사상체질의학회지 2010;22(4):56-64 J of Sasang Constitutional Medicine Vol. 22, No. 4, 2010:56-64 http://www.esasang.com 사상체질처방의항염증효능비교연구 이진아 하혜경 이호영 정다영 이준경 황대선 신현규 Abstract 한국한의학연구원한약 EBM 연구센터 Anti-inflammatory Activities of Herbal Formulas for Sasang Constitutional Medicine Jin-Ah Lee, Hyekyung Ha, Ho-Young Lee, Da Young Jung, Jun Kyoung Lee, Dae Sun Huang, Hyeun-Kyoo Shin Herbal Medicine EBM Research Center, Korea Institute of Oriental Medicine, Daejeon, 305-811, Korea 1. Objectives 4 herbal formulas (Yanggyeoksanhwa-tang, Yeoldahanso-tang, Cheongsimyeonja-tang and Taeeumjowi-tang) were applied to investigate the anti-inflammatory activities. In many studies, plant-derived anti-inflammatory efficacies have been investigate for their potential inhibitory effects on lipopolysaccharide (LPS)-stimulated macrophages. This study was performed to examine the anti-inflammatory activities of 4 herbal formulas on LPS-stimulated RAW 264.7 cells. 2. Methods The productions of nitric oxide (NO), prostaglandin (PG)E 2, interleukin(il)-6 and tumor necrosis factor (TNF)-α were examined in the presence of the 4 herbal formulas in RAW 264.7 cells. The cells were incubated with LPS 1 μg/ml and 4 herbal formulas for 18 hrs. The anti-inflammatory activity of 4 herbal formulas were investigate by carrageenin-induced paw edema in rats. The paw volume was measured at 2 and 4 hrs following carrageenininduced paw edema in rats. 3. Results Yanggyeoksanhwa-tang and Cheongsimyeonja-tang showed inhibitory effect on PGE 2 production in LPS-stimulated RAW 264.7 cells and a reduction in carrageenin-induced paw edema on rats. Yanggyeoksanhwa-tang showed inhibitory effect on IL-6 in LPS-stimulated RAW 264.7 cells. 4 herbal formulas not affect on NO and TNF-α inhibition in LPS-stimulated RAW 264.7 cells. 4. Conclusions These results suggested that Yanggyeoksanhwa-tang and Cheongsimyeonja-tang have anti-inflammatory activity. Key Words : Yanggyeoksanhwa-tang (Lianggesanhuo-tang), Yeoldahanso-tang (Reduohanshao-tang), Cheongsimyeonja-tang (Qingxinlianzi-tang), Taeeumjowi-tang (Taiyintiaowei-tang), Anti-inflammation 1) 접수일 2010 년 06 월 07 일 ; 심사일 2010 년 06 월 09 일 ; 승인일 2010 년 08 월 06 일 교신저자 : 신현규대전시유성구엑스포로 483 한국한의학연구원한약 EBM 연구센터 Tel : +82-42-868-9464 Fax : +82-42-864-2120 E-mail : hkshin@kiom.re.kr * 본연구는한국한의학연구원에서지원하는 표준한방처방 EBM 구축사업 에의해수행되었다.
