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1 Korean Chem. Eng. Res., Vol. 44, No. 1, February, 2006, pp 치마버섯유래다당체의화상및상처치유효과 Ç h o o }e q ( o 26p r, o 31p }ˆ) Effect of Polysaccharide from Schizophyllum commune on Burn and Wound Healing Min Kyoung Kim and Eock Kee Hong Department of Bioengineering & Technology, Kangwon National University, 192-1, Hyoja-2-dong, Chuncheon, Kangwondo , Korea (Received 26 December 2005; accepted 31 January 2006) k o ~ k l p n n v p. p ~ p }l p k sv r m sv q l l l~ eˆp f o ˆo k v p. np ~ q d n l q svp 2 m p lr p. l l d m l p n o ~ pn l in vivo e k r l } o Š m. l 2 mp o o p o r Š l p p m. h Abstracts Polysaccharides extracted from mushroom have been most commonly used to enhance the immune system. Also polysaccharides maintaining the moisture extent on epidermal tissue have an effect on the removal of necrotic tissue and the restoration of epidermal tissue through enhancing the immune system at skin layers. In this work, polysaccharides were from Schizophyllum commune studied about the burn and wound healing activity in the epidermal tissue on rats through in vivo experiment and hematological values. And antibacterial activities were examined using pathogenic microorganisms causing the secondary inflammation. Key words: Polysaccharide, Burn and Wound Healing, Schizophyllum commune 1. }l qp psp on p l k qnl ~ rp l v l k, t d r, k, p d, m l p l p [1, 2]. o p β-glucanp } ol rp qpqp p v t q pq p p v l q p v eˆ p pv l [3]., o p β-glucanp p l m k q s e p q p eˆ svp pme v p eˆ ( v)p p lr pp 2 mp pp } o p eˆ p p., p o p s, p ˆ p o To whom correspondence should be addressed. ekhong@kangwon.ac.kr ve t l v p k r p [4]. q n p v l l r, llrr e p pp p l p r p v l n. m l, } k p v l n p l rm llrr p ~ l p p p. l rp qnp l r l p e, r, v p ppˆ p p o v p m p svl e p r l 2 mp mv pp lr ~ p ql o l p s l l v p eˆ q p. v l r rp v p lr l ql o p p r r p p p k. } p v l pl e m el eˆ p r n p [5-7]. l l p llv ~ pn l } o in vivo e k r l 87

2 88 Ë l Š mp, Staphylococcus sp. p 2 mp o p p l kk q m m e l n }l p k~ k lkp lˆm r Œ p l rs m. (t) k lp Fucidin (t)ˆ k lp Azulene-Sm p nl k s l rn sl l e l rs l m l } ol p Š m., ~p p rl nl ((t) rk) l n m. } r l e e u l k p H1-system q (Technicon Co.) n m., rl p bacteriap Bacillus megaterium(kctc 3709), Staphylococcus aureus(kctc 1621), Streptococcus pyogenes(kctc 3096)m yeastp Candida albicans (KCTC 7270) pn l e p m i oo k~ k lkp llv ~ r n o ~p 100 mg/mlp ˆpm l p, l l nl 100 µlj l l skrm ~ l, ~ l rs m m lo e p (t)m l 6t (200~250 g)p o Sprague-Dawley k k m 22±3 o C, d 50±5Í, 10~15 /p, 12 h t k, s 150~300 Luxp o l ptp rp e p, (t)m l n p pn m. }p p 7p m. e p e Table 1l ˆ l zm z 6t ratp ol p r, p p pr, 30 l p k l. l p s p k } v kp, s krp Azulene-S, skrm ~ }, ~ } s p p } p l m. l p e (Azulene-S n) } p e (Fucidin n) p p m. p l } pp o p r m n y} h n e rat urethanep eˆ l l k 10 mlp } m. r p } EDTA p r lp tube pn l } m. } r p q pn l k p (white blood cells, WBC), r (red blood cells, RBC), (hemoglobin, Hb), r nr (hematocrit, HCT) Ž (platelet, PLT) r m ~o Bacteriap o v Nutrient Broth(beef extract 3 g/l, peptone 5g/l) n m, yeastp Candida albicansp n glucose(40 g/l), peptone(10 g/l), yeast extract(2 g/l) v pn m. k m Bacillus megateriump 30 o C, bacteriap n 37 o C pl, Candida albicansp n 24 o C pl. r l n s v ~ l v kp v n l p agar plate t m. e v ~p 10 mg/ml, 1 mg/ml, 0.1 mg/ml, 0.01 mg/mlp vl ~ l rs m. r kp ~ l pv kp plate l k t p v 50 mlp kkl single colony rs 12e k k m. kk 100 µl l plate 200~300 p p r p pt 100 µl plate l rs l spreading m. 24e k platel p colony Š l p r m. 3. y 3-1. os {s l p p pp e ratp n, n l r(azulene-s) p } k s o p ~m } e l o skp p okp p m (Fig. 1, Fig. 2). e l v ol mv p mp ve(hyperplasia of epidermis)p pl. s l p plp e l pv kk. v svl o mvp k plp v (sebaceous gland)p l p Ž l. v l p p n s l ee r e l ƒ r opr p l o. e l p r o ˆo p Ž l., } p e l r rp p q l š v s l r(fucidin)m ~ } nl k } v kp e sl r p } e }p o n p pl (Fig. 3). p o p ~ } o Table 1. Grouping of rats for artificial experiments Experimental group scalded o44 o k Normal control No treatment Azulene-S or Fucidin Azulene-S or Fucidin + polysaccharide Vaseline + polysaccharide Experimental group burned Experimental group wounded Vaseline

