Korean J. Plant Res. 28(4): (2015) Print ISSN Online ISSN Original Research Art

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1 Korean J. Plant Res. 28(4): (2015) Print ISSN Online ISSN Original Research Article 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 김민경 1, 이지선 2, 박영철 2, 최선미 1, 이상훈 1 * 1 한국한의학연구원한의기반연구부, 2 대구카톨릭대학교 GLP 센터 A 28 Day Repeated Dose-Oral Toxicity Studies of Arisaema Rhizome Aqueous Extracts in Sprague-Dawley Rats Min-Kyeoung Kim 1, Ji Sun Lee 2, Yeong Chul Park 2, Sun Mi Choi 1 and Sanghun Lee 1 * 1 KM Fundamental Research Division, Korea Institute of Oriental Medicine, Daejeon , Korea 2 GLP Center, Catholic University of Daegu, Keongsan , Korea Abstract - The object of this study was to obtain single oral dose toxicity of Arisaema Rhizome (Arisaema amurense f. serratum (Nakai) Kitag) aqueous extracts. Arisaema Rhizome (Chunnamsong in Korean) is one of the most important folk remedy plants used in Asia. In the study, a 28-day rat oral gavage study has been conducted with the extracts from Arisaema Rhizome at dose of 1,250, 2,500 and 5,000 mg / kg /day. The following endpoints were evaluated: clinical observations, body weight, gross and microscopic pathology, clinical chemistry, and hematology. Based on the analysis of these endpoints, it was estimated that NOEL (no observed effect level) for male rats and NOAEL (no observed adverse effect level) for female rats are 5000 mg / kg /day of the water-extracts from Arisaema Rhizome. Key words - Arisaema Rhizome, Toxicity test, NOEL, NOAEL, Folk remedy 서언 한약재천남성은천남성과 (Araceae) 에속한다년생식물인 둥근잎천남성 (Arisaemaamurense Maxim.), 천남성 (Arisaema amurense f. serratum (Nakai) Kitag) 또는두루미천남성 (Arisaema heterophllum Blume) 의주피를완전히제거한덩이 뿌리로 (The Korean Pharmacopeia Tenth Edition, 2010), 최근 한의학에서널리상용되는대표거담제천남성에대한각종약 리작용및효과에대한연구가진행되고있으며 ATP 효소활성 화효과, 혈관긴장성두통, 삼차신경에의한안면신경통등기타 의통증에대한진통, 진정효과가실험적으로입증되고있다 (Jeon, 2000). 천남성의주성분은 triterpenoid, saponin, amino acid, benzoic acid 등다양한성분이밝혀져있으며, 이중 d- mannitol 은항암활성이있다고증명되었다 (Jing et al., 1996). * 교신저자 : ezhani@kiom.re.kr Tel 천남성은강한자극이있어서생것을입에넣으면입안이부르트고목안이아리고, anemonin 계등의유독성분이함유되어있어서허약한사람이나임신부에게는쓰는것을조심해야하는생약으로국내에서보고된사례에는음독에의한구강과인두의충혈과통증부종에대한사례가있으며 (Hong et al., 2003), 독성작용에의한촉각과미각의소실, 타액분비의소실, 점막의궤양, 인두부의부종과심한경우목소리의소실등이알려져있다 (Jang, 2003; Huaug, 1999; Bensky, 1986). 한약및생약에대한수요가증가함에따라다양한한약과생약을원료로사용하는민간요법에서부터기능성식품및천연물관련의약품등까지무분별한사용되어지고있고 (Oh et al., 2010; Choi et al., 2014), 이에따라안전성이확보되지않은식품및의약품이오 남용되고있어독성및부작용이늘어나게됨으로써철저한규제및관리가요구되고있으며특히, 천연물에대한과학적검증을통한안전성확보및실험적근거를통해이를입증해야하는상황이요구되고있다 (Hwang et al., 2004; c 본학회지의저작권은 ( 사 ) 한국자원식물학회지에있으며, 이의무단전재나복제를금합니다 This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

