Keratinocytes-Derived Reactive Oxygen Species Play an Active Role to Induce Type 2 Inflammation of the Skin: A Pathogenic Role of Reactive Oxygen Species at the Early Phase of Atopic Dermatitis

Background@#Atopic dermatitis (AD) is characterized by chronic, relapsing skin inflammation (eczema) with itchy sensation. Keratinocytes, which are located at the outermost part of our body, are supposed to play important roles at the early phase of type 2 inflammation including AD pathogenesis. @*Objective@#The purpose of this study was to evaluate whether keratinocytes-derived reactive oxygen species (ROS) could be produced by the allergens or non-allergens, and the keratinocytes-derived ROS could modulate a set of biomarkers for type 2 inflammation of the skin. @*Methods@#Normal human epidermal keratinocytes (NHEKs) were treated with an allergen of house dust mites (HDM) or a non-allergen of compound 48/80 (C48/80). Then, biomarkers for type 2 inflammation of the skin including those for neurogenic inflammation were checked by reverse transcriptase-polymerase chain reaction and western immunoblot experiments. @*Results@#HDM or C48/80 was found to upregulate expression levels of our tested biomarkers, including type 2 T helper-driving pathway (KLK5, PAR2, and NF κ B), epithelial-cell-derived cytokines (thymic stromal lymphopoietin, interleukin [IL]-25, IL-33), and neurogenic inflammation (NGF, CGRP). The HDMor C-48/80-induced expression levels of the biomarkers could be blocked by an antioxidant treatment with 5 mM N-acetyl-cysteine. In contrast, pro-oxidant treatment with 1 mM H2O2 could upregulate expression levels of the tested biomarkers in NHEKs. @*Conclusion@#Our results reveal that keratinocytes-derived ROS, irrespective to their origins from allergens or non-allergens, have a potential to induce type 2 inflammation of AD skin.

CSP0510 Lotion as a Novel Moisturizer Containing Citric Acid and Trisodium Phosphate Relieves Objective and Subjective Symptoms of Atopic Dermatitis

BACKGROUND: Moisturizers with anti-inflammatory or anti-itch activity should be developed for the safe and effective management of atopic dermatitis (AD). OBJECTIVE: This study evaluated the efficacy of a newly developed moisturizer, CSP0510 lotion (Twolines Inc., Korea), containing citric acid (CA) and trisodium phosphate (TSP) as active ingredients, in mild to moderate AD. METHODS AND RESULTS: CSP0510 lotion applied twice daily for 4 weeks to eczematous lesions improved objective and subjective (itch) symptoms of AD. The physician's global assessment (PGA) score for objective symptoms decreased from 2.5±0.6 before application to 1.3±0.5 after application in the CSP0510-treated group (n=42, p<0.001). Also, the PGA score decreased from 2.3±0.6 to 1.9±0.5 by vehicle-treated (without CA and TSP) control group (p=0.001), but there was no statistical difference between CSP0510-treated and vehicle-treated groups (p=0.089). The visual analogue scale score for itch decreased from 4.8±1.3 to 2.0±0.9 in the CSP0510-treated group (p<0.001), and from 4.6±1.1 to 3.5±0.9 in the control group (p=0.075), showing a statistical significance between two groups (p=0.002). Our results in humans were further supported by in vitro and animal experiments. In HaCaT cells treated with compound 48/80 (7.5 µg/ml), CA:TSP (1:1, vol:vol) synergistically suppressed the compound 48/80-induced upregulation of thymic stromal lymphopoietin, nerve grow factor, and calcitonin gene-related peptide. Application of CSP0510 to the dorsal skin of hairless mice for 3 weeks suppressed the oxazolone-induced allergic skin inflammation. CONCLUSION: In conclusion, CSP0510 lotion has anti-itch and anti-inflammatory activity in the skin, which improves both objective and subjective symptoms of AD.

