We investigated the question of whether cholesterol catabolite can influence expression of inflammatory cytokines via Toll-like receptors (TLR) in monocytic cells. Treatment of THP-1 monocytic cells with 27-hydroxycholesterol (27OHChol) resulted in induction of gene transcription of TLR6 and elevated level of cell surface TLR6. Addition of FSL-1, a TLR6 agonist, to 27OHChol-treated cells resulted in transcription of the IL-1alpha gene and enhanced secretion of the corresponding gene product. However, cholesterol did not affect TLR6 expression, and addition of FSL-1 to cholesterol-treated cells did not induce expression of IL-1alpha . Using pharmacological inhibitors, we investigated molecular mechanisms underlying the expression of TLR6 and IL-1alpha. Treatment with Akt inhibitor IV or U0126 resulted in significantly attenuated expression of TLR6 and IL-1alpha induced by 27OHChol and 27OHChol plus FSL-1, respectively. In addition, treatment with LY294002, SB202190, or SP600125 resulted in significantly attenuated secretion of IL-1alpha . These results indicate that 27OHChol can induce inflammation by augmentation of TLR6-mediated production of IL-1alpha in monocytic cells via multiple signaling pathways.
Cholesterol ; Cytokines ; Inflammation ; Interleukin-1 ; Interleukin-1alpha* ; Toll-Like Receptors*

BACKGROUND: Keratinocyte-derived interleukin-1(IL-1)alpha is one of the key cytokines in initiation of cutaneous inflammation. Release of IL-1alpha from human keratinocytes may be induced by proinflammatory stimuli including ultraviolet B(UVB) irradiation, and subsequently, keratinocyte-derived IL-1alpha may exert numerous paracrine and autocrine effects. 1,25-dihydroxyvitamin D3(1,25(OH)2D3) is involved in the regulation of keratinocyte proliferation and differentiation and is also recognized to have immunoregulatory properties such as an antiinflammatory effect. OBJECTIVE: The purpose of this study was to investigate the in vitro effects of 1,25-(OH)2D3 on the production of IL-1alpha by UVB irradiation in cultured human keratinocyte cell line HaCaT cells. RESULTS: are summerized as follows; 1. The vialility of cultured HaCaT cells measured by MTS assay at 24 hours after UVB irradiation was significantly reduced at the doses of above 100 mJ/cm2 of UVB(p<0.05). 2. The secretion of IL-1alpha by HaCaT cells was significantly increased at the doses of above 30 mJ/cm2 of UVB(p<0.05). UVB irradiation could not influence on the secretion of IL-1beta by HaCaT cells. 3. At the concentrations of 10-8M and 10-6M of 1,25(OH)2D3, the production of IL-1alpha by HaCaT cells(48 hours after 100 mJ/cm2 UVB irradiation) was significantly inhibited in both culture supernatants and cell lysates(p<0.05). CONCLUSION: UVB irradiation increased the production of IL-1alpha by HaCaT cells and this stimulatory effect on the production of IL-1alpha induced by UVB irradiation was suppressed by 1,25-(OH)2D3. Calcipotriol(MC-903) had similar suppressive effect on the production of IL-1alpha induced by UVB irradiation in HaCaT cells to that of 1,25(OH)2D3.
Calcitriol* ; Cell Line* ; Cytokines ; Humans* ; Inflammation ; Interleukin-1alpha* ; Keratinocytes*

BACKGROUND: Lipid peroxide (LPO) in comedones, which are produced as a result of sebum oxidation, might potentially induce interleukin-1alpha (IL-1alpha) and exacerbate comedogenesis and inflammatory changes in comedones. OBJECTIVE: To investigate the relationship of proinflammatory cytokines and LPO levels in the extracts of comedones with the acne of clinical difference between smokers and non-smokers, and with the severity and distribution of the acne lesions. METHODS: Twenty-two non-smoking and 21 smoking adult acne patients were evaluated by comedone extraction and measurement of proinflammatory cytokines and LPO levels. Acne severity and distribution of the lesions were also analyzed. RESULTS: Relative to the non-smoking group, smokers had significantly higher levels of IL-1alpha and LPO in comedones. Their levels showed a positive correlation. However, there were no statistically significant difference between the severity or distribution of the disease and the levels of LPO and IL-1alpha in comedones. CONCLUSION: Smoking may be involved in the pathogenesis of adult acne by increasing the oxidative stress that results in subsequent accumulation of LPO in comedones.
Acne Vulgaris* ; Adult* ; Cytokines ; Humans ; Interleukin-1* ; Interleukin-1alpha* ; Oxidative Stress ; Sebum ; Smoke ; Smoking ; Tobacco Products*