2 韓資植誌 Korean J. Plant Res. 28(4) : 371~381(2015) Yoo et al., 2007). 근래에한약자체단미제의독성평가가활발히진행되고있으며, 평가로는금은화 (Yoo et al., 2010), 반하 (Lee et al., 2003), 죽력 (Chang and Hong, 1985), 황금 (Lee et al., 2013), 포공영 (Gu et al., 2011), 황기 (Park et al., 2013) 등이보고되어왔다. 본연구에서는장기간복용되었을경우있을수있는독성에대한안전성평가를실시하기위해천남성추출물을 Sprague- Dawley 랫드에게 28일동안반복경구투여하여일반증상및사망동물을관찰하였으며, 체중변화, 안검사, 요검사, 혈액학적및혈액생화학적검사, 혈액응고, 장기중량등을실시하여독성에대한기초자료를제공하고자독성의질적, 양적인평가는식품의약품안전처독성시험관리기준 (Korea food and drug administration, 2005 (KFDA)) 과 OECD 검정기준 (Organization for Economic Co-operation and Development, 2011) 에준하여실시하였다. 재료및방법재료약재본실험에서사용된천남성 (Arisaema Rhizome) 은대한약전에규정된, 주피를완전히제거한덩이뿌리상태를약업사 ( 광명당제약, K ) 에서구입하였으며, 현미경으로관능검사를실시한뒤시료로선정하여본실험에사용하였다. 천남성추출천남성은경구투여시열수추출물에의해복용되므로, 본실험은열수추출하였다. 선정된천남성 800 g을 3,000~5,000 ml의음용수에넣어 2시간동안끓여, 그후남은양 (1,600 ml ) 속에천남성 800 g을녹인뒤, 5 g/10 ml로맞추어본시험의초고용량을 5,000 mg / kg /day 로하고중, 저용량을음용수로단계희석하였다. 마우스경구단회투여독성실험실험동물및사양관리암수각 20마리의 SPF 마우스 (6-wk old upon receipt, Sparague Dawley R TM SDR TM 랫드, ( 주 ) 대한바이오링트를 5 일간의순화과정을거쳐실험에사용하였으며, 순화과정및실험전기간동안온도 23 ± 3, 상대습도 50 ± 20% 온 습도범위를유지하였다. 순화, 투여및관찰기간중 Polysulfone 사 육상자 (260 W 420 L 150H mm ) 에 2~3 마리씩수용하여사육 하였고, 명암 cycle 은형광등조명 12 시간주기로조절하였으며 ( 조도 150~300Lux), 사료 (Zeigler Bros., Inc. USA) 와음수는 자유롭게공급하였다. 모든실험동물은투여일및최종부검일 17 시간전절식을실시하였으며, 부검당일 Isoflurane 으로흡 입마취하여복대동맥에서혈액학적검사및혈액생화학적검 사를위한채혈을실시하였다. 군분리및약물의투여 실험동물은군당 5 마리씩 Table 1 에기록한 8 그룹으로구분 하여식품의약품안전처기준 (Korea food and drug administration, 2005 (KFDA)) 과 OECD 실험기준 (Organization for Economic Co-Operation and Development, 2001) 에의거하 여, 일반적인반복투여독성시험한계용량인 1,000 mg / kg /day 보다 5 배높은 5,000 mg / kg /day 최고용량으로설정하였다. 그 리고 5,000 mg / kg /day 이상의용량에서는시험물질자체의독 성보다과용량에의한물리적및생리적부작용에영향을주기 때문에이이상더투여는고려되지않아, 설치류최고한계투여 용량인 5,000 mg / kg /day 을최고용량으로하고공비 2 를두어 2,500, 1,250 mg / kg /day 을중, 저용량군으로군을설정하였다. 또한암수각각에대한매체대조군을추가하였다. 모든투여군 에서는최근체중측정일의체중을기준으로투여액량을 10 ml / kg 으로산출하여천남성추출물을 overnight 절식후경구투여용 존데를장착한주사관을이용하여위내에직접투여하였다. 투 여횟수는 1 회 / 일, 7 일 / 주, 4 주간투여하였으며, 투여는 12:00 Table 1. Experimental design used in single dose toxicity test Group Sex NO. of animals Dosage ( ml / kg /day) Total Dose ( ml / kg ) G1M z Male G2M Male ,250 G3M Male ,500 G4M Male ,000 G1F z Female G2F Female ,250 G3F Female ,500 G4F Female ,000 z M: Male, F: Female. Vehicle control: distilled water 20 ml / kg as vehicle in this study. All test articles in vehicle were once orally dosed in a volume of 10 ml / kg. Dissolved in distilled water

