苯并芘与肽聚糖体外协同对人皮脂腺细胞炎症因子表达的影响

目的：检测苯并芘（Bap）和革兰氏阳性菌细菌壁肽聚糖（PGN)体外对SZ95人皮脂腺细胞炎症因子表达的影响。方法：体外培养SZ95永生化人皮脂腺细胞,分为空白对照组、Bap组、PGN组和（Bap+PGN）组, PGN浓度为20μg/mL,Bap浓度 为10-5mol/L。 RT-PCR和ELISA检测SZ95永生化人皮脂腺细胞分泌白介素(IL-1α、IL-1β）、TNF-α mRNA和蛋白表达水平。结果：（Bap+PGN）组IL-1α,IL-1β和TNF-αmRNA及蛋白表达水平高于空白对照组、Bap组和PGN组,差异具有统计学意义（P<0.01）。结论：苯并芘协同增强PGN诱导的皮脂腺细胞炎症因子IL-1α,IL-1β,TNF-α的表达,这可解释为什么环境污染可加重或诱导痤疮 的发生。     

Tumor necrosis factor-α-activated human adipose tissue-derived mesenchymal stem cells accelerate cutaneous wound healing through paracrine mechanisms

Human adipose tissue-derived mesenchymal stem cells (ASCs) stimulate regeneration of injured tissues by secretion of various cytokines and chemokines. Wound healing is mediated by multiple steps including inflammation, epithelialization, neoangiogenesis, and proliferation. To explore the paracrine functions of ASCs on regeneration of injured tissues, cells were treated with tumor necrosis factor-α (TNF-α), a key inflammatory cytokine, and the effects of TNF-α-conditioned medium (CM) on tissue regeneration were determined using a rat excisional wound model. We demonstrated that TNF-α CM accelerated wound closure, angiogenesis, proliferation, and infiltration of immune cells into the cutaneous wound in vivo. To assess the role of proinflammatory cytokines IL-6 and IL-8, which are included in TNF-α CM, IL-6 and IL-8 were depleted from TNF-α CM using immunoprecipitation. Depletion of IL-6 or IL-8 largely attenuated TNF-α CM-stimulated wound closure, angiogenesis, proliferation, and infiltration of immune cells. These results suggest that TNF-α-activated ASCs accelerate cutaneous wound healing through paracrine mechanisms involving IL-6 and IL-8.     

Effects of low concentrations of cis- and trans-urocanic acid on cytokine elaboration by keratinocytes

The urocanic acid cis isomer (cis-UCA) is a possible cutaneous photoreceptor for the immunomodulatory phenomena that follow ultraviolet B irradiation. Several experiments in animals show an inhibitory action of cis-UCA on cellular immunity. However, the action of cis-UCA on the synthesis of cytokines in keratinocytes remains unknown. Long-term cultures of normal human keratinocytes were prepared in a serum-free medium, and stimulated with 1 μg/ml of phorbol 12-myristate 13-acetate (TPA) and UCA or UVB-UCA (10–100 μg/ml). Synthesis of the following cytokines was measured using ELISA and Northern blot techniques: TNF-α, IL-1α, IL-1β, IL-6, IL-8 and TGF-β1. TPA increased TNF-α protein levels in culture supernatants. No changes in IL-1α and IL-1β protein levels were detected in basal culture supernatant after TPA stimulus. TPA augmented RNA expression for TNF-α, IL-1α, IL-1β and TGF-β1. UCA isomers did not induce cytokine changes in protein synthesis. Expression of IL-1α and IL-1β genes was increased after exposure to 100 μg/ml UVB-UCA (70 μg/ml cis-UCA). A slight increase in TNF-α RNA expression was detected when the dose of UVB-UCA reached 100 μg/ml. No effects on cytokine synthesis were found after UCA stimulus. These results suggest that low doses of cis-UCA do not effect cytokine synthesis by keratinocytes.     