이진아외사상체질의학회지 2010;22(4):56-64 57 Ⅰ. 緖論 사상체질의학에서四象處方은본초의溫熱凉寒의구성을사상체질이란시각으로대분류하고이들약물을體質證과病證에따라재구성하여이루어졌으며, 투약시병증에맞는처방을찾는것이최종목표가된다 1. 모든病證에는초기에염증반응이일어나며, 염증은생체조직의외부자극에대한방어반응의하나로서여러가지염증인자들이만들어지는데이로인하여임상적으로는발적, 발열, 종창, 동통, 기능장애등의증상이나타난다. 본연구에서는소양인처방인凉膈散火湯과태음인처방熱多寒少湯, 淸心蓮子湯, 太陰調胃湯등 4종의처방을대표적인사상체질처방으로선정하였다. 기존연구에서소양인처방인凉膈散火湯은뇌경색 2, 중풍환자 3 의증상을호전시키며, 혈당을조절하고 4 항아토피작용 5 이있다고연구되어있다. 태음인처방인熱多寒少湯은중풍환자의치료 6 및간기능에미치는영향 7 등이보고되었으며, 淸心蓮子湯은항노화효과 8 및항알레르기효과 9, 太陰調胃湯은소화기계기능강화 10 와항비만및항고지혈증에효과 11 가있다고알려져있다. Table 1. Combinations of Crude Drugs in Herbal Formulas Crude drug Componented crude drugs (g) Yanggyeoksanhwa-tang Yeoldahanso-tang Cheongsimyeonja-tang Taeeumjowi-tang 생지황 Rehmanniae Radix 7.5 인동등 Lonicerae Flos 7.5 연교 Forsythiae Frucus 7.5 산치자 Gardeniae Fructus 3.75 박하 Menthae Herba 3.75 지모 Anemarrhenae Rhizoma 3.75 석고 Gypsum 3.75 방풍 Saposhnikoviae Radix 3.75 형개 Schizonepetae Spica 3.75 갈근 Puerariae Radix 15.0 황금 Scutellariae Radix 7.5 3.75 고본 Angelicae Tenuissimae Radix 7.5 나복자 Raphani Semen 3.75 3.75 7.5 길경 Platycodonis Radix 3.75 3.75 승마 Cimicifugae Rhizoma 3.75 백지 Angelicae Dahuricae Radix 3.75 연자육 Nelumbinis Semen 7.5 산약 Dioscoreae Radix 7.5 천문동 Asparagi Radix 3.75 맥문동 Liriopis Tuber 3.75 3.75 원지 Polygalae Radix 3.75 석창포 Acori Graminei Rhizoma 3.75 3.75 산조인 Zizyphi Spinosi Semen 3.75 용안육 Longanae Arillus 3.75 백자인 Thujae Semen 3.75 감국화 Chrysanthemi Flos 3.75 의이인 Cocicis Semen 11.25 건율 Castaneae Semen 11.25 오미자 Schizandrae Fructus 3.75 마황 Ephedrae Herba 3.75 Total (g) 45 45 52.5 48.75 Yield (%) 16.2 29.0 20.5 19.9
58 사상체질의학회지 2010;22(4):56-64 사상체질처방의항염증효능비교연구 모든질병은초기에기본적으로염증반응이관여되는데, 이들처방이인체의항염효과에대한연구는없어, 본연구에서는사상체질처방중대표적으로사용빈도가높은 4종처방을선정하여항염증활성을비교하였다. Ⅱ. 材料및方法 1. 재료 1) 처방추출본실험에사용된 4종의처방구성은 Table 1과같으며처방구성약재들은 ( 주 ) 옴니허브 (Yeongcheon, Korea) 와 ( 주 )HMAX (Chungbuk, Korea) 에서각각구입하였다. 각각의처방에따라약재를배합한후이의 10배의물과함께 100 에서 2시간동안무압환류추출법 ( 추출기 : 경서메디텍 COSMOS660) 을이용하여추출하였다. 추출액은 sieve를사용하여거른후동결건조기 ( 일신동결건조기 PVTFD100R) 를사용하여처방추출물분말을얻었으며수득율은 Table 1과같다. 2. 실험방법 1) RAW 264.7 세포주배양생쥐대식세포주인 RAW 264.7 세포주는 American Type Culture Collection (ATCC, USA) 에서분양받아사용하였다. RAW 264.7 세포는 5.5% fetal bovine serum (FBS; Gibco, USA), 1% penicillin-streptomycin (Gibco, USA) 이첨가된 Dulbecco's modified eagle's medium (DMEM; Gibco, USA) 배지에 37, 5% CO 2 조건에서배양하였다. 