3 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 이전에완료하였다. 임상증상의관찰모든실험군에대해서 1일 1회 functional observational battery test (Irwin, 1968; Dourish, 1987) 를기초하여일반증상의종류, 발현일및증상의정도를관찰하였으며, 1일 2회사망및빈사동물을관찰하였다. 관찰은투여 0일부터 28일까지 4주간실시하여개체별로기록하였다. 체중의측정모든실험동물의체중을투여개시일에측정하고, 이후에는주 1회, 부검전일및부검일에측정하였다. 부검일체중은절식을실기하였으므로, 체중평가에서제외하고부검전일의체중은하룻밤절식시킨후의체중을평가하였다. 사료섭취량산정투여개시전의사료섭취량은군분리일부터투여개시일까지 1일간섭취량을측정하였다. 투여기간에는 7일간의섭취량을측정하여 1일평균섭취량을산출하였다. 투여 4주째에는 6일간의섭취량을측정하여마리당평균섭취량 (g/rat/day) 으로 1일의평균섭취량을산출하였다. 안검사동물도입시모든동물에대하여육안으로눈의외관을관찰하였다. 투여마지막주에는모든동물을육안으로눈의외관관찰한후, 각군당 5마리에대해서는안구에산동제 (Ocutropic ophthalmic drops, 로트번호 : , Samilpharma. Co., Ltd.) 를점적하여동공확장을유도한다음, 안저사진기 (Genesis, Gowa Co. Ltd., Japan) 로전안부, 중간투광체및안저를관찰하였다. 요검사각군당 5마리에대하여투여 4주에요검사를실시하였다. 동물을대사케이지에수용하여배설후약 3~4 시간이내채뇨한신선뇨및배설후약 24시간축뇨 1 ml를취하여일반검사, 요침사검사를수행하였다. 24 시간동안계속채집한요로요총량을측정하였다. 일반검사약 0.3 ml의요를요검사용시험지 (Multistix 10SG, SIMENS, USA) 에묻힌후, 요자동분석기 (CliniTex Advantus TM, SIEMES, USA) 를이용하여 GLU (glucose, g/ dl ), BIL (bilirubin, mg / dl ), KET (ketone body, mg / dl ), LEU (leukocyte, Leu/ μl ), OB (occult blood, Ery/ μl ), PRO (protein, mg / dl ), URO (urobilinogen, umol/l), NIT (nitrite), SG (specific gravity), ph, Color 를측정하였다. 요침사검사일반검사후남은요를 1,500 rpm 으로 5분간원심분리 (MF80, Hanil, Korea) 한후, 그침전물을 S.M (sternheimermalbin) 법으로염색하여 RBC (red blood cell), WBC (white blood cell), Epithelial cell 및 Casts 를관찰하였다. 혈액학적검사부검시채혈한혈액일부를항응고제인 EDTA 2K 가들어있는 CBC bottle (Vacutainer 3 ml, SEKISUI, JAPAN) 에주입한후자동혈액분석기 (ADVIA 2120, SIEMENS, USA) 로일반혈액학적검사와백혈구감별계수를나누어측정하였다. 일반혈액학적검사 WBC (white blood cell count), RBC (red blood cell count), HGB (hemoglobin conc.), HCT (hematocrit), MCV (mean corpuscular volume), MCH (mean corpuscular hemoglobin), MCHC (mean corpuscular Hb conc.), RDW (red cell distribution width), HDW (hb conc. distribution width), PLT (platelet), MPV (mean platelet volume), RET (reticulocyte). 백혈구감별계수 NEU (neutrophil), LYM (lymphocyte), MONO (monocyte), EOS (eosinophil), BASO (basophil), LUC (large unstained cells). 혈액생화학적검사혈액일부를 clot activator 가들어있는 vacutainer tube (SEKISUI, Japan) 에주입하고 15~20 분간상온에방치하여응고시켰다. 이후 3,000 rpm 으로 10분간원심분리 (MF80, Hanil, Korea) 하여얻은혈청으로혈액상화학분석기 (KONELAB 20XT, Thermo, USA) 를활용하여 AST (aspartate aminotransferase), ALT (alanine aminotransferase), ALP (alkaline phosphatase), BUN (blood urea nitrogen), CRE (creatinine), GLU (glucose),

