BACKGROUND: Interleukin-10(IL-10) was initially discovered and isolated on the basis of its ability to suppress cytokine by Thl helper T cell. Recently, the effect of IL-10 has been reported in cultured connective tissue cells. OBJECTIVE: To further investigate the mechanisms of IL-10 on extracellular matrix(ECM) homeostasis, we evaluated the expression of type I collagen and stromelysin-1 at the transcriptional level and also observed their promoters activities. METHODS: We examined that effect of the recombinant human IL-10 on the expression of genes involved in extracelluar matrix(ECM) synthesis and remodelimg in human dermal fibroblast cultures. Quantification of collagen synthesis, Northern blot analysis, transient transfection and a chloramphenicol acetyl transferase(CAT) assay were performed. RESULTS: In studying the dose and time response effect of IL-10 on collagen synthesis, maximal reduction was seen at 1.0 ng/ml concentration and 24 hours incubation. The synthesis of type I collagen mRNA was downregulated, while stromelysin-1 gene expression was enhanced by IL-10. In the transient transfection and CAT assay, the type I collagen promoter/CAT reporter gene construct showed downregulation by IL-10, while the stromelysin-1 gene promoter activities were upregulated. CONCLUSION: It is suggested that IL-10 differently regulates the transcriptional levels of type I collagen and stromelysin-1 gene and denoting IL-10 seems to take part in the homeostasis of ECM at pretranscriptional level.
Animals ; Blotting, Northern ; Cats ; Chloramphenicol ; Collagen ; Collagen Type I* ; Connective Tissue Cells ; Down-Regulation ; Fibroblasts* ; Gene Expression ; Genes, Reporter ; Homeostasis ; Humans ; Interleukin-10* ; RNA, Messenger ; Transfection

BACKGROUND: Recent clinial observations have suggested that colchicine, which is in frequent use in gout can affect the conneciive tissue metabolism in skin and other ti.ues. OBJECTIVE: This study suggest that further development of colchiine might provide a novel means of modulating collagen gen expression in patients with fibrotic disease. METHOD: We examed the effect of colchicine on the expres, on of collagen, fibronectin and collagenase gene by skin filroblast culture hy Nort.hern and dot-blot iybridization. Result : The rate of transcription of genomic DNA corresponding o type I collagen and libvonectin was reduced in colchicine-treated cultures but collagenase was not. reduced. Canclusion : The microti.ikile disruptive agent, colchicine, reduced the expression of type I collagen and fibronectin in a dose-(lependent manner. This st.udy suppor that colchicine is one of the promising antifibrogenic drugs curvently being tested as a treatment, of hun an fibrotic disease.
Colchicine* ; Collagen Type I ; Collagen* ; Collagenases ; DNA ; Fibroblasts ; Fibronectins ; Gene Expression* ; Gout ; Humans ; Metabolism ; Skin

BACKGROUND: bFGF, a member of the fibroblast growth factor family, potently induces vascular smooth muscle cell proliferation and decreased synthesis of the collagens. OBJECTIVE: For further investigation of the effect of bFGF on extracellular matrix homeostasis in the skin, we evaluated the expression of type I and type VII collagen gene at the transcriptional levels. METHOD: We examined that recombinant human bFGF affects the expression of genes involved in ECM synthesis and remodeling in human dermal fibroblasts cultures as judged by Northern blot analysis. RESULTS: The steady state levels of type I and VII collagen gene mRNA were decreased with age dependent pattern up to 0.13 and 0.44 folds respectively. The transcriptional levels of type I collagen mRNA were increased by TGF-B, treatment but markedly decreased by bFGF as well as TNF-a. But there were no synergistic effects bFGF and TNF-a on type I collagen gene expression. The levels of type VII collagen gene expression were increased by both bFGF and TGF-B,. The TNF-a showed slightly antagnostic effects on type VII collagen gene expression. CONCLUSION: The type I and VII collagen gene expression in dermal fibroblasts is clearly subjected to modulation by the cytokines including bFGF with uncoordinate regulatory pathway. In addition to its function of vascular proliferation, bFGF also may play a major role in physiologic skin condition and in repair process such as formation of a stable dermoepidermal junction during skin wound healing.
Blotting, Northern ; Cell Proliferation ; Collagen Type I ; Collagen Type VII ; Collagen* ; Cytokines ; Extracellular Matrix ; Fibroblast Growth Factors ; Fibroblasts* ; Gene Expression* ; Homeostasis ; Humans ; Methods ; Muscle, Smooth, Vascular ; RNA, Messenger ; Skin ; Wound Healing