2) 세포독성측정 4종처방추출물의세포독성을알아보기위해 cell counting kit-8 (CCK-8; Dojindo, Japan) 을제조사의사용법에따라사용하였다. 96 well plate에 5 10 3 cells/well 씩분주한세포에 4종처방추출물을농도별로처리하여 24시간동안배양하였다. CCK-8 용액 10 μl를첨가하여 4시간동안배양한후 450 nm에서흡광도를측정하고대조군과의비교를통해상대적인세포생존율 (% of control) 을계산하였다. 이후의실험은세포독성 이나타나지않는최고농도를기준으로실험하였다. 3) NO, PGE 2, IL-6, TNF-α 생성량측정 2.5 10 5 cells/well (48 well plate) 로분주한 RAW 264.7 세포를 LPS (1 μg/ml) 로자극하고, 4종처방추출물을농도별로처리하여 18시간동안배양하였다. 생성된 NO의양을측정하기위해배양상층액의 nitrite 양을 Griess reagent (Promega, USA) 를사용하여제조사의사용법에따라측정하였으며양성대조군으로 NOS inhibitor인 NG-methyl-L-arginine (L-NMMA; Sigma, USA) 를사용하였다. PGE 2 양을측정하기위해 Arachidonic acid (Sigma, USA) 를최종 30 μm이되도록처리해 15분간반응시킨후배양상층액을취하여 enzyme-linked immunosorbent assay (ELISA) kit (Amersharm, UK) 를사용하여제조사의사용법에따라측정하였으며양성대조군으로 indomethacin을사용하였다. IL-6와 TNF-α 생성량을측정하기위해배양상층액을취하여 ELISA kit (R&D, USA) 를사용하여제조사의사용법에따라 IL-6와 TNF-α양을측정하였다. LPS군과의비교를통해상대적인비율 (% of LPS) 을계산하였다. 4) 실험동물생후 4주된수컷 SD 랫트 ( 오리엔트바이오, 성남 ) 를분양받아온도 23±3, 상대습도 50±10%, 12시간간격으로명암이조절되는동물사육실에서 1주일간환경에적응하도록순화시켰다. 순화기간중육안적으로관찰하여건강한동물을선별하여실험에사용하였다. 5) Carrageenin 유발족부종모델수컷 SD 랫트에게 7일간 4종처방추출물 1 g/kg/day 를경구투여하고, 랫트의오른쪽발바닥피하에 1% carrageenin (Sigma, USA) 생리식염액을 0.1 ml씩주사하여족부종을유발하였다. 발생된족부종을수용적법으로 carrageenin 주사직전과주사 2시간및 4시간후 plethysmometer (Ugo Basile, Italy) 를이용하여측정하였다. 측정한발의부피를이용하여산출법에의하여부종증가율을계산하였으며양성대조군으로 indomethacin (10 mg/kg/day) 을단회투여하여사용하였
이진아외사상체질의학회지 2010;22(4):56-64 59 Figure 1. Effects of herbal formulas on LPS-stimulated NO production in RAW 264.7 cells. Cells were treated with herbal formulas and cultured for 18 hr with LPS (1 μg/ml). The NO production in cell culture supernatants were measured as described in Materials and Methods. The units of concentrations of L-NMMA and herbal formulas are μm and μg/ml, respectively. The data are presented as mean±s.e.m. and are representative of triplicate experiments; # p<.050 compared to LPS-free cultures (NC; untreated control cell), * p<.050 compared to cells cultured with LPS. 다. 산출법은다음과같다. 부종증가율 (%) = 족부종유발후발의부피 족부종유발전발의부피 100 족부종유발전발의부피 6) 통계처리모든측정결과는 mean±s.e.m. 으로나타냈으며실험군간의차이는 ANOVA를사용하여통계학적분석을수행하였으며 p<0.050일경우에유의성이있다고하였다. 미치는영향을확인하고자 RAW 264.7 세포에 LPS를처리하여염증반응을유발시키고, 4종처방추출물을처리하여 NO 생성량을측정하였다. 