4 韓資植誌 Korean J. Plant Res. 28(4) : 371~381(2015) CHO (total cholesterol), PRO (total protein), CP K(creatine phosphokinase), ALB (albumin), BIL (total bilirubin), TG (triglyceride), IP (inorganic phosphorus), A/G ratio (albumin/ globulin ratio) 의항목을측정하였다. 또한 Na+ (sodium ion), K+ (potassium ion), Cl (chloride ion) 의항목을전해질분석기 (744 Na + /K + /Cl - / Analyzer, SIEMES, USA) 로측정하였다. 장기중량측정육안부검소견을관찰기록한후난소 (ovary), 부신 (adrenal gland), 뇌하수체 (pituitary), 가슴샘 (thymus), 전립샘 (prostate), 고환 (testis), 부고환 (epididymis), 비장 (spleen), 신장 (kidney), 심장 (heart), 폐 (lung), 뇌 (brain) 및간장 (liver) 을적출한후전자저울을이용하여중량을측정하였다. 양측성장기는양측장기를각각측정하였다. 중량측정장기에대하여는부검시체중에대한상대중량을산출하였다. 조직병리학적관찰난소 (ovary), 부신 (adrenal gland), 뇌하수체 (pituitary), 가슴샘 (thymus), 전립샘 (prostate), 고환 (testis), 부고환 (epididymis), 비장 (spleen), 신장 (kidney), 심장 (heart), 폐 (lung), 뇌 (brain) 및간장 (liver) 등의주요장기중량을측정한후일부조직을 10% 중성완충포르말린용액에고정하되, 고환과부고환은 Bouin s 용액에고정하였다. 통계처리및결과판정대조군과투여군간의평균비교에는연속적인자료의분석으로모수적인비중비교 (parametric multiple comparison procedures) 과불연속적인자료의분석으로비모수적인다중비교 (non-parametric multiple comparison procedures) 를사용하였다. 모수적인검사인체중, 사료섭취량, 혈액학적검사, 혈액생화학적검사, 장기중량에대하여일원배치분산분석 (one-way analysis of variance test) 로평균치에대한유의성를검정하였다. 유의성이있으면 Levene test 로등분산성을검정하여등분산일경우에는 Duncan multiple range test, 이분산일경우에는 Dunnett s T-test 를이용하였다. 비모수검사인요검사결과자료는아래표같이척도변환을실시한후, Kruskal-Wallis s H-test 를실시한후, 유의성이있으면 Mann-Whitney U-test 를적용하여대조군과유의성을확인하였다. 요색조는 Fisher s exact test 를적용하였다. 통계학적분석은상용으로널리사용되고있는통계패키지인 SPSS 14.0 을이용하였다. 결과 마우스경구단회투여독성실험일반증상및사망률천남성추출물투여와관련있는사망례는실험기간중대조 Table 2. Mortalities observed in female and male mice of single dose toxicity test z Group ID y Day after dosing 0 x Total w G1M v /5 (0%) G2M /5 (0%) G3M /5 (0%) G4M /5 (0%) G1F v /5 (0%) G2F /5 (0%) G3F /5 (0%) G4F /5 (0%) z Number of died animals. y Group ID was listed in Table 1. x Dosing day. w Total mortalities during 28 days of observation periods died animals/total observed animals (n=5). v M: Male, F: Female

5 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 군을포함한모든시험군에서사망동물은없었으며, 투여중일 반증상관찰결과이상소견은없었다. 모든실험동물 (5/5; 100%) 에서최종부검을실시하였다 (Table 2). 안과학적검사소견 군분리시외안검사와투여마지막주에수행한안저검사결과, 암수모든시험군에서어떠한증상도관찰되지않았다 (Table 4). 체중및증체량의변화체중변화 (body weight change) 및증체량 (body weight gains) 을관찰한결과, 모든시험물질투여군증제량이부형제대조군에비해다소증가경향을보였으나, 천남성추출물투여시암수모두에서대조군과비교하여유의성있는체중변화는없었다 (Table 3). 요검사소견요검사결과 (Table 5-1, 5-2, 5-3), 암수모든시험군에서 0~5,000 mg / kg투여군에서천남성추출액에의한영향으로판단되는결과는없었다. 