ㄡㄡ莢ざ呿蒂菜⻆扡粗喀莘鍚莘’噤狤

ㄡㄡ莢ざ呿蒂菜⻆扡粗喀莘鍚莘’噤狤

-42- Matrix(2) 9/4 Room-C
NITRIC OXIDE INDUCES PROTEASES AND COLLAGEN PRODUCTION IN DERMAL FIBROBLASTS.
kentaro Omori, Masashi Ono, Hiroaki Ueki.
Department of Dermatology, Kawasaki Medical School, Kurashiki, Japan.
Nitric Oxide (NO), a short-lived radical, acts in vasorelaxsation and neurotransmission. On the other hand, the production of NO in tissue inflammation was evidenced by induction of NO synthease (NOS) isoformes in infiltrated cells such as macrophages and fibroblasts. The role of NO in inflammation has not been well-understood. NO could have a variety of functions, which is a radical and might have direct toxicity in tissue, or acts as a signal transmitter and elicits cellular responses. It has also been controversial whether effects of NO in inflammation is "proinflammatory" or "anti-inflammatory". In the present study, normal human fibroblasts were cultured with NO generators (SNP, SNAP or DETA-NONOate). Effects of NO in fibroblasts were monitored by cell proliferation, methionine incorporation, expression of matrixes and proteases. In results, NO induced proteases including collagenase below the toxic concentration. It also induced collagen 炒 and 炕 production. Our observation may evoke an important role of NO for signal transmission in inflammation and wound healing.

-43- Matrix(2) 9/4 Room-C
TNF-a INCREASES TYPE I AND III COLLAGEN GENE EXPRESSIONS IN SELF-PRODUCED THREE-DIMENSIONAL FIBROBLASTS CULTURES.
Madoka. Sato, Osamu Ishikawa, Yoshiki Miyachi.
Department of Dermatology, Gunma University School of Medicine, Maebashi, Gunma, Japan.
We have recently established a novel fibroblast culture system supplemented with L-ascorbic acid 2-phosphate. The addition of L-ascorbic acid 2-phosphate could render human dermal fibroblasts to organize a dermis like structure by accumulating collagens and extracellular matrices. The purpose of this study is to examine the effects of TNF-a on collagen gene expressions by human dermal fibroblasts in this culture system in comparison with those in monolayer culture. TNF-a suppressed the expressions of proa1(I) and proa1(III) collagen mRNA in monolayer culture. In contrast, these expressions were elevated in three-dimensional culture system. TNF-a increased the mRNA expressions of collagenase and TIMP-1 in both culture systems. These data suggest that responses of human dermal fibroblasts to TNF-a is distinct under the different culture conditions. Since our three dimensional culture system is quite similar to the dermis both morphologically and biochemically, TNF-a possibly regulates both collagen production and destruction in vivo, if this three-dimensional culture system really reflects the HDFs' nature in human dermis.

-44- Matrix(2) 9/4 Room-C
THE EFFECTS OF INTERLEUKIN-1b AND INTERLEUKIN -10 ON THE ELASTIN GENE EXPRESSION.
Bon-Sik Koo, Soo-Deuk Kong , Byung - Chun K i m, Kyu-Suk L e e, Joon-Young Song.
Department of Dermatology, Keimying University Medical School, Taegu, Korea.
T h e e l a s t i c f i b e r s a r e a i m p o r t a n t f i b r i l l a r c o m p o n e n t o f t h e e x t r a c e l l u l a r m a t r i x o f s k i n p r o v i d i n g e l a s t i c i t y a n d r e s i l i e n c e t o t h e s k i n . I n p h o t o a g e d s k i n , a c c u m u l a t i o n o f a b n o r m a l e l a s t i c f i b e r s i s a c h a r a c t e r i s t i c h i s t o l o g i c f i n d i n g . R e c e n t m o l e c u l a r b i o l o g i c a l s t u d i e s h a v e r e v e a l e d t h a t u l t r a v i o l e t ( U V ) i r r a d i a t i o n a c t i v a t e d e l a s t i n g e n e e x p r e s s i o n i n v i v o a n d i n v i t r o . F o l l o w i n g U V i r r a d i a t i o n , e p i d e r m a l k e r a t i n o c y t e s p r o d u c e m a n y c y t o k i n e s s u c h a s i n t e r l e u k i n ( I L ) - 1 , t u m o r n e c r o s i s f a c t o r - ( a n d I L - 1 0 . S o , U V i n d u c i b l e c y t o k i n e s e l a b o r a t e d b y k e r a t i n o c y t e s a n d i n f l a m m a t o r y c e l l s m a y p l a y a r o l e i n p a t h o g e n e s i s o f d e r m a l c h a n g e i n p h o t o a g i n g . I n t h i s s t u d y , w e h a v e e x a m i n e d t h e e f f e c t s o f I L - 1 a n d I L - 1 0 o n e l a s t i n g e n e e x p r e s s i o n i n h u m a n s k i n f i b r o b l a s t s i n c u l t u r e b y N o r t h e r n h y b r i d i z a t i o n , t r a n s i e n t t r a n s f e c t i o n a n d i n d i r e c t i m m u n o f l u o l e s c e n c e e x p e r i m e n t s . T h e r e s u l t s i n d i c a t e u p - r e g u l a t i o n o f e l a s t i n g e n e e x p r e s s i o n a t t h e t r a n s c r i p t i o n a l l e v e l , I L - 1 a n d I L - 1 0 e l a b o r a t e d b y m a n y c e l l s s u c h a s k e r a t i n o c y t e s , i n f l a m m a t o r y c e l l s f o l l o w i n g U V i r r a d i a t i o n p l a y a r o l e , a t l e a s t i n p a r t , i n p a t h o g e n e s i s o f p h o t o a g i n g .