~p } o 89 Fig. 1. Experimental groups scalded in the back. A. initial control, B. control after 7 days, C. rat treated with Azulene-S, D. rat treated with Azulene-S+polysaccharide, E.

rat treated with Fucidin, D. rat treated with Fucidin+polysaccharide, E. rat treated with vaseline, F. rat treated with vaseline+polysaccharide(after 7 days).

rat treated with Azulene-S, D. rat treated with Azulene-S+polysaccharide, E. rat treated with vaseline, F. rat treated with vaseline+polysaccharide(after 7 days). 3-2.

3 ~p } o 89 Fig. 1. Experimental groups scalded in the back. A. initial control, B. control after 7 days, C. rat treated with Azulene-S, D. rat treated with Azulene-S+polysaccharide, E. rat treated with vaseline, F. rat treated with Vaseline+polysaccharide(after 7 days). Fig. 3. Experimental groups wounded in the back. A. initial control, B. control after 7 days, C. rat treated with Fucidin, D. rat treated with Fucidin+polysaccharide, E. rat treated with vaseline, F. rat treated with vaseline+polysaccharide(after 7 days). l e n qpqp p ve ~ p p β-glucanp }svp l pp eˆ p Ž [8]., } ol do p s l 2 mp lr p f } ol rpl Ž. Fig. 2. Experimental groups burned in the back. A. initial control, B. control after 7 days, C. rat treated with Azulene-S, D. rat treated with Azulene-S+polysaccharide, E. rat treated with vaseline, F. rat treated with vaseline+polysaccharide(after 7 days) h n p p e l ~ pn l l r } up k j v dp m (Table 2). p l r p mv o p pv, p r op 6.60~ /µll l v kpp p p l., r m p n l rp Azulene-Sp } nl k p pl, p o svp p p Ž lp [9], Žp ~rp r o ov p p ˆ. p p e p n, r o l n pp r rp r o ov m. t } e l e pp l mv p pp m, e p r m Ž, r tpl. } p e p n etl n p l r(fucidin) } Korean Chem. Eng. Res., Vol. 44, No. 1, February, 2006

4 90 Ë l Table 2. Hematological value of the rats during burn and wound experimental periods Groups WBC ( 10 3 /µl) RBC ( 10 3 /µl) Hb (g/dl) HCT ( ) PLTG( 10 3 /µl) Normal range 6.6~ ~ ~ ~ ~450 Standard group Normal control 9.02± ± ± ± ±2.121 Experimental group Control 5.66± ± ± ± ± scalded Azulene-S 8.47± ± ± ± ±24.74 Vaseline 7.62± ± ± ± ±24.04 Azulene-S+polysaccharide 11.32± ± ± ± ± Vaseline+polysaccharide 8.08± ± ± ± ±36.06 Experimental group Control 9.07± ± ± ± ± burned Azulene-S 10.43± ± ± ± ±4.941 Vaseline 13.94± ± ± ± ±36.06 Azulene-S+polysaccharide 13.09± ± ± ± ± Vaseline+polysaccharide 12.75± ± ± ± ±11.31 Experimental group Control 7.18± ± ± ± ±4.941 wounded Fucidin 14.55± ± ± ± ±25.45 Vaseline 19.2± ± ± ± ±2.121 Fucidin+polysaccharide 7.85± ± ± ± ±67.88 Vaseline+polysaccharide 7.72± ± ± ± ±54.44 Table 3. Antibacterial activity of polysaccharide in agar plate ColonyW Control a b c d # Strains Colony # Colony # IR( ) Colony # IR( ) Colony # IR( ) Colony # IR( ) Bacillus megaterium Staphylococcus aureus Streptococcus pyogenes Candida albicans *Control: medium not including polysaccharide a. Polysaccharide concentration: 10 mg/ml, b. Polysaccharide concentration: 1 mg/ml, c. Polysaccharide concentration: 0.1 mg/ml, d. Polysaccharide concentration: 0.01 mg/ml *IR: Inhibition rate( ) e p r o r l mv o [10]p p l l ~ pn l l n nl, r m Ž, r o ov p p m ~ agar platel q p colony ~ v kp control platel p colony r ˆ p plp, ~p kp p o vp p colony r ˆ p pl (Table 3). Staphylococcus aureusm Streptococcus pyogenesp n v ~p 0.1 mg/ml, 0.01 mg/mlp platel p colony p p l ˆ v kv, ~ o n n n p qp p ˆ., Bacillus megateriumm Candida albicansp nl r~rp controll l qp r l p k pl [11]. 4. }l p } p r p p p o e rat pn l e r } p o44 o k in vivo e m k r p ee l } o Š m. n l p } p u o p ~ o l p l o, skr n k } v kp s l l } o lpp l pl. } p u p p v k p p plp, l o p p pl. p p e p } o s kr l ~p d l p p Ž. k r, r~rp o p ~ l } e p n kp r ol l v k pp k plp, }o e l mv p p rp p p pl. p f ~ l sp l r }l o m 2 mp m l ppp p m. mvp o p p ~p kk, ~ o l p vl q p s l qp r k. Bacillus megaterium Candida albicansp qp e r p e p l. rp o ~ } o k m ˆ p p pl.