Does the presence of bacterial urinary infection contribute to the development of incontinence-associated dermatitis? A scoping review

ObjectiveIncontinence-associated dermatitis (IAD) is an inflammatory skin condition caused by the repeated exposure to urine and faeces. It is not common for urinary incontinence only to cause IAD, however patients with urinary tract infections (UTIs) are also at increased risk for IAD. This scoping review aimed to provide a summary of the relationship between bacterial urinary infections and IAD.MethodsWe conducted a scoping review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews. PubMed, CINAHL, Medline, and Web of Science were searched for relevant articles from January 2007 through February 2020.ResultsBased on eligibility criteria, 13 research studies and review articles were included. Despite the acknowledged role of bacterial infections can play in IAD and the importance of eradicating infections for the prevention of skin breakdown, there have been limited studies that have investigated how uropathogenic bacteria, in combination with urine, lead to skin damage and IAD. The use of urinary catheters also predisposes to UTIs; however, prevalence/incidence rates of IAD in these patients are not clear, as they were considered as continent of urine in the included studies.ConclusionFurther research is needed to elucidate the mechanisms of how bacteria, in combination with urine, lead to IAD.     

TNF-α和IL-1β对HaCaT细胞诱生型一氧化氮合酶表达的影响

目的　探讨炎症性细胞因子肿瘤坏死因子 α(tumornecrosisfactor α ,TNF α)协同白细胞介素 1β(inter leukin 1β ,IL 1β)对体外培养角质形成细胞 (keratinocye ,KC)株HaCaT细胞诱生型一氧化氮合酶 (induciblenitricoxidesyn thase ,iNOS)mRNA和蛋白表达的调节作用 ,以及地塞米松对TNF α和IL 1β作用的影响。 方法　用RT PCR、Westernblotting和免疫组织化学 (SP)方法检测HaCaT细胞iNOSmRNA和蛋白表达情况。结果　正常培养HaCaT细胞iNOS微弱表达或不表达 ,TNF α协同IL 1β显著上调HaCaT细胞iNOSmRNA和蛋白表达 ,地塞米松可显著抑制TNF α和IL 1β的作用。结论　推测TNF α和IL 1β可能通过上调角质形成细胞表达iNOS合成释放的一氧化氮 (niricoxide ,NO)参与皮肤免疫和炎症反应 ;地塞米松的治疗效应可能部分与其能抑制iNOS表达有关。     

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。     

Inter-regulation of Th17 cytokines and the IL-36 cytokines in vitro and in vivo: implications in psoriasis pathogenesis

Accumulating evidence indicates that IL-1 family members and Th17 cytokines have a pathogenic role in psoriasis. We investigated the regulatory interactions of the IL-1-like IL-36 cytokine family and the Th17 cytokines in the context of skin inflammation. We observed increased gene expression of all three IL-36 cytokines in a Th17-dominant psoriasis-like animal model. The induction was downregulated by neutralizing IL-22. Expression of the IL-36s was also induced in cultured primary human keratinocytes (KC) by IL-17A and tumor necrosis factor (TNF)-α, and IL-22 synergized with IL-17A and TNF-α. Furthermore, the IL-36s directly induced their own expression and the production of proinflammatory mediators (TNF-α, IL-6, IL-8) in KC. These functions were markedly enhanced with the addition of IL-17A or TNF-α to the cultures. Similarly, IL-36α and IL-36β augmented IL-17A-mediated induction of antibacterial peptides. Finally, we show that the increased gene expression of IL-36 correlated with Th17 cytokines in the lesions of psoriatic patients. Our results indicate that the IL-36 cytokines are not only regulated by Th17 cytokines, but that they themselves can regulate the expression and enhance the function of Th17 cytokines. We propose that a feedback loop between the IL-36 and Th17 cytokines is involved in driving cytokine expression in psoriatic tissues.     