Ethanol Extract of Peanut Sprout Exhibits a Potent Anti-Inflammatory Activity in Both an Oxazolone-Induced Contact Dermatitis Mouse Model and Compound 48/80-Treated HaCaT Cells

BACKGROUND: We developed an ethanol extract of peanut sprouts (EPS), a peanut sprout-derived natural product, which contains a high level of trans-resveratrol (176.75 microg/ml) and was shown to have potent antioxidant activity. OBJECTIVE: We evaluated the potential anti-inflammatory activity of EPS by measuring its antioxidant potential in skin. METHODS: The anti-inflammatory activity of EPS was tested using two models of skin inflammation: oxazolone (OX)-induced contact dermatitis in mice and compound 48/80-treated HaCaT cells. As biomarkers of skin inflammation, cyclooxygenase-2 (COX-2) and nerve growth factor (NGF) levels were measured. RESULTS: OX-induced contact dermatitis was suppressed markedly in mice that were treated with an ointment containing 5% EPS as evidenced by a decrease in the extent of scaling and thickening (p<0.05) and supported by a histological study. COX-2 (messenger RNA [mRNA] and protein) and NGF (mRNA) levels, which were upregulated in the skin of OX-treated mice, were suppressed markedly in the skin of OX+EPS-treated mice. Consistent with this, compound 48/80-induced expression of COX-2 (mRNA and protein) and NGF (mRNA) in HaCaT cells were suppressed by EPS treatment in a dose-dependent manner. As an inhibitor of NF-kappaB, IkappaB protein levels were dose-dependently upregulated by EPS. Fluorescence-activated cell sorting (FACS) analysis revealed that EPS scavenged compound 48/80-induced reactive oxygen species (ROS) in HaCaT cells. CONCLUSION: EPS exerts a potent anti-inflammatory activity via its anti-oxidant activity in both mouse skin and compound 48/80-treated HaCaT cells in vitro. Compound 48/80-treated HaCaT cells are a useful new in vitro model of skin inflammation.

Effects of 630 nm Light-Emitting Diode Irradiation on Caveolin-1 and Procollagen I and III Expression in Human Dermal Fibroblasts / 대한피부과학회지

BACKGROUND: Recent studies indicate that light-emitting diodes (LED) may represent a novel and effective anti-aging light source for the skin. Among many candidate molecules known to control collagens, caveolin-1 (Cav-1) is known to play an inhibitory role in cutaneous collagen metabolism. OBJECTIVE: This study aimed to evaluate the effects of LED irradiation on the expression levels of Cav-1 and procollagens (proCOLs) in human dermal fibroblasts (HDFs). METHODS: Cultured HDFs were irradiated with 630 nm LED at different doses, and the mRNA and protein expression levels of Cav-1 and proCOLs I/III were analyzed. RESULTS: In LED-irradiated HDFs, mRNA and protein levels of Cav-1 were found to be down-regulated, whereas those of proCOLs I/III were up-regulated in a dose-dependent manner. A negative correlation between Cav-1 and proCOLs was verified in Cav-1 siRNA transfected HDFs. LED was moreover found to result in up-regulation of transforming growth factor (TGF)-beta1 and its receptors (TbetaRI, TbetaRII), SMAD1, and SMAD2 mRNA levels, indicating that LED may activate the TGF-1/TbetaR/SMAD pathway in HDFs. CONCLUSION: The anti-aging effects of 630 nm LED on human skin are likely mediated by up-regulation of proCOLs I/III and inhibition of Cav-1 expression levels in HDFs.

Up-Regulation of Cyclooxygenase 2 and Matrix Metalloproteinases-2 and -9 in Cutaneous Squamous Cell Carcinoma: Active Role of Inflammation and Tissue Remodeling in Carcinogenesis

BACKGROUND: Tissue inflammation and remodeling have been extensively studied in various tumors in relation with their invasiveness and metastasis. OBJECTIVE: The purpose of this study was to investigate the change in tissue inflammation and remodeling markers in cutaneous squamous cell carcinoma (SCC). METHODS: Expression levels of cyclooxygenase-2 (COX-2) as an inflammatory marker and matrix metalloproteinases-2 and -9 (MMPs 2/9) as remodeling markers were studied in mouse and human SCCs. Western blot analysis and RT-PCR for COX-2 and MMPs 2/9 were performed with skin samples from SCC patients and chronic ultraviolet B (UVB)-induced SCC from hairless mice. RESULTS: mRNA and protein levels of COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in mouse SCCs, which were induced by chronic UVB irradiation. Consistently, COX-2 and MMPs 2/9 were up-regulated with the higher sensitivity for MMP-9 in human SCCs. CONCLUSION: COX-2 and MMPs 2/9 are up-regulated in well-differentiated cutanous SCC. Our findings indicate that inflammatory and tissue remodeling processes are actively induced during carcinogenesis of cutaneous SCC.