그외, 뇨의투명도 (transparency) 의변화, 뇨단백 (protein), 케톤체 (ketone body) 가관찰되었으나, 모두미량의검출로서용량의존성이없거나신장의형태학적변화가관찰되지않았으므로독성학적의미는없다고사료된다. Table 3. Body weight gains in female and male mice of single dose toxicity test z Group ID y Interval Day 0 x ~7 Day 8~14 Day 15~21 Day 22~28 Gains w N G1M v ± ± ± ± ± G2M ± ± ± ± ± G3M ± ± ± ± ± G4M ± ± ± ± ± G1F v ± ± ± ± ± G2F ± ± ± ± ± G3F ± ± ± ± ± G4F ± ± ± ± ± z Values are expressed as mean ± S.D. of four mice. y Group ID was listed in Table 1. x Day of dosing after overnight fasted. w Weight gains in the main groups are body weight difference between week 4 and 1stdosed day. v M: Male, F: Female. Table 4. Ophthalmic findings in female and male after treatment Group ID z Sites Observed signs Left eye Right eye No observed signs No observed signs G1M y 5 / 5 5 / 5 5 / 5 5 / 5 G2M 5 / 5 5 / 5 5 / 5 5 / 5 G3M 5 / 5 5 / 5 5 / 5 5 / 5 G4M 5 / 5 5 / 5 5 / 5 5 / 5 G1F y 5 / 5 5 / 5 5 / 5 5 / 5 G2F 5 / 5 5 / 5 5 / 5 5 / 5 G3F 5 / 5 5 / 5 5 / 5 5 / 5 G4F 5 / 5 5 / 5 5 / 5 5 / 5 z Values are expressed as mean ± S.D. of four mice. y M: Male, F: Female

6 韓資植誌 Korean J. Plant Res. 28(4) : 371~381(2015) Table 5-1. Urinalysis of male and female rats Tests z Result Grade Male Female G1M y G2M G3M G4M G1F y G2F G3F G4F 0 1,250 2,500 5, ,250 2,500 5,000 GLU BIL KET / SG ph PRO / URO NIT OB / LEU / z The unit of urobilinogen is ehrlich unit / dl. y M: Male, F: Female. GLU: glucose BIL: bilirubin KET: ketone body SG: specific gravity PRO: protein URO: urobilinogen. NIT: nitrite OB: occult blood LEU: leukocyte. 요검사결과, GLU, BIL, URO 에서는모든군에서음성으로관찰되었으며, SG, ph, PRO, LEU 의약물주입에따른용량의존적인변화는관찰되지않았다. KET 의경우수컷 1,250 과 5,000 ( mg / kg /day) 에서반양성이관찰되었으나통계적유의성은관찰되지않았다. OB는수컷에서 NIT, LEV 는암컷군에서감소하는경향을보였다. 혈액학적검사부검시체혈한혈액으로혈액학적검사를실시한결과 (Table

7 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 Table 5-2. Urine sediments of male and female rats Urine sediments Tests Result Severity Male Female G1M z G2M G3M G4M G1F z G2F G3F G4F 0 1,250 2,500 5, ,500 3,000 5,000 RBC / WBC / EPITHELIAL CELL CASTS / No. of animals z M: Male, F: Female. Table 5-3. Urine volume and color of male and female rats Group ID z Urine volume and color of rats Tests Volume ( ml ) Color-yellow No. of animals G1My 10.4 ± G2M 12.0 ± G3M 11.8 ± G4M 11.8 ± G1F y 8.8 ± G2F 8.4 ± G3F 9.4 ± G4F 8.0 ± z Values are expressed as mean ± S.D. of four mice. y M: Male, F: Female. 6), 호중구 (NEU), 단구 (MONO) 및호산구 (EOS) 는암컷 HCT 수치가고용량군에서유의한감소를 (p<0.05) 나타냈으나정상범위 (historical data) 내변화이며, PLT 수치도중용량군에서유의한감소가 (p<0.05) 나타냈으나정상범위내변화로판단되어비시험물질 -유래변화로추정된다. 래변화로추정된다. 또한 PRO, A/G 수치도저용량군에서각각유의한증가및감소가 (p<0.05) 나타냈으나용량의존성이없고정상범위내변화로판단되어비시험물질 -유래변화로추정된다. 혈액응고검사결과, 대조군과비교하여특이변화가관찰되지않았다 (Table 8). 혈액생화학적검사생화학적검사결과 (Table 7), 수컷의 AST 수치가대조군과비교하여중용량군에서유의한감소가 (p<0.05) 나타냈으나용량의존성이없고정상범위내변화로판단되어비시험물질 -유 장기중량검사 장기중량의측정결과 (Table 9), 대조군과비교하여특이변 화가관찰되지않았다