5 y ~p } o DiLuzio, N. R., Immunopharmacology of Glucan: a Broad Spectrum Enhancer of Host Defense Mechanisms, Trends in Pharm. Sci., 4, (1983). 2. Chihara, G., Hamuro, J., Maeda, Y., Arai, Y. and Fukuoka, F., Fractionation and Purification of the Polysaccharides with Marked Antitumor Activity, Especially Lentinan, from Lentinus edodes, (Berk) Sing, (an Edible Mushroom), Cancer Res., 30(11), (1970). 3. Maeda, Y. and Chihara, G., Lentinan, a New Immuno-accelerator of Cell-mediated Response, Nature, 229(5287), (1971). 4. Tsukagoshi, S. and Ophashi, F., Protein-Bound Polysaccharide Preparation, PS-K, Effective Against Mouse Sarcoma-180 and rat Ascites Hepatoma AH-13 by oral use, Gann., 65(6), (1974). 5. Wasser, S. P. and Weis, A. L., Therapeutic Effect of Substances Occurring in Higher Basidomycetes Mushrooms: A Modern Perspective, Crit. Rev. Immunol., 19(1), 65-96(1999). 6. Ohno, N., Miura, N. N., Chiba, N., Adachi, Y. and Yadomae, T., Comparison of the Immunopharmacological Activities of Triple and Single-helical Schizophyllan in Mice, Biol. Pharm. Bull., 18(9), (1995). 7. Kim, M. S., Park, K. M., Chang, I. S., Kang, H. H. and Sim, Y. C., β-(1,6)-branched β-(1,3)-glucan in Skin Care, Allured s Cosmetics & Tolletries, 115(7), 79-86(2000). 8. Su, C. H., Sun, C. S., Juan, S. W., Ho, H. O., Hu, C. H. and Sheu, M. T., Development of Fungal Mycelia as Skin Substitutes: Effects on Wound Healing and Fibroblast, Biomaterials, 20(1), 61-68(1999). 9. Muzzarelli, R. A., Baldassare, A., Conti, V. F., Ferrara, P., Gazzanelli, G. and Vasi, V., Biological Activity of Chitosan: Ultrastructural Study, Biomaterials, 9(3), (1988). 10. Hwang, S. K., Kim, Y. Y., Lee, H. S., Bae, J. S. and Kim, D. Y., Burn Remedial Effects on Rat Skins of Natural Polysaccharide Ointments, J. Korean Ind. Eng. Chem., 11(6), (2000). 11. Hatvani, N., Antibacterial Effect of the Culture Fluid of Lentinus edodes Mycelium Grown in Submerged Liquid Culture, Intl. J. Antimicrobial Agents, 17(1), 71-74(2001). Korean Chem. Eng. Res., Vol. 44, No. 1, February, 2006

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<30372E31362D323028BDC5C7F6C5C32DB9CCB1B970626D292E666D> k m v r1 r1 16~20 (2009) J. Korean Academy of Managed Care Pharmacy Vol. 1, No. 1 Pharmacy Benefit Management iç 1# x sƒ, 1 w w w Overview on Pharmacy Benefit Management in USA Kyung Eob ChoiGand Hyun