Expression of Peroxiredoxin I in the Epidermis of Vitiligo / 대한피부과학회지

BACKGROUND: Although the pathogenesis of vitiligo isn't fully understood, a recent study demonstrates that oxidative stress plays an important role to induce vitiligo. Peroxiredoxin (Prx) is a novel peroxidase family to remove hydrogen peroxide using thioredoxin system, which is consisted of thioredoxin, thioredoxin reductase, and NADPH. OBJECTIVE: This study aimed to investigate the change of expression of Prx I to elucidate the role of oxidative stress in the pathogenesis of vitiligo. METHODS: Sample specimens were obtained from the lesional skin of vitiligo patients, and non-depigmented skin was obtained from the perilesional area as control samples. The skin samples were immediately frozen using liquid nitrogen, and then section samples were prepared to perform immunohistochemical staining with antibodies for Prx I. Some of the skin biopsy samples were used for primary culture of keratinocytes. Protein extracts from the expanded keratinocytes were prepared for Western blot analysis of Prx I. RESULTS: In vitiligo, the ubiquitous expression of Prx I in all layers of epidermis, which was also observed in the normal perilesional skin, was reduced in the depigmented lesion of vitiligo patients. The reduction of Prx I was remarkable from the lesions which were exposed to sunlight. Consistently, Prx I expression from the lesional keratinocytes were noticeably reduced in comparison with that from perilesional keratinocytes. CONCLUSION: Our results showing that Prx I is impaired in the epidermis of depigmented lesions of vitiligo patients suggest that oxidative stress is an important factor to induce vitiligo.

Modulation of Peroxiredoxin I Expression by UVB Irradiation in Human Keratinocytes: H2O2-mediated Modulation of Peroxiredoxin I / 대한피부과학회지

BACKGROUND: Peroxiredoxin I (Prx I) is part of an oxidative stress defense system with thioredoxin peroxidase activity to eliminate hydrogen peroxide (H2O2). UV irradiation is one of the major sources to produce H2O2, which should then be scavenged by antioxidant systems to maintain functional integrity of the skin. OBJECTIVE: This study aimed to evaluate the modulation of Prx I by ultraviolet B (UVB) irradiation in human epidermal keratinocytes. The modulation of Prx I expression by H2O2 was also evaluated. METHOD: Primary culture of epidermal keratinocytes was performed, and sub-confluent cells were irradiated with UVB irradiation (20mJ/cm(2)). Western blot and Northern blot analysis were performed after the cells were harvested at different time-points after UVB irradiation. Prx I expression and intracellular levels of H2O2 were evaluated in the cells which had been irradiated with different doses of UVB. The localization of Prx I expression was identified by immunocytochemical staining. RESULTS: UVB irradiation induced Prx I mRNA and protein expressions from 3 h and 6 h after irradiation, respectively, indicating that UVB induced Prx I expression at a transcription level. Intracellular H2O2 levels were steadily increased as keratinocytes were irradiated with increasing doses of UVB. Next, when keratinocytes were treated with 0.1-10.0mM of H2O2, the marked induction of Prx I protein expression was observed above 1 mM H2O2 at a time-dependent manner (after 6 h). The H2O2-induced Prx I expression was abolished by N-acetyl-L-cysteine, a H2O2 scavenger, pre-treatment. In 2D-gel electrophoresis, the active reduced form of Prx I was rapidly transformed into the oxidized, inactive form, and then it restored to the reduced form by H2O2 treatment, suggesting that Prx I was active in responding to the H2O2-induced oxidative stress. CONCLUSION: UVB irradiation up-regulates Prx I by the mediation of H2O2 in the keratinocytes.