PURPOSE: This study was to identified differentiated genes concerned with apoptosis that are up-regulated or down-regulated in OLETF corneal keratocytes stimulated with interleukin-1alpha. METHODS: Otsuka Long Evans Tokushima Fatty (OLETF) corneal keratocytes were primarily cultured and treated with 20 ng/ml of IL-1a for 6 hours. Hybridization was carried out using Oligonucleotide microarray. A sample of normal keratocytes was labeled with Cy 3, and a diabetic keratocytes sample was labeled with Cy 5. RESULTS Diabetes showed 20 down-regulated genes and 24 up-regulated genes compared with normal keratocytes. Also, IL-1alpha-treated diabetic keratocytes expressed 31 down-regulated and 33 up-regulated genes. Compared with diabetic keratocytes, the newly expressed genes in OLETF treated with IL-1alpha were Bcl, Lam, Timp, Mmp, Socs, Bmp, Ccl, Lcn, C7, etc. CONCLUSIONS: There were some genes related with apoptosis that expressed differently between normal and diabetic keratocytes of OLETF. Also, IL-1alpha may stimulate differing effects of apoptosis on diabetic corneal keratocytes. Studies analyzing the apoptotic significance of these differences identified in diabetic keratocytes are needed.
Animals ; Apoptosis* ; Corneal Keratocytes ; Diabetes Mellitus ; Interleukin-1alpha ; Oligonucleotide Array Sequence Analysis ; Rats

PURPOSE: This study was to identified differentiated genes concerned with apoptosis that are up-regulated or down-regulated in OLETF corneal keratocytes stimulated with interleukin-1alpha. METHODS: Otsuka Long Evans Tokushima Fatty (OLETF) corneal keratocytes were primarily cultured and treated with 20 ng/ml of IL-1a for 6 hours. Hybridization was carried out using Oligonucleotide microarray. A sample of normal keratocytes was labeled with Cy 3, and a diabetic keratocytes sample was labeled with Cy 5. RESULTS Diabetes showed 20 down-regulated genes and 24 up-regulated genes compared with normal keratocytes. Also, IL-1alpha-treated diabetic keratocytes expressed 31 down-regulated and 33 up-regulated genes. Compared with diabetic keratocytes, the newly expressed genes in OLETF treated with IL-1alpha were Bcl, Lam, Timp, Mmp, Socs, Bmp, Ccl, Lcn, C7, etc. CONCLUSIONS: There were some genes related with apoptosis that expressed differently between normal and diabetic keratocytes of OLETF. Also, IL-1alpha may stimulate differing effects of apoptosis on diabetic corneal keratocytes. Studies analyzing the apoptotic significance of these differences identified in diabetic keratocytes are needed.
Animals ; Apoptosis* ; Corneal Keratocytes ; Diabetes Mellitus ; Interleukin-1alpha ; Oligonucleotide Array Sequence Analysis ; Rats