양성대조군으로사용한 NOS inhibitor인 L-NMMA는농도의존적으로 NO 생성을억제하는것으로나타났다. NO 생성은凉膈散火湯이 100 μg/ml에서 29.2% (p<.050), 熱多寒少湯이 100 μg/ml에서 35.9% (p<.050), 淸心蓮子湯이 200 μg/ml에서 27.6% 억제하였다 (p<.050). 그러나太陰調胃湯은 NO 생성억제효과를나타내지않았다 (Figure. 1). Ⅲ. 結果 1. 대식세포독성 RAW 264.7 세포에 4종처방추출물을처리하고, 24시간동안배양한후, 세포독성을관찰한결과, 凉膈散火湯, 熱多寒少湯은 100 μg/ml까지, 淸心蓮子湯, 太陰調胃湯 200 μg/ml까지세포독성이나타나지않았다. 따라서모든실험을세포생존에영향을주지않는범위내에서수행하였다 (do not shown). 2. NO 생성억제효과 4종처방이대식세포에서 LPS에의한 NO 생성에 3. PGE 2 생성억제효과 4종처방의 PGE 2 생성억제효과를검색하고자 RAW 264.7 세포에 LPS를처리하여염증반응을유발시키고, 4종처방추출물을처리하여 PGE 2 생성량을측정하였다. 그결과, 양성대조군으로사용한 indomethacin 은농도의존적으로 PGE 2 생성을억제하는것으로나타났다. 凉膈散火湯은 PGE 2 생성을농도의존적으로억제하여 50 μg/ml에서 LPS에비해 27.6% 억제하였으나유의하진않았다. 淸心蓮子湯은 PGE 2 생성을농도의존적으로억제하여 200 μg/ml에서 LPS에비해 32% 억제하였다 (p<.050). 그러나熱多寒少湯, 太陰調胃湯은 PGE 2 생성억제효과가없었다 (Figure. 2).
60 사상체질의학회지 2010;22(4):56-64 사상체질처방의항염증효능비교연구 Figure 2. Effects of herbal formulas on LPS-stimulated PGE 2 production in RAW 264.7 cells. Cells were treated with herbal formulas and cultured for 18 hr with LPS (1 μg/ml). The PGE 2 production in cell culture supernatants were measured as described in Materials and Methods. The units of concentrations of indomethacin and herbal formulas are ng/ml and μg/ml, respectively. The data are presented as mean±s.e.m. and are representative of triplicate experiments; # p<.050 compared to LPS-free cultures (NC; untreated control cell), * p<.050 compared to cells cultured with LPS. Figure 3. Effects of herbal formulas on LPS-stimulated IL-6 production in RAW 264.7 cells. Cells were treated with herbal formulas and cultured for 18 hr with LPS (1 μg/ml). The IL-6 production in cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA) kit, as described in Materials and Methods. The units of concentrations of indomethacin and herbal formulas are ng/ml and μg/ml, respectively. The data are presented as mean±s.e.m. and are representative of triplicate experiments; # p<.050 compared to LPS-free cultures (NC; untreated control cell), * p<.050 compared to cells cultured with LPS. 4. IL-6및 TNF-α 생성억제효과 NO와 PGE 2 생성억제를나타낸凉膈散火湯, 淸心蓮子湯에대한 IL-6와 TNF-α 생성억제효과를검색하고자 RAW 264.7 세포에 LPS를처리하여염증반응을유발시키고, 2종처방추출물을처리하여 IL-6 와 TNF-α 생성량을측정하였다. 그결과凉膈散火湯은 IL-6 생성을농도의존적으로억제하여 50 μg/ml에서 29.9% 억제하였고 (p<.050), 淸心蓮子湯은 IL-6 생성에대한억제효과가없었다 (Figure. 3). 凉膈散火湯, 淸心蓮子湯모두 TNF-α 억제효과가없었다.