8 韓資植誌 Korean J. Plant Res. 28(4) : 371~381(2015) Table 6. Hematological values of male and female rats Hematological values Tests Units G1M z (0) G2M (1,500) G3M (3,000) G4M (5,000) G1F z (0) G2F (1,500) G3F (3,000) G4F (5,000) WBC 103/μL 2.75 ± ± ± ± ± ± ± ± RBC 106/μL 7.96 ± ± ± ± ± ± ± ± HGB g/dl ± ± ± ± ± ± ± ± HCT % ± ± ± ± ± ± ± ± MCV fl ± ± ± ± ± ± ± ± MCH pg ± ± ± ± ± ± ± ± MCHC g/dl ± ± ± ± ± ± ± ± RDW % ± ± ± ± ± ± ± ± HDW g/dl 2.77 ± ± ± ± ± ± ± ± PLT 103/μL ± ± ± ± ± ± ± ± MPV fl 8.56 ± ± ± ± ± ± ± ± RET % 1.55 ± ± ± ± ± ± ± ± NEU % ± ± ± ± ± ± ± ± /μL 0.44 ± ± ± ± ± ± ± ± LYM % ± ± ± ± ± ± ± ± /μL 2.20 ± ± ± ± ± ± ± ± MONO % 1.74 ± ± ± ± ± ± ± ± /μL 0.04 ± ± ± ± ± ± ± ± EOS BASO LUC % 1.74 ± ± ± ± ± ± ± ± /μL 0.05 ± ± ± ± ± ± ± ± % 0.14 ± ± ± ± ± ± ± ± /μL 0.00 ± ± ± ± ± ± ± ± % 0.50 ± ± ± ± ± ± ± ± /μL 0.01 ± ± ± ± ± ± ± ± No. of animals z M: Male, F: Female 부검소견부검결과에서암수모두에서가슴샘의발적 (redness) 이, 암컷에서자궁내맑은액체저류 (retention of clear fluid) 가관찰되었다. 하지만시험물질의영향이아닌일반 Rat 에서흔히나타나는증상으로비시험물질 -유래변화로추정된다. 고찰 본시험은천남성추출물의반복투여에따른안전성을평가하고자 6주령 Sprague-Dawley 계랫드에시험물질을 1,250, 2,500 및 5,000 mg / kg의용량으로군당암수각 5 마리에 28일간경구투여하였다. 독성지표를위해일반증상관찰, 체중측정, 사료섭취량측정, 안검사, 요검사, 혈액학적및혈액생화학적검사, 부검시장기의중량측정및육안적검사등이수행되었다. 시험물질에대한영향및이에대한판단은본시험의판단기준에따라다음과같이확인되었다. 1. 투여기간중사망동물은관찰되지않았으며, 일반증상관찰소견에서도시험물질의영향으로보이는특이증상은없었다. 2. 관찰기간동안체중변화 (body weight changes), 증체량 (weight gains) 및사료섭취량에서시험물질에의한변화는관찰되지않았다. 3. 요검사에서수컷 OB 수치가중용량및고용량군에서유의한감소를 (p<0.05) 보였으나정상범위 (Historical data) 내변화이며, 대조군및저용량군에서만관찰되어용량의존성이없어비시험물질-유래변화로추정된다. 4. 혈액학적검사에서암컷 HCT 수

9 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 Table 7. Serum biochemical values of male and female rats Tests Units G1M z (0) G2M (1,250) G3M (2,500) G4M (5,000) G1F z (0) G2F (1,500) G3F (3,000) G4F (5,000) ALB g/dl 3.19 ± ± ± ± ± ± ± ± ALP U/l ± ± ± ± ± ± ± ± ALT U/l ± ± ± ± ± ± ± ± AST U/l ± ± ± ± ± ± ± ± BIL mg /dl 0.10 ± ± ± ± ± ± ± ± BUN mg /dl ± ± ± ± ± ± ± ± CHO mg /dl ± ± ± ± ± ± ± ± CPK U/l ± ± ± ± ± ± ± ± CRE mg /dl 0.42 ± ± ± ± ± ± ± ± GLU mg /dl ± ± ± ± ± ± ± ± IP mg /dl 5.68 ± ± ± ± ± ± ± ± PRO g/dl 5.53 ± ± ± ± ± ± ± ± TG mg /dl ± ± ± ± ± ± ± ± A/G ratio 1.37 ± ± ± ± ± ± ± ± Na+ mg /dl ± ± ± ± ± ± ± ± K+ mmol/l 4.00 ± ± ± ± ± ± ± ± Cl- mmol/l ± ± ± ± ± ± ± ± z M: Male, F: Female. Table 8. Plasma coagulation values of male and female rats Tests Plasma coagulation values G1M z (0) G2M (1,250) G3M (2,500) G4M (5,000) G1F z (0) G2F (1,250) G3F (2,500) G4F (5,000) PT (sec) 9.55 ± ± ± ± ± ± ± ± APTT (sec) ± ± ± ± ± ± ± ± No. of animals z M: Male, F: Female 치가고용량군에서감소가 (p<0.05) 관찰되었으나정상범위 (historical data) 내변화이며, PLT 수치도중용량군에서유의한감소가 (p<0.05) 관찰되었으나정상범위내변화로판단되어비시험물질 -유래변화로추정된다. 