Serum Stem Cell Factor (SCF) Levels and In Vitro Response of Hematopoietic Progenitors to Recombinant Human SCF (rhSCF) in Patients with Aplastic Anemia / 대한혈액학회지

BACKGROUND: Stem cell factor (SCF), which is produced from stromal cells of bone marrow, is a growth factor acting in early stages of hematopoiesis. Bone marrow failure in aplastic anemia seems to be mainly due to hematopoietic stem cell defect. However, there has been some evidence that microenvironmental defects contribute to the pathophysiology of this disorder. METHODS: We measured serum SCF levels in 29 patients with aplastic anemia at diagnosis using ELISA to define the status of soluble SCF in this disorder. We also examined the effect of serum from 15 patients with aplastic anemia on the colony growth (CFU-GM and BFU-E) of normal bone marrow cells and the ability of recombinant human SCF to stimulate in vitro colony growth of bone marrow cells from 10 patients with aplastic anemia. RESULTS: Levels of serum SCF in patients with aplastic anemia were not different from those in 25 healthy controls (1,493+/-392pg/mL vs. 1,563+/-322pg/mL) or those in patients with idiopathic thrombocytopenic purpura, acute myeloid leukemia, and myelodysplastic syndrome. No correlation was found between serum SCF levels and hematologic parameters at diagnosis such as neutrophil count, hemoglobin, and platelet count. No difference was noted in the serum SCF levels according to severity of the disease. The effect of serum from patients with aplastic anemia on the colony growth of normal bone marrow cells were variable and were not correlated with serum SCF levels. rhSCF increased the colony numbers of bone marrow cells from patients with aplastic anemia. CONCLUSION: These results indicate that soluble SCF is not deficient in aplastic anemia, but SCF can stimulate the proliferation of bone marrow cells from aplastic anemia.

Effect of FCL (5-FU, carboplatin, leucovorin) Chemotherapy in Advanced Head and Neck Cancer / 대한암학회지

PURPOSE: The purpose of this study was to evaluate the efficacy and the toxicities of recently developed second generation platinum, carboplatin in combination with 5-fluorouracil and leucovorin in head and neck cancer patients. PATIENTS AND METHODS: Between March 1993 and Apirl 1996, 22 patients with locally advanced/metastatic head and neck cancer were treated with FCL combination chemotherapy. Of these 20 patients were evaluable. RESULTS: Median age was 58 years. The number of patients with stage III and IV patients was 4 and 18 respectively. Among the 20 evaluable patients, 1 (7.2%) achieved a complete response and 8 (40%) achieved partial responses. The median duration of the response was 24 weeks and the median survival duration was 12 months. Out of 77 chemotherapy cycles, 1 patient (1.3%) had anemia of WHO grade 2, 7 patients (9.1%) experienced leukopenia (WHO grade > or =2) and 7 (9.1%) experienced thrombocytopenia (WHO grade > or =2). Non-hematologic toxicities were mild; nausea and voming of WHO grade > or =2 was 12 (15.6%), stomatitis (WHO grade > or =2) was 6 (7.8%). CONCLUSION: FCL chemotherapy was effective in locally advanced head and neck cancer. Toxocities were minimal compaired to cisplatin based combination chemotherapy.Further studies on increased dose of FCL chemotherapy in head and neck cancer patients is warranted.

The Effect of Dehydroepiandrosterone on Inhibition of Carcinogenesis and Induction of Apoptosis in Murine Hepatoma Model

Tumor suppressive effect of dehydroepiandrosterone (DHEA) on the experimentally induced hepatocellular carcinoma was investigated, especially focusing on glutatione transferase and transglutaminase with aptosis in the carcinogenesis. The chemical hepatocarcinogenic procedure of Solt-Farber method was used on Sprague-Dawley rats. Experimental groups were divided into AA group treated by the standard Solt-Farber regimen of diethylnitrosamine (DEN) and 2-acetamidofluorene (AAF) and AD group treated with DHEA simultaneously with AAF and the AAD group treated by DHEA after treatment with AAF. Each group was divided by time sequence further into four subgroups, GI (8wk), G2 (16wk), G3 (28wk), and G4 (36wk). For neoplastic lesion, the immuno histochemical study with anti GSTP antibody was carried out, while the activity and expression of TGase was compared at the same time. The results were summarized as follows; GST-P positive foci detected in AD groups were significantly more suppressed by DHEA treatment than AA groups (P<0.05). AD groups. AD group showed higher activities of TGase than AA groups (P<0.05), which was confirmed by Western and Northern blot analysis. But the number of apoptotic bodies was not correlated with activity and expression of TGase in the nodule. These results suggest that the suppressive effect of DHEA on the murine hepatocellular carcinogenesis might be operating on the promotion process of carcinogenesis rather than regression process of transformed hyperplastic nodules.

Suppression of chronic myelogenous leukemia colony growth and K562 cell proliferation by interleukin-1 receptor antagonist / 대한혈액학회지

No abstract available.