이진아외사상체질의학회지 2010;22(4):56-64 61 Figure 4. Effects of herbal formulas on carrageenin-stimulated paw edema in rats. The control group (0.1% Tween 80) and herbal formulas (1 g/kg/day) were orally administered for seven days prior to the inception of carrageenin-stimulated paw edema. The imdomethacin group (10 mg/kg/day) was orally administered for 3 hr prior to the inception of carrageenin-stimulated paw edema. The paw volume was measured as described in Materials and Methods. The data are presented as mean±s.e.m. (5 rats per group). 5. Carrageenin 유발족부종모델 NO, PGE 2, IL-6 생성억제를나타낸凉膈散火湯, 淸心蓮子湯에대한 carrageenin 유발족부종모델에서의항부종효과를검색하기위하여 2종처방추출물을투여하고 carrageenin으로족부종을유발한후발의부피를측정한결과정상상태보다족부종유발후발의부피가 2시간, 4시간째각각 28.5, 31.8% 증가되었다. 양성대조군으로사용한 indomethacin 투여군은족부종유발군보다 2시간및 4시간에각각 45.8%, 53.6% 감소되었다. 凉膈散火湯투여군은족부종유발군보다 2시간후 34% 억제하였으나 4시간후는 19% 억제하였으나이는유의하진않았다. 淸心蓮子湯투여군은족부종유발군보다 2시간및 4시간에각각 35%, 40.3% 로부종이감소되었으나유의하진않았다 (Figure. 4). Ⅳ. 考察 질병은초기부터염증반응이시작되어증상을발현하기시작한다. 사상체질의학에서사용되는처방들도체질에따라처방이분류되기는하지만이들염증에대한항염증작용이필요하다고판단되어사상체질처방중에대표적인처방을 4종선정하여항염증효과를확인하고자하였다. 대표적인사상체질처방으로선정한凉膈散火湯 은소양인처방으로 東醫壽世保元 에처음으로수록되었으며소양인胃受熱褢熱病의胸膈熱證에대한淸陽上升을목적으로사용되었고 12, 임상적으로는胸悶, 手足痹, 運動障碍, 手足麻木, 火熱證, 口舌疾患, 上焦熱皮膚疾患등에다용되고있다 13. 태음인처방중熱多寒少湯은肝受熱褢熱病에사용되는처방으로主治는肝熱熱症溫燥熱病手指炒黑癍瘡病虛勞夢泄이다. 淸心蓮子湯은肝受熱褢熱病에사용되는처방으로 東醫壽世保元 에서는처방구성내용만제시했을뿐主治症이나治驗例및活用法에대한언급은없으며 東醫四象新編 에서虛勞, 夢泄, 無度, 腹痛, 泄瀉, 舌卷, 中風, 食滯, 胸膈痛등을치료한다고기록되어있다 14. 太陰調胃湯은 黃疸傷寒時氣頭痛身痛無汗食滯痞滿腿脚無力 등의太陰人表寒病에응용되는처방으로기술되어있다. 12 이들처방의항염효능을검증하기위한기존연구는아직미비한실정이다. 염증은생체조직의외부자극에대한방어반응의하나로서염증반응이일어나면여러가지염증인자들이만들어지는데이로인하여임상적으로는발적, 발열, 종창, 동통, 기능장애등의증상이나타난다. 염증인자에는 inducible nitric oxide synthase (inos) 에의해서만들어지는 nitric oxide (NO) 와 cyclooxygenase-2 (COX-2) 에의해서만들어지는 prostaglandin E 2 (PGE 2) 등이있다 15. 이러한염증인자는염증반응의전사인자인 nu-