Effects of interleukins, colony-stimulating factor and tumour necrosis factor on human hair follicle growth in vitro: a possible role for interleukin-1 and tumour necrosis factor-α in alopecia areata

The immune system may be involved in the regulation of normal hair follicle growth as well as in the pathogenesis of some hair diseases. Immunomodulatory cytokines not only act as mediators of immunity and inflammation but also regulate cell proliferation and differentiation and. as such, may play an important part in regulating hair growth. We have investigated the effects of a number of interleukins (IL). colony stimulating factors and tumour necrosis factors (TNF) on hair follicle growth in vitro. Dose-response studies showed that IL-1α. IL-1ß and TNF-o were potent inhibitors of hair follicle growth. The histology of hair follicles maintained with inhibitory doses of IL-1α. IL-1ß and TNF-α showed similar changes in hair follicle morphology, resulting in the formation of dystrophic anagen hair follicles. These changes in histology were characterized by the condensation and distortion of the dermal papilla, marked vacuolation of the hair follicle matrix, abnormal keratinization of the follicle bulb and inner root sheath, disruption of follicular melanocytes and the presence of melanin granules witbin the dermal papilla. Moreover, these changes in hair follicle morphology are similar to those reported in alopecia areata and suggest that IL-1α, IL-1ß and TNF-α may play an important part in the pathophysiology of inflammatory hair disease.     

A non-invasive tape absorption method for recovery of inflammatory mediators to differentiate normal from compromised scalp conditions

Background/aims: A simple non-invasive tape (Sebutape^TM) adsorption method was used to recover inflammatory proteins from normal and compromised human scalp (i.e. dandruff and seborrheic dermatitis) in order to assess the inflammatory and immunologic changes relevant to these clinical conditions. Methods: The scalps of subjects identified by a dermatologist as having either dandruff (n = 18), seborrheic dermatitis (n = 19) or normal scalp (n = 16) were visually graded to obtain total adherent scalp flaking scores (TASFS). Sebutape samples were then collected from both the high and low TASFS scalp sites using a one-minute tape application. To recover inflammatory molecules, tapes were extracted in buffered saline with sonication and the tape extracts analysed using commercial immunoassay methods for pro-inflammatory cytokines [i.e. interleukin-1α (IL-1α), IL-1β, IL-1 receptor antagonist (IL-1ra), IL-8 and tumor necrosis factor-α (TNF-α)] and the immunologic cytokines [i.e. IL-2, IL-4, IL-10, IL-12, IL-15 and interferon gamma (IFN-γ)]. Nitric oxide (NO) was also assayed on tape extracts using the Greiss reaction. To account for differences in protein loading on the tapes all cytokine and NO results were normalized using the total protein (TP) amounts recovered in tape extracts. Results: The IL-1α/TP levels recovered from dandruff and seborrheic scalps were significantly decreased (P = 0.03) compared to normal appearing scalp levels. The scalp levels of IL-1ra/TP and the ratio of IL-ra to IL-1α were significantly (P = 0.002) or directionally (P = 0.07) higher in seborrheic dermatitis scalps and dandruff scalps, respectively, compared to normal scalps. The IL-1ra and the IL-1ra/IL-1α ratio values correlated well with the TASFS. The TNF-α/TP levels recovered from dandruff scalps were significantly higher (P = 0.02) than levels recovered from seborrheic dermatitis and normal scalp subjects. IL-2/TP was significantly increased (P = 0.01) and IFN-γ and NO significantly decreased (P = 0.05) in the seborrheic dermatitis scalp samples compared to normal controls. Conclusion: The Sebutape method has proven useful for distinguishing normal from diseased scalp conditions. The cytokines recovered from the scalp tape samples showed distinct patterns that differentiated dandruff, seborrheic dermatitis and normal scalp populations. These methods may also prove useful for monitoring the clinical efficacy of therapeutic actives for treating dandruff and seborrhea.     

Epidermal stratification reduces the effects of UVB (but not UVA) on keratinocyte cytokine production and cytotoxicity

Ultraviolet (UV) radiation induces cytokine release from cultured keratinocytes as well as from epidermis in vivo. The purpose of this study was to determine whether differentiation of cultured keratinocytes into stratified epithelium decreases the effects of UVA and UVB radiation on cytokine release. Interleukin-1 (IL-1)α, IL-1β and tumor necrosis factor (TNF)-α release from human keratinocytes and reconstituted human epidermis was measured after exposure to UVA or UVB radiation. Release of IL-1α, IL-1β, and TNF-α was induced by both UVA and UVB radiation from both keratinocytes and reconstituted epidermis. Release of these cytokines was correlated with cytotoxicity. Keratinocyte cultures were far more sensitive to UVB radiation than reconstituted epidermis, in terms of both cytotoxicity and cytokine release. In contrast, epidermal stratification/differentiation had much less effect on the sensitivity to UVA radiation. We conclude that epidermal stratification and the formation of a stratum corneum provide protection against UVB radiation but have limited barrier effect against UVA radiation.     