It was investigated if regulation of the trabecular extracellular matrix tumover rate and remodeling plays an important role in decreasing outflow resistance by determining the effect of intracamerally given interleukin-1alpha a known stimulator of the expression of trabecular matrix metallopro-teinases, on outflow facility of albino rat eyes. Forty normal albino rats (Sprague dawley) weighing 250 to 300gm were studied. Rats were anesthetized by intraperitoneal pentobarbital sodium(30mg/kg) injection. The rats were grouped into 4 groups and given 5, 10, 25, 50 units of interleukin-1alpha injected intracamerally in one eye of each rat. Bovine serum albumin in phosphate buffered saline, which was used to dissolve the interleukin-1alpha, was injected in the other eye as a control. Outflow facility was measured by two level constant pressure perfusion at 1, 3, and 7 days after injection. The eyes treated with 50 units of interleukin-1alpha showed a statistically significant increase of outflow facility by 37% compared to the contralateral control eyes at 3 days after injection, but retumed to normal level in 7 days. The eyes treated with 5, 10, 25, 50 units of interleukin-1alpha increased the outflow facility, supporting the hypothesis that regulation of trabecular meshwork extracellular matrix plays a role in trabecular outflow resistance.
Animals ; Extracellular Matrix ; Interleukin-1alpha* ; Interleukins ; Pentobarbital ; Perfusion ; Rats* ; Serum Albumin, Bovine ; Trabecular Meshwork

PURPOSE: Immature immunological defens mechanism in the neonate may contribute to the high susceptibility to overwhelming sepsis. S. aureus TSST-1 and E. coli LPS known as one of the important pathogens of septic shock or toxic shock induce massive release of various cytokines, such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-1alpha (IL-1alpha) produced from peripheral blood mononuclear cells (PBMC). In contrast, limited information has been provided so far concerning the capacity of cytokine production from neonatal immune cells. METHODS: This study was conducted to compare the secretion of proinflammatory cytokines, such as IL-l and TNF-alpha from cord blood PBMC to those from adult blood PBMC stimulated by S. aureus TSST-1 and E. coli LPS. RESULTS: 1) IL 1-alpha was secreted in a time-dependent manner from cord & adult blood PBMC stimulated with several cytokine inducers, and LPS stimulated adult & cord blood PBMC secreted IL 1-alpha in a dose-dependent manner. 2) TNF-alpha secretion from cord blood PBMC stimulated with LPS and IFN- significantly decreased in a time dependent manner, but not from adult PBMC. And secretion of TNF- from cord blood PBMC reached the highest level 24 hours after stimulated with LPS or IFN-gamma. The secretion of TNF-alpha from adult blood PBMC showed similar pattern to those from cord blood PBMC, but higher than cord blood PBMC. 3) IL-1alpha & TNF-alpha secretion from cord & adult blood PBMC stimulated with TSST-1 had no significant difference except in TNF- secretion by TSST-1 at 96 hours. 4) The secretion of IL-1alpha from adult PBMC stimulated with LPS showed higher and longer than that from cord blood PBMC. 5) IL-1alpha & TNF-alpha secretion from cord & adult blood PBMC stimulated with IFN-gamma had no significant difference. CONCLUSIONS: In summary, proinflammatory cytokines such as IL-1alpha, TNF-alpha in cord blood PBMC were secreted in a time dependent manner, but the amounts of IL-1alpha and TNF-alpha secretion were lesser than those of adult blood PBMC, especially stimulated by LPS. These results suggest that increased susceptibility to infection in neonatal period may be partially from a functional immaturity of cord blood mononuclear cells.
Adult* ; Cytokines ; Fetal Blood ; Humans* ; Infant, Newborn ; Interleukin-1alpha* ; Sepsis ; Shock, Septic ; Tumor Necrosis Factor-alpha*

BACKGROUND: Hydroquinone (HQ) is frequently combined with retinoic acid (RA) to enhance lightening efficacy, which may also affect skin irritancy. Although skin irritation leads to postinflammatory hyperpigmentation, little research has been performed to compare skin irritancy between each component and the combination. OBJECTIVE: This study was done to examine whether HQ-RA combination increased skin irritation induced by HQ or RA alone. METHODS: Patch testing was performed using maximum therapeutic and higher concentrations of HQ and RA in 10 volunteers, and then, it was performed using their popular therapeutic concentrations and combination in the other 20 volunteers. In vitro irritation was also assessed in primary cultured normal human keratinocytes treated with 80% and 50% cell survival doses of HQ, 80% cell survival dose of RA, and their combination. RESULTS: The combination in patch testing induced stronger erythema than the corresponding concentrations of HQ and RA, which was remarkable with use of combination of higher concentrations. In cultured keratinocytes, the RA combination significantly decreased cell viability, but increased cytotoxicity and extracellular interleukin 1 alpha release with corresponding doses of HQ. CONCLUSION: The results of patch tests and in vitro irritation assessment tests suggested that HQ and RA increased skin irritation when used in combination.
Cell Survival ; Erythema ; Humans ; Hyperpigmentation ; In Vitro Techniques ; Interleukin-1alpha ; Keratinocytes ; Patch Tests ; Skin* ; Tretinoin ; Volunteers

