗前后的变化。结果:淋巴细胞亚群的变化在治疗前后比较,差异均无统计学意义(P值均>0.05);治疗1周时,他克莫司血药浓度为(1.73±0.48)ng/mL,治疗12个月后浓度为(1.07±0.42)ng/mL,两者差异具有统计学意义(t=16.85,P<0.05);他克莫司软膏使用总量与Th/Ts比值变化量无相关性(r=-0.40,P>0.05)。结论:特应性皮炎长期外用他克莫司软膏存在低水平的系统吸收,他克莫司低水平系统暴露对特应性皮炎患者的外周血淋巴细胞亚群无影响。     

他克莫司软膏治疗成人特应性皮炎对金黄色葡萄球菌定植的影响

他克莫司（VKS06）是一种大环内酯类免疫抑制剂。大量临床证据显示其外用治疗特应性皮炎（AD）疗效显著，临床应用过程中与药物相关的副作用主要表现为表皮的烧灼感等，然而皮肤感染是局部外用免疫抑制剂的潜在并发症。国内外研究均证明AD患者皮损存在严重的金黄色葡萄球菌（简称金葡菌）的定植，甚至在缺乏临床感染的征相或正常皮肤上亦存在金葡菌的定植。因为他克莫司在体外没有抗菌活性，依局部免疫抑制机制外用VKS06可增加金葡菌的定植，因此我们设计了一个可行性研究以观察外用0．1％他克莫司软膏对金葡菌定植的影响。     

他克莫司软膏治疗成人中、重度特应性皮炎的临床研究

目的:比较0.1%,0.03%他克莫司软膏与赋形剂治疗中、重度特应性皮炎(AD)的疗效和安全性.方法:采用随机、双盲、赋形剂平行对照临床研究方法,入选病例按1:1:1比例分为三组,分别随机接受0.1%,0.03%他克莫司软膏或赋形剂治疗,每天2次外搽患处,共3周.结果:有效率:他克莫司软膏0.1%组和0.03%组分别为88.9%和87.5%,赋形剂组为25%;治愈率:他克莫司软膏0.1%组和0.03%组分别为55.6%和50%,赋形剂组为25%,差异有统计学意义(P＜0.05).结论:他克莫司软膏(0.1%和0.03%)治疗成人中、重度特应性皮炎疗效好,安全和耐受性均良好.     

Expression of 27 KD, 65 KD and 72/73 KD heat shock protein in atopic dermatitis: comparison with those in normal skin and contact dermatitis

The expression of Heat Shock Protein (HPS) 72/73, HSP65 and HSP27 in skin lesions of atopic dermatitis (n = 21) was studied and compared with that in contact dermatitis (n = 18) and normal skin (n = 9). Keratinocytes in the whole epidermis expressed both HSP65 and HSP72/73 with a membranous, cytoplasmic or nuclear/perinuclear staining pattern much more intensely in atopic dermatitis than in contact dermatitis and normal subjects. In approximately half of the subjects with atopic dermatitis, infiltrating cells in the dermis expressed HSP65 and HSP72/73; this was not observed in contact dermatitis. HSP27 was expressed in the upper epidermis with a cytoplasmic or nuclear/perinuclear staining pattern in all groups. HSP27 was not expressed by infiltrating cells. A clinical evaluation of atopic dermatitis showed that more severe types of atopic dermatitis expressed more intense expression of HSP65 and HSP72/73, but not HSP27, in their skin lesions. These findings suggested that HSP65 and HSP72/73 may play roles in the pathogenesis of atopic dermatitis.     

Expression of adhesion molecules in atopic dermatitis is reduced by tacrolimus, but not by hydrocortisone butyrate: a randomized immunohistochemical study

Topical tacrolimus represents an effective and well-tolerated treatment for atopic dermatitis (AD). Its known effects include reduced production of proinflammatory cytokines and reduced chemokine gradient. We performed lesional skin biopsies on adult patients affected by moderate-to-severe AD. Then, patients were randomized to receive local treatment with tacrolimus ointment 0.1% and hydrocortisone butyrate ointment 1%. On the 21st day of treatment, another skin specimen was taken. Nine patients treated with tacrolimus and seven treated with hydrocortisone successfully concluded the trial. By immunohistochemistry (alkaline phosphatase/antialkaline phosphatase method), we demonstrated that endothelial leucocyte adhesion molecule (ELAM)-1, vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 showed different intensities and patterns of expression in untreated AD lesions. Tacrolimus-treated specimens featured a significant reduction of the expression of ELAM-1, VCAM-1 and ICAM-1, while hydrocortisone-treated lesions did not. Inhibition of adhesion molecule expression may represent another selective mechanism of action of topical tacrolimus in AD.     

Wet-wrap treatment using dilutions of tacrolimus ointment and fluticasone propionate cream in human APOC1 (+/+) mice with atopic dermatitis

Background  Wet-wrap treatment (WWT) with diluted topical steroids is widely used in atopic dermatitis (AD). Mice with transgenic overexpression of human apolipoprotein C1 (APOC1) in the liver and the skin are not only characterized by hyperlipidaemia and raised IgE levels, but also by pruritic dermatitis and a disturbed skin barrier function, providing a novel in vivo mouse model for AD.
Objectives  We investigated an adapted WWT method in the AD model in APOC1 mice in order to establish its efficacy.
Methods  The effect of topical 0·1% and 0·03% tacrolimus ointment, tacrolimus base ointment, different dilutions of 0·05% fluticasone propionate (FP) cream and emollient on the development of dermatitis in APOC1 mice was investigated. WWT was performed with 0·03% tacrolimus ointment or 0·017% FP cream.
Results  AD in APOC1 mice responded to topical treatment with tacrolimus or FP. In contrast to tacrolimus treatment, FP treatment was associated with loss of body weight. WWT reinforced several therapeutic aspects, notably improvements in transepidermal water loss and in epidermal thickness. WWT using tacrolimus 0·03% ointment was more effective than WWT using FP 0·017% cream.
Conclusions  AD in APOC1 mice responds to treatment with (diluted) tacrolimus or FP; treatment with FP cream, but not tacrolimus ointment, was associated with weight loss. In this study, the adapted WWT using tacrolimus or FP in mice had a limited improving effect as compared with open application of tacrolimus or FP.     