5. 생화학적검사결과, 수컷의 AST 수치가대조군과비교하여중용량군에서유의한감소가 (p<0.05) 관찰되었으나용량의존성이없고정상범위내변화로판단되어비시험물질 -유래변화로추정된다. 또한 PRO, A/G 수치도저용량군에서각각유의한증가및감소가 (p<0.05) 관찰되었으나용량의존성이없고정상범위내변화로판단되어비시험물질 -유래변화로추정된다. 적요 본연구는민속식물로많이사용되는천남성 (Arisaema amurense f. serratum (Nakai) Kitag) 추출물의반복투여에따른안전성을평가하기위하여 6 주령 Sprague-Dawley 계 Rat 를이용한실험이다. 각암수 5 마리에게 28일간 1,250, 2,500 그리고 5,000 mg / kg /day 의농도별열수추출물을경구투여하여독성지표를위해일반증상관찰, 체중측정, 안검사, 요검사, 혈액학적및혈액생화학적검사등을측정하였다. 본시험을통해암수모두에게 1,250, 2,500 그리고 5,000 mg / kg의모든투여농도에서시험물질에의한변화관찰되지않아비시험물질 -유래변화가확인되었다. 따라서시험의독성판정기준에따라암수모

10 韓資植誌 Korean J. Plant Res. 28(4) : 371~381(2015) Table 9. Relative organ weights of male and female rats (% body weight) Organ weight (g) Absolute of organ weights (g) G1M z (0) G2M (1,500) G3M (3,000) G4M (5,000) G1F z (0) G2F (1,500) G3F (3,000) G4F (5,000) Adrenal (L) ± ± ± ± ± ± ± ± Adrenal (R) ± ± ± ± ± ± ± ± Pituitary ± ± ± ± ± ± ± ± Testis (L) ± ± ± ± Ovary (L) ± ± ± ± Testis (R) ± ± ± ± Ovary (R) ± ± ± ± Epididymis (L) ± ± ± ± Epididymis (R) ± ± ± ± Thymus ± ± ± ± ± ± ± ± Prostate ± ± ± ± spleen ± ± ± ± ± ± ± ± kidney (L) ± ± ± ± ± ± ± ± kidney (R) ± ± ± ± ± ± ± ± Heart ± ± ± ± ± ± ± ± Lung ± ± ± ± ± ± ± ± Brain ± ± ± ± ± ± ± ± Liver ± ± ± ± ± ± ± ± No. of animals z M: Male, F: Female. 두 NOEL ( 최대무영향용량 ) 은 5,000 mg / kg /day 로추정된다. 이상의결과를바탕으로추가적임상시험을진행하여천남성안전성검사를검증해야한다. 사사 본연구는국가과학기술연구회 YS 인건비 ( ) 와한국한의학연구원민간요법발굴 보존및 DB구축을통한지식자원화 (K15060) 의지원으로이루어졌다. References Bensky, D. and A. Gamble Chinese Herbal Medicine - Revised Edition. Eastland Press, Seattle, WA (USA). Chang, I.K and N.D. Hong Experimental studies on the acute toxicity and the efficacy of phyllostachys juice. Korean J. Orient. Int. Med. 2: Choi, H.K., H.S. Roh, J.Y. Jeong and H.Y. Ha Acute oral toxicity of Atractylodes macrocepala KOIDZ. Korean J. Plant Res. 27(1):11-21 (in Korean). Dourish, C.T Effects of drugs on spontaneous motor activity: In Greenshaw, A.J. and C.T. Dourish (eds.), Experimental Psychopharmacology. Humana Press. Clifton, New York (USA). pp Gu, J.H., S.R. Kim, J.W. Lee, M.Y. Park, H.Y. Choi and J.D. Kim Mouse single oral dose toxicity test of Taraxaci Herba aqueous extracts. J. of Oriental