BACKGROUND: Ultraviolet B(UVB) light, which can cause severe damage like induction and promotion of cancer, cutaneous inflammation and immunosuppression, represents one of the most important environmental impacts for humans. Keratinocytes are natural target cells of UVB in humans. NF-kB plays a role in the cell of the immune system, where it controls the expression of various cytokines and the major histocompatibility complex gene. OBJECTIVE: The purpose of this study was to examine the effect of UVB on the NF-kB and AP-1 activity in cultured human keratinocytes and fibroblasts. METHODS: Keratinocyte and fibroblast cultures were produced with DMEM medium. Cells were irradiated for 100J/m, 200J/m, 300 J/m. and Nuclear proteins were extracted. AP-1 and NF-kB activities were measured by the gel shift mobility assay. RESULTS: Gel shift mobility assay. 1. The NFkB activity was increased upon UVB in a dose dependent manner in the keratinocyte. 2. Enhanced levels of AP-1 binding activity in the radiated extracts frorn human skin keratinocytes were detected. 3. The levels of GRE (glucocorticoid responsive element) binding activity were similar in both radiated and unradiated extracts from fibroblasts and keratinocytes. CONCLUSIONS: The activities of NF-aB and AP-1 are increased following stimulation of a cell with UVB irradiation. Therefore, UV induced skin tumors, abnormal cell proliferation, cutaneous inflammation and immunosuppression, may be due to these transcriptional factors.
Cell Proliferation ; Cytokines ; Fibroblasts* ; Humans* ; Immune System ; Immunosuppression ; Inflammation ; Keratinocytes* ; Major Histocompatibility Complex ; NF-kappa B* ; Nuclear Proteins ; Skin ; Transcription Factor AP-1*

We report a case of primary cutaneous cryptococcosis on Rt. forehead and perioral area of 57 year old woman with non-insulin dependent diabetes mellitus and Lt. cerebral infarction. She had large ulcers with yellowish purulent exudates on Rt. forehead and perioral area for 2months. A histopathological examination from the lesion showed numerous encapsulated, round spores and the organisms were identified as Cryptococcus neoformans in a series of fun-gal studies. The patient received a 5-week course of IV and oral fluconazole with resolution of her skin lesion. The patient is free of any lesion several months after completing therapy. This experience supports the use of fluconazole as initial and single therapy in primary cutaneous cryptococcosis.
Cerebral Infarction ; Cryptococcosis* ; Cryptococcus neoformans ; Diabetes Mellitus ; Exudates and Transudates ; Female ; Fluconazole* ; Forehead ; Humans ; Skin ; Spores ; Ulcer

Lupus vulgaris is most prevalent on exposed parts, especially the face but can also develop on exetremities. Lupus vulgaris originates from tuberculosis elsewhere in the body by hematogenous, lymphatic, or contiguous spread. A 19-year-old male patient came to our department. The patient had had many recurrent oozing and verrucous plaques and crusts on the left foot for one year. A skin biopsy from the lesion on the left dorsum of the foot showed scattered well defined granulomas consisting of the epithelioid cell clusters with Langerhans and foreign body type giant cells in the mid dermis. Caseation necrosis was slight. There were no bacilli on AFB staining. The multi test CMI for tuberculin was highly positive. A chest X-ray did not show any abnormal findings. The presence of Mycobacterium tuberculosis DNA was demonstrated by polymerase chain reactions (PCR) for detection of mycobacterial DNA from a routinely prepared paraffin-embedded skin specimen. Herein we report a very atypical case of lupus vulgaris confirmed by PCR.
Biopsy ; Dermis ; DNA ; Epithelioid Cells ; Foot ; Giant Cells, Foreign-Body ; Granuloma ; Humans ; Lupus Vulgaris* ; Male ; Mycobacterium tuberculosis ; Necrosis ; Polymerase Chain Reaction ; Skin* ; Thorax ; Transplants* ; Tuberculin ; Tuberculosis ; Young Adult