-45- Matrix(2) 9/4 Room-C
THE EFFECT OF ELECTRICAL ACUPUNCTURE STIMULATION USING THERMOGRAPHY AND PLASMA ENDOTHELIN(ET-1), SEROTONIN AND NO LEVELS IN PATIENTS WITH SYSTEMIC SCLERODERMA(PSS).
Manabu Maeda, Hisaya Kachi, Naoki Ichihashi, Yasuo Kitajima.
Department of Dermatology, Gifu University, School of Medicine, Gifu, Japan.
Ten patients with PSS(Barnett type I;2, II;5, III;3 cases), 6 cases of SSSD(atypical or incomplete type of PSS accompanied with possible Sj蚧ren syndrome) and 6 healthy controls(HC) were entried to be evaluated the mechanism of various effects of acupuncture using thermographic method and blood examination of plasma ET-1, serotonin, nitrite and nitrate levels. Plasma ET-1 levels of PSS, SSSD and HC were 1.98𡟹0.69, 1.76𡟹0.39 and 1.15𡟹0.38 pg/ml, respectively. After the electrical acupuncture stimulation for unilateral side of hand/arm(30min.), the plasma ET-1 levels decreased in all 10 cases of PSS treated(1.61𡟹0.45 pg/ml), but no change of plasma serotonin levels in spite of increase of plasma nitrite levels in 2 cases of PSS. In 4 of 6 cases of SSSD and all 6 HC, plasma ET-1 levels increased(2.06𡟹0.39 pg/ml, 1.72𡟹0.58 pg/ml, respectively). The decrease of plasma ET-1 levels for the stimulation was thought to induce the vasodilatation and elevate the skin surface temperature in the patients with PSS.

-46- Matrix(2) 9/4 Room-C
TIMP-1 MAY BE AN AUTOCRINE GROWTH FACTOR IN SCLERODERMA FIBROBLASTS.
Kanako Kikuchi1, Takafumi Kadono2, Masutaka Furue1, KunihikoTamaki2.
1Department of Dermatology, Tokyo University Branch Hospital, 2Department of Dermatology, Faculty of Medicine, University of Tokyo, Tokyo, Japan.
We recently reported that the serum tissue inhibitors of metalloproteinase-1 (TIMP-1) levels in systemic sclorosis (SSc) patients were elevated compared with normal controls. To determine the biological significance of TIMP-1 in SSc, we compared the proliferative effects of TIMP-1 between normal and SSc fibroblasts. TIMP-1 showed significant mitogenic activity for both normal and SSc fibroblasts. The mitogenic responses to TIMP-1 (33-100 ng/ml) in SSc fibroblasts were significantly greater than those in normal controls, and were completely neutralized in the presence of anti-TIMP-1 IgG. Anti-TIMP-1 IgG partially but significantly blocked the basal mitogenic activities of SSc fibroblasts. SSc fibroblasts produced increased amounts of TIMP-1 relative to normal fibroblasts, as confirmed by Western blotting, ELISA and RT-PCR. Transforming growth factor- b1 (TGF-b1) up-regulated TIMP-1 production in normal fibroblasts but not in SSc fibroblasts with elevated spontaneous secretion of TIMP-1.

-47- Matrix(2) 9/4 Room-C
CONNECTIVE TISSUE GROWTH FACTOR GENE EXPRESSION IN TISSUE SECTIONS FROM EOSINOPHILIC FASCIITIS.
Takashi Kakinuma1, Nobukazu Hayashi2, Kanako Kikuchi1, Kunihiko Tamaki1, Atsuyuki Igarashi2.
1Department of Dermatology, Faculty of Medicine, University of Tokyo,2Division of Dermatology,Kanto Teishin Hospital,Tokyo,Japan.
Connective tissue growth factor(CTGF) was expressed in tissue regeneration and fibrotic diseases.We investigated CTGF gene expression in tissue sections from 6 patients with eosinophilic fasciitis using non-radioactive in situ hybridization. Digoxigenin-labeled CTGF mRNA probes were used.CTGF mRNA signals were observed in fascia of 2 patients who had remarkable inflammatory cell infiltration,and in deep dermis of 3 patients who had mild inflammatory cell infiltration,but the rest that demonstrated no inflammation had no signals.And 4 of 5 patients which expressed CTGF mRNA revealed high value of serum aldolase.In 2 cases of remarkable inflammatory cell infiltration,CTGF mRNA was expressed in fascia,and the value of serum aldolase was relative to CTGF gene expression.