Medical Pathology 25(4): Hong, M.K., J.O. Park and S.J. Wang A case of toxicity caused by Arisaematis Rhizoma ingestion. J. The Korean Society of Clinical Toxicology 1(1): Huaug, K.C The Pharmacology of Chinese Herb II. CRC press, Boca Raton (USA). Hwang, S.Y., W. Kwon, H.Y. Chai, Y.M. Cho, N.J. Lee, J.M. Ryu, J.S, Sin, T.M. Kim, J.H. Cho, E.J.Kim, J.H. Pakr, J.K. Kang and Y.B. Kim Four-week repeated-dose toxicity study on Mori Raddiceis cortex. The Korean Journal of Laboratory Animal Sci. 20(3): Irwin, S Comprehensive observational assessment: Ia. A

11 천남성추출물의 Sprague-Dawley 랫드를이용한 28 일반복경구투여 DRF 독성시험 systemic, quantitative procedure for assessing the behavioral and physiological state of the mouse. Psychopharmacology 13: Jang, I.Y Treaties on Asian Herbal Medicines I. Natural Products Research Institute, Seoul National University Press, Korea. pp Jeon, H Effect of Arisaematis Rhizoma on the proliferation of transplanted - L1210 cells in Mice. Kor. J. Herbology 15(2):9-16. Jing, J.H., C.O. Lee, Y.C. Kim and S.S. Kang New bioctive cerebrosides from arisaema amurense. J. Nat. Prod. 59: Korea Food and Drug Administration Testing guidelines for safety evaluation of drugs, notification No Korea Food and Drug Administration, Seoul, Korea. Lee, L.E., H.J. Kim, E.K. Choi, H.Y. Chai, Y.W. Yun, D.J. Kim and S.Y. Nam Four-week repeated-dose toxicity study on Pinellia Extract. Korean J. Lab. Anim. Sci. 19: Oh, T.W., H.S. Bae, C.H. Yoon and Y.K. Park Thirteen- week repeated-dose oral taxicity study of the modified Wenpitang-Hab-Wulingsan (WHWR) in sprague-dawly rats. Korean Journal of Herbol. 25(3): Park, Y.C., J.S. Lee, D.Y. Kim, H.Y. Son, J.W. Lee, Y.S. Chei, K.K. Kim, C.Y. Yu, I.M. Chung, M.H. Im, K.J. Lee, R.N. Choi, H.S. Shim and J.D. Lim A 90 day repeated dose-oral toxicity study of extracts from Astragalus membranaceus-aboveground parts in rats. Korean J. Medicinal Crop Sci. 21(6): Yoo, T.W., B.I. Kim, J.B. Kim, D.J. Kim, J.W. Kim, S.K. Baik, K.S. Kim and G.J. Cheon Survey for the actual condition of drug medication and development of health care cost associated with toxic liver injury in Korean: A multicenter study for the detection and the development of nationwide reporting system of toxic. Clinical and Molecular Hepatology 13: Yoo, H.J., M.Y. Park, H.Y. Choi and J.D. Kim Mouse single oral dose toxicity test of Lonicerae Flos aqueous extracts. Korean J. Oreiental Med. 31(3): (Received 18 September 2014 ; Revised 14 April 2015 ; Accepted 8 June 2015)