BACKGROUND: Methods to detect and quanitify Mycobacterium leprae(M. leprae)are needed for studies involving the epidemiology, pathogenesis, and chemotherapy of leprosy. Serological assays and skin tests lack the sensitivity and specificity to serve as diagnostic tool for M. leprae infection. The polymerase chain reaction(PCR) based on the selective amplification of an 530-bp frangment of the gene encoding the proline-rich antigen of M. leprae was performed with sections of fixed or frozen biopsy samples from leprosy patients. OBJECTIVE: This study was done to investigate the applicability of PCR for the detection of low numbers of M. leprae in tissues and peripheral blood. METHODS: The PCR was used to amplify a 530-base-pair M. leprae DNA with the thermoxtable Taq DNA polymerase. RESULTS: The In frozen skin tissues and peripheral blood of leprosy patients. relatively high detection rates of PCR products was achieved by using direct gel analysis as well as Southern blot hybridization. CONCLUSION: These results suggest that PCR amplification for the detection of M. leprae may be useful for the epidemiologic study of large papulations as well as coinical astudies on the individual patients.
Biopsy ; Blotting, Southern ; DNA ; Drug Therapy ; Epidemiologic Studies ; Epidemiology ; Humans ; Leprosy ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Skin ; Skin Tests ; Taq Polymerase

BACKGROUND: It has become generally accepted that cigarette smoking contributes to accelerated coronary and peripheral vascular disease, pulmonary fibrosis and periodontal disease. Moreover, it has been postulated that cigarette smoking causes skin-aging. Many of cutaneous manifestations of nicotine which is a major component of the particulate phase of tobacco smoke are related to its vasoconstrictive and thrombotic effects on the peripheral vascular system. How-ever, direct effect of nicotine on extracellular matrix (ECM) proteins including collagens is not well established. OBJECTIVE: To evaluate the effect of nicotine on type I collagen gene expression in cultured human skin fibroblasts. METHODS: After exposure to different doses of nicotine on cultured human skin fibroblasts, we examined the expressions of α1(I) procollagen gene and fibronectin gene by Northern blot analysis and chloramphenicol acetyltransferase (CAT) assay with CAT construct containing the 3.5 kb COL1A2 promoter. RESULTS: In Northern blot hybridization, steady-state levels of α1(I) procollagen mRNA were decreased 0.8-fold at 1 µg/mL of nicotine, 0.5-fold at 10 µg/mL and 0.2-fold at 100 µg/mL, compared to untreated control. Those of fibronectin mRNA were decreased 0.9-fold, 0.7-fold, and 0.3-fold, respectively. In CAT assay, the relative COL1A2 CAT activity was 1.0 in the untreated control, 0.7 at a concentration of 1 µg/mL of nicotine, 0.5 at 10 µg/mL, and 0.3 at 100 µg/mL. CONCLUSION: These results indicate that nicotine is a down-regulator of collagen gene expression at transcriptional level in vitro. We speculate that nicotine may contribute to the skin-aging by modulation of extracellular matrix gene expression including collagen as well as by its vasoconstrictive and thrombotic effects.
Animals ; Blotting, Northern ; Cats ; Chloramphenicol O-Acetyltransferase ; Collagen ; Collagen Type I ; Extracellular Matrix ; Fibroblasts* ; Fibronectins ; Gene Expression ; Humans* ; Nicotine* ; Periodontal Diseases ; Peripheral Vascular Diseases ; Procollagen ; Pulmonary Fibrosis ; RNA, Messenger ; Skin* ; Smoke ; Smoking ; Tobacco