Tumor necrosis factor-alpha blockade ameliorates inflammatory response in two children with chronic infantile neurological,cutaneous and articular syndrome

Chronic infantile neurological, cutaneous and articular (CINCA) syndrome is a rare autoinflammatory disease caused by monogenic defects in the NLRP3 gene. Pro-inflammatory cytokines such as interleukin (IL)-1β play a crucial role in the pathogenesis, and IL-1 receptor antagonists have been regarded as the mainstay therapy. Endogenous tumor necrosis factor (TNF)-α was found recently to be involved in the onset of the disease. Here, we report two Chinese children with CINCA syndrome who had elevated serum levels of TNF-α, with one carrying a novel mutation of c.1330T/G (p.444Phe/Val) in exon 3 of the NLRP3 gene. Anti-TNF-α (etanercept) therapy successfully alleviated both clinical symptoms and systemic inflammation after 6 months. These results suggest the complexity of the mechanisms of the disease and that TNF-α blockade will broaden the therapeutic approach for a subset of patients.     

磷酸二酯酶4抑制剂Hemay005对角质形成细胞炎性因子产生的影响

目的 为了探讨磷酸二酯酶4抑制剂Hemay005对体外培养的角质形成细胞株HaCaT细胞炎性因子产生的影响.方法 分别用重组人肿瘤坏死因子-α（rhTNF-α）、重组人干扰素-γ（rhIFN-γ）和312 nm窄波UVB诱导,采用MTT法检测Hemay005对HaCaT细胞增殖的影响,用ELISA法检测其对HaCaT细胞白细胞介素（IL）-8、sICAM-1和肿瘤坏死因子（TNF）-α产生的影响.结果 Hemay005对HaCaT细胞增殖及25 μg/L rhTNF-α诱导的IL-8产生无明显影响,但能剂量依赖性地抑制1 000 U/mL rhIFN-γ诱导的HaCaT细胞产生sICAM-1和124.2 mJ/cm2窄波UVB照射引起的HaCaT细胞产生TNF-α.结论 Hemay005可通过抑制角质形成细胞炎症因子的表达发挥抗炎作用.     

Evaluating the effects of sedentary behaviour on plantar skin health in people with diabetes

BackgroundDiabetes-Related Foot Ulcers (DRFUs) are a common and devastating consequence of Diabetes Mellitus and are associated with high morbidity, mortality, social and economic costs. Whilst peak plantar pressures during gait are implicated cited as a major contributory factor, DRFU occurrence has also been associated with increased periods of sedentary behaviour. The present study was designed aimed to assess the effects of sitting postures on plantar tissue health.MethodsAfter a period of acclimatisation, transcutaneous oxygen tensions (T_CPO₂) and inflammatory cytokines (IL-1α and IL-1RA) were measured at the dorsal and plantar aspects of the forefoot before, during and after a 20-min period of seated-weight-bearing in participants with diabetes (n = 11) and no diabetes (n = 10). Corresponding interface pressures at the plantar site were also measured.ResultsDuring weight-bearing, participants with diabetes showed increases in tissue ischaemia which were linearly correlated proportional to plantar pressures (Pearson's r = 0.81; p < 0.05). Within the healthy group, no such correlation was evident (p > 0.05). There were also significant increases in post seated weight-bearing values for ratio for IL-1α and IL-1RA, normalised to total protein, post seated weight-bearing in participants with diabetes compared to healthy controls.ConclusionThis study shows that prolonged sitting may be detrimental to plantar skin health. It highlights the need to further examine the effects of prolonged sitting in individuals, who may have a reduced tolerance to loading in the plantar skin and soft tissues.     