62 사상체질의학회지 2010;22(4):56-64 사상체질처방의항염증효능비교연구 clear factor-κb (NF-κB) 를활성화시켜 NO와 PGE 2 를생성하여염증을일으킨다 16, 17. NOS에의해 L-argine 으로부터생성되는 NO는 18 혈관투과성, 부종등의염증반응을촉진시킬뿐만아니라염증매개체의생합성을촉진하여염증을심화시킨다 19. 특히대식세포가 LPS로자극될때 inos가발현되어 NO를생성하게된다 20. 본연구에서凉膈散火湯은 100 μg/ml, 熱多寒少湯은 100 μg/ml, 淸心蓮子湯 200 μg/ml에서 LPS 처리군보다각각 29.2%, 35.9%, 27.6% NO 생성을억제하였으나太陰調胃湯는 NO 생성억제효과가없었다. COX는 arachidonic acid를 PG로전환하는효소로혈소판형성, 위벽보호및신장기능의유지등정상적인생리기능에작용하는 COX-1과염증매개물질인 PGE 2 를생성하는 COX-2로분류된다 21. 다수의항염증제의작용기전은 PG 합성을억제하는것인데이것은 COX-2의활성저해에의한것이다. PGE 2 는면역관련세포의활성을유도하여염증반응을항진시켜염증성사이토카인을대량생성하게한다 22. 4종처방의 PGE 2 생성억제를검색한결과淸心蓮子湯은 200 μg/ml에서 LPS 처리군보다 PGE 2 생성을 32% 억제하였고 (p<0.05) 凉膈散火湯은 50 μg/ml에서 LPS 처리군보다 27.6% 억제하였으나유의하진않았다. 그리고熱多寒少湯, 太陰調胃湯은효과를나타내지않았다. 대식세포는초기염증반응에관여하는대표적인면역세포로서염증자극시염증성사이토카인인 IL-6, TNF-α 등을생성한다 23. 이러한염증성사이토카인들은면역세포를활성화시켜여러가지염증성질환에관여한다. NO와 PGE 2 생성억제효과를나타낸凉膈散火湯은농도의존적으로 IL-6 생성을억제하였으나淸心蓮子湯는억제효과가없었으며凉膈散火湯, 淸心蓮子湯모두 TNF-α의생성에대한억제효과가없었다. 생체내에서의항염증활성을검색하기위하여 carrageenin 유발족부종모델을사용하였다. Carrageenin 은홍조류로부터얻어진 polysaccharide로서급성염증유발제로항염증제의탐색을위한실험모델에빈번하게사용되고있다 24, 25. Carrageenin에의해발생하는부종을억제하는항염증제의작용기전은 COX의활 성저해와관련성이있을것으로보고되고있다 26. 대표적인항염증제인 indomethacin은 PGs의합성을억제하는것으로알려져있으며 27, 본연구에서도 carrageenin에의한족부종을감약시켰다. 凉膈散火湯, 淸心蓮子湯 2종의처방은족부종을감소시켰으나유의하진않았다. 이는 COX의활성저해로인한 PGE 2 생성억제로인해감소되었을것으로생각된다. 이상의결과를종합해보았을때, 사상체질처방중凉膈散火湯은대식세포의염증반응에서 NO, PGE 2, IL-6 생성량및족부종을억제시켰고, 淸心蓮子湯은대식세포의염증반응에서 NO, PGE 2 생성량및족부종을억제시킴으로서凉膈散火湯, 淸心蓮子湯이 inos와 COX에의한염증매개물질생성을억제하는항염증활성이있음을확인할수있었다. 따라서본연구결과는소양인처방인凉膈散火湯과태음인처방인淸心蓮子湯이앞으로염증성질환등의치료에유용하게사용될수있다는기초자료로활용될수있을것으로기대된다. Ⅴ. 結論 소양인처방인凉膈散火湯과태음인처방인熱多寒少湯, 淸心蓮子湯, 太陰調胃湯등 4종처방의항염증활성을연구한결론은다음과같다. 1. 대식세포에서 LPS로염증반응을유발하여 NO를측정한결과, 凉膈散火湯, 熱多寒少湯, 淸心蓮子湯 3종의처방은 NO 생성을억제하였으나太陰調胃湯는 NO 생성억제효과가없었다. 2. 대식세포에 LPS로염증반응을유발하여 PGE 2 를측정한결과, 凉膈散火湯, 淸心蓮子湯은 PGE 2 생성을억제하였으나熱多寒少湯, 太陰調胃湯은억제효과가없었다. 3. 대식세포에 LPS로염증반응을유발하여염증성사이토카인인 IL-6와 TNF-α를측정한결과, 凉膈散火湯은 IL-6 생성을억제하였으나淸心蓮子湯은효과가없었다. 凉膈散火湯, 淸心蓮子湯모두 TNF-α 생성억제효과가없었다.