Epidermal caspase-3 cleavage associated with interferon-gamma-expressing lymphocytes in acute atopic dermatitis lesions

Keratinocyte apoptosis mediated by Fas/Fas ligand molecular interactions and subsequent caspase activation is believed to play an important role in the pathogenesis of atopic dermatitis (AD), in particular for the formation of spongiosis. To estimate epidermal caspase activation in normal and AD skin under in vivo conditions, we analysed caspase-3 cleavage by immunohistology. In normal skin as well as non-lesional AD skin, we detected caspase-3 cleavage in single cells of the basal layer. In contrast, in acute lesional AD skin, we not only obtained evidence for increased expression of cleaved caspase-3 in keratinocytes of the basal layer but also observed caspase-3 cleavage in one or more layers of the spinous cell layer, in particular in spongiotic areas. Short-term topical treatment of the skin lesions with tacrolimus or pimecrolimus abolished the expression of cleaved caspase-3 in the spinous layer. Moreover, epidermal caspase-3 cleavage correlated with the numbers of dermal interferon-gamma (IFN-gamma)-expressing CD4+ and CD8+ lymphocytes in skin lesions of AD patients, supporting the view that IFN-gamma is important for the activation of proapoptotic pathways in keratinocytes. This is also confirmed by the observation of increased Fas expression on keratinocytes in acute AD lesions that was markedly reduced following topical calcineurin inhibitor treatment. These data suggest that caspase-3 cleavage in the spinous layer of the epidermis is a pathologic event contributing to spongiosis formation in AD, whereas cleavage of caspase-3 in basal cells might represent a physiologic mechanism within the process of epidermal renewal.     

An open study of a lotion formulation to improve tolerance of tacrolimus in facial atopic dermatitis

BACKGROUND: We identified 19 patients with facial atopic eczema who failed to respond to tacrolimus (FK506) ointment, although tacrolimus ointment has shown excellent benefit for the treatment of recalcitrant facial erythema in most patients with atopic dermatitis. OBJECTIVES: We attempted to determine the efficacy of an original lotion formulation of tacrolimus for facial atopic dermatitis resistant to tacrolimus ointment. PATIENTS/METHODS: Recalcitrant facial erythema of these 19 patients was treated with an original tacrolimus lotion preparation for 6 months. Patch testing with white petrolatum was performed in both the 19 patients and in 30 other atopic dermatitis patients who had experienced excellent results with tacrolimus ointment. RESULTS: Of the 19 resistant patients, those whose symptoms were greatly or moderately improved by the lotion were 95%, 89% and 89% after 2 weeks, 3 months and 6 months of treatment, respectively. Further, patch testing to petrolatum showed positive reactions in several (six of 19) patients, compared with none of 30 controls with atopic eczema that had responded to topical tacrolimus ointment. CONCLUSIONS: The tacrolimus lotion had a significant effect on the recalcitrant facial erythema in adult patients with atopic dermatitis who were resistant to tacrolimus ointment. We suggest that one reason for the unresponsiveness to tacrolimus ointment may be because of contact sensitivity to white petrolatum.     

Tacrolimus clinical studies for atopic dermatitis and other conditions

The first topical immunomodulator approved for human use, tacrolimus ointment (Protopic, Fujisawa, Healthcare, Inc, Deerfield, IL), has been shown to be effective and safe in the treatment of children (aged 2 years and older) and adults with atopic dermatitis (AD). Clinical trials conducted worldwide have involved 12,000 patients, with safety and efficacy data available for up to 3 years of treatment. In addition to its beneficial effects in the management of AD, topical tacrolimus has also been reported to be of benefit in other immunologically mediated skin diseases including: hand dermatitis, contact dermatitis, eyelid dermatitis, erosive lichen planus, steroid-induced rosacea, pyoderma gangrenosum, and graft-versus-host disease. This article reviews the clinical experience of topical tacrolimus in the treatment of AD and other skin conditions.     

Effects of tacrolimus ointment on facial eruption, itch, and scratching in patients with atopic dermatitis

The action of tacrolimus ointment on pruritus in atopic dermatitis is still unclear. In this open study we investigated both the relationship between the severity of eruptions and the degree of itch and scratching in patients with atopic dermatitis and the effects of topical tacrolimus on these symptoms. Seventy adults with moderate to severe atopic dermatitis with facial eruptions that were recalcitrant to topical steroids applied a 0.1% tacrolimus ointment twice per day after discontinuation of topical steroid. The eruption scores and an assessment of the itch and scratching were recorded for 12 weeks. Oral antihistamine was prescribed at least one month before the study and continued unchanged during the study in each patient. The percentage reduction in the score of itch and scratching after two weeks (n=59) was significantly higher than in the score of eruption. Although there was no significant relationship between the severity of the eruptions and the degree of itch and scratching during steroid application, a relationship became significant after four weeks (n=59) of tacrolimus use by a one-factor ANOVA analysis. This suggests that tacrolimus ointment is effective for the itch and scratching in cases where degrees might be discrepant from the severity of eruptions in patients with recalcitrant facial eruptions of AD.