The effectiveness of a hydrocolloid crusting method versus standard care in the treatment of incontinence-associated dermatitis among adult patients in an acute care setting: A randomised controlled trial

IntroductionIncontinence-associated dermatitis (IAD) is a type of irritant contact dermatitis due to prolonged exposure of the skin to moisture induced by urine or/and faeces. The main principles when treating IAD involves protecting the skin from further exposure to irritants, establishing a healing environment, and eradicating skin infections. This study aimed to evaluate the effectiveness of the hydrocolloid crusting method (HCM) versus the standard care method (SCM) in treating IAD.MethodA randomised controlled trial was conducted in an acute tertiary hospital in Singapore between August 2019 to September 2021. Using computer-generated numbers, patients were randomised into either HCM or SCM treatment groups. HCM treatment involved cleansing the affected area with a pH-neutral non-rinse moisturising cleanser, and the application of alternate layers of hydrocolloid powder, and non-sting film barrier spray (repeated three times during each use). Patients in the SCM treatment group received the same cleanser followed by a 30% zinc oxide barrier cream. IAD was assessed daily for up to seven days by the wound care nurses using the IAD severity tool. The primary outcome of the study was the mean difference in IAD score per day between both methods.ResultsForty-four patients were eligible and recruited (22 in HCM; 22 in SCM). Patients in both groups were comparable in age and gender. IAD Category 2 was more predominant in both methods. The most common location of IAD was at the perianal skin and diarrhea related to gastroenteritis was the most prevalent cause of IAD. More patients in the SCM group (n = 12; 54.5%) had their IAD healed within seven days compared to HCM, (n = 7; 31.8%) group. However, the average decrease in IAD scores per day for both methods were found to be similar.ConclusionHCM can be considered as a treatment of IAD along with the use of SCM. A skin care regimen should include effective cleansing, skin protection, and moisturization in IAD management.     

Barrier disruption increases gene expression of cytokines and the 55 kD TNF receptor in murine skin

Abstract The signalling mechanisms that regulate epidermal permeability barrier homeostasis arc not known. Previous Northern blot analysis showed that both acute and chronic barrier disruption increase mRNA levels of several cytokines in murine epidermis. To further characterize the epidermal response to barrier abrogation, we used more sensitive, multi-probe RNasc protection assays to measure the mRNA levels of additional cytokines. as well as cytokine receptors in acute and chronic models of barrier disruption. Normal mouse epidermis expressed interleukin (IL)-lα, interferon-γ (IFN-γ). tumor necrosis factor-α (TNF-α) and IL-6 mRNAs. Following tape-stripping, only the mRNA levels for TNF-α, IL-lα, IL-lβ and IL-6 increased at 2.5 and 7 h, and returned toward normal levels by IX h. No mRNAs encoding TNF-β. IL-2, IL-3. IL-4 or IL-5, were detected in the epidermis either under basal conditions or after tape-stripping. Similarly, in a chronic model, essential fatty acid deficiency, epidermal levels of TNF-α, IL-lα, IL-lβ and IL-6 mRNAs, but not IFN-γ mRNA. were elevated over controls; and again, mRNAs for the remaining probed cytokines were not detected. In contrast, in the dermis. only IL-Iβ mRNA levels increased 2.5 h after tape-stripping, and remained elevated at I8 h. mRNAs encoding the IL-1 (p60), IFN-γ ami IL-6 receptors were present in epidermis, but their levels remained unchanged following either acute or chronic barrier disruption. In contrast, epidermal TNF (p55) receptor mRNA levels were increased by 87% (P<0.01) at 2.5 h, returned to control levels at 7 h and were increased by 68% (p<0.03) at 18 h after tape-stripping. The increase at 2 h was confirmed by Northern blot analysis and was not prevented by latex occlusion performed immediately after tape-stripping. mRNAs for the IL-I (p80) receptor and TNF (p75) receptor were not detected in epidermis. Low levels of TNF (p55) receptor mRNA were present in the dermis. and they remained unchanged after tape-stripping. The presence of specific receptor mRNAs in the epidermis and dermis suggests that these tissues are capable of responding in an autocrine and/or paracrine fashion to the cognate cytokines. These results suggest that epidermal cytokines produced after barrier disruption may initiate a cytokine cascade which could regulate cytokine and cytokine receptor production and/or inflammatory responses.