이진아외사상체질의학회지 2010;22(4):56-64 63 4. 凉膈散火湯은족부종유발 2시간후족부종유발군에비해족부종이억제되었고, 淸心蓮子湯은족부종유발 2시간및 4시간후족부종유발군에비해족부종이억제되었다. 이상의결과로凉膈散火湯, 淸心蓮子湯은항염증활성이있는것으로판단된다. Ⅵ. 感謝의글 본연구는한국한의학연구원에서지원하는 표준한방처방 EBM 구축사업 (K10030) 에의해수행되었으며, 이에감사드립니다. Ⅶ. 參考文獻 1. Dept. of Sasang constitutional medicine. Sasang Constitutional Medicine. Seoul: Gibmundang. 2006. 347, 357, 420 (Korean) 2. Lo HS, Lee SM, Bae YC, Park HS, Lee JS, Song SG, et al. The effect of Yangkyuk-Sanhwa-Tang on cytokine production in the patients with cerebral infarction. J Sasang Constitut Med. 2004;16(1):120-129. (Korean) 3. Choi DJ, Ryu SH, Jung WS, Moon SK, Cho KH, Kim YS. The Clinical Efficacy of Yangkyuksanwha-tang on acute stroke. J Korean Oriental Med. 2004;25(1): 111-116. (Korean) 4. Jung SI, Kim JW. Clinical Study about the Diabetes Mellitus Patients Administration of Yangkyuksanwhatang. Korea J Oriental Physiology & Pathology. 2002; 16(6):1308-1313. (Korean) 5. Yun BH, Park SS. Yangkyuksanhwa-tang effected to atopic dermatitis. J Sasang Constitut Med. 2004; 16(2):84-98. (Korean) 6. Choi JS, Kim KY, Kim HM, Ju JC. Studies on the cytokine production regulation in human astrocytes by Yuldahansotang. J Sasang Constitut Med. 2001; 13(1):61-69. (Korean) 7. Kim TH, Hong SY, Han DN, Lim EC. The Effects of constant use of Yeoldahanso-tang on liver function. J Sasang Constitut Med. 2007;19(3):217-226. (Korean) 8. Lim JP, Ahn TW. The anti-oxidative and immune-regluatory effect of Chungsimyeonja-tang in aged rat. J Sasang Constitut Med. 2007;19(3):227-241.(Korean) 9. Park SC. Effects CheongSimYeonJaTang(CSYJT) on control of immune-function in highly purified mouse B cells and Mast cell. J Sasang Constitut Med. 2003;15 (2):166-179. (Korean) 10. Chou LS, Ahn TW, Lee H. Studies of Chengsimyeonjatang and Taeumjowi-tang that get change by Wistar rat's aging. J Sasang Constitut Med. 2005;17(2):74-84. (Korean) 11. Kim KS, Lee DU, Kim YL, Hwang MJ, Kim GW, Koo BS. Anti-obesity and anti-hyperlipidemic effects of Taeyeumjowee-tang and its modified prescription. J Oriental Neuropsychiatry. 2007;18(2):57-74. (Korean) 12. Lee JM. Dongeuisusebowon. Seoul:Yeogang Publishing Co. 1992:271-272.(Korean) 13. Yun BH, Park SS. Yangkyuksanhwa-tang effected to atopic dermatitis. J Sasang Constitut Med. 2004; 16(2):84-98. (Korean) 14. Won JS. Dongeuisasangshinpyeon. Seoul:Munw- oosa. 1926:2. 15. Posadas I, Terencio MC, Guillén I, Ferrándiz ML, Coloma J, Payá M et al. Co-regulation between cyclo-oxygenase-2 and inducible nitric oxide synthase expression in the time-course of murine inflammation. Naunyn Schmiedebergs Arch Pharmacol. 2000;361(1): 98-106. 16. Lee TH, Kwak HB, Kim HH, Lee ZH, Chung DK, Baek NI et al. Methanol extracts of Stewartia koreana inhibit cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (inos) gene expression by blocking NF-kappa B transactivation in LPS-activated RAW 264.7 cells. Mol Cells. 2007;23(3):398-404. 17. Nishida T, Yabe Y, Fu HY, Hayashi Y, Asahi K, Eguchi H et al. Geranylgeranylacetone induces cyclooxygenase-2 expression in cultured rat gastric epithelial cells through NF-kappaB. Dig Dis Sci. 2007;52(8): 1890-1896.
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