BACKGROUND: The Fas antigen is a cell surface molecule that mediates apoptosis in many cell types. Matsues group indicated that keratinocytes constitutively express the Fas antigen and apoptosis was induced only on pretreatment with interferon-r (IFN-y) in cultured normal human keratinocytes (NHK). OBJECTIVE: We undertook this study to determine the induction of apoptosis by Fas antibody alone and/or in combination with IFN y, IL-1a in normal human keratinocytes (NHK) and transitional epithelioma cell lines (KB cell) which had lower levels of intracellular IL-1 receptor antago- nists (IL-1ra ). METHODS: We used cultured NHK and KB cells. Each cell was treated with IFN-r, IL-la and Fas antibody for induction of apoptosis. For quantifying the apoptosis, index fluorescent DNA- binding dyes were used. Result: Fas antibody alone could induce apoptosis not only in KB cells but also in NHK cells. The combination of Fas antibody and IFN-r enhanced the induction of apoptosis in NHK and KB cells. The IL-la alone could induce apoptosis only in KB cells which had relatively small amounts of IL-1ra compared to NHK. CONCLUSION: Our result may indicate that Fas antigen in human keratinocytes can regulate normal epidermal cellular differentiation and proliferation.
Antigens, CD95 ; Apoptosis* ; Carcinoma ; Cell Line ; Coloring Agents ; Humans* ; Interleukin 1 Receptor Antagonist Protein ; Interleukin-1 ; Interleukin-1alpha ; KB Cells* ; Keratinocytes*

BACKGROUND: One of the limitation during the irradiation of malignant tumor is hazard to normal tissue although it is important and effective tool for treating malignant tumor. We studied the role of interleukin-1 alpha (IL-1alpha) and interleukin-6 (IL-6) in the radiation-induced lung injury especially on fibrosis. METHODS: We irradiated right-side lungs of thirty Sprague-Dawley rats with single fraction of 20 Gy and then sacrificed the animals until 20th week at intervals of two weeks. Both irradiated and unirradiated lung tissues were stained hematoxilin and eosin, Masson trichrome, reticulin and immunohistochemical staining for IL-1alpha and IL-6. The degree of the staining for IL-1alpha and IL-6 were examined semiquantitatively. RESULTS: Two weeks after irradiation interstitial edema and capillary congestion appeared, followed by increase of the monocytes infiltration and proteinaceous material during 4th and 8th week. After eight weeks of irradiation, collagen and reticulin fibers were detected along alveolar wall. 12th to 20th week, fibrosis in interstitium, decreased number of alveoli and thickening of bronchial wall were observed. The degree of immunohistochemical staining for IL-1alpha and IL-6 was increased rapidly during the first three week and then decreased slowly, but remain incresed until 20th week. CONCLUSION: Our Study demonstrate the early and persistent elevation of cytokines IL-1alpha and IL-6 by immunohistochemical stain in rat lung following pulmonary irradiation. We think cytokines are produced immediately after irradiation, make collagen genes turn on and perisist until the expression of late effects become apparent pathologically and clinically.
Animals ; Capillaries ; Collagen ; Cytokines ; Edema ; Eosine Yellowish-(YS) ; Estrogens, Conjugated (USP) ; Fibrosis ; Interleukin-1* ; Interleukin-1alpha ; Interleukin-6* ; Lung Injury ; Lung* ; Monocytes ; Rats* ; Rats, Sprague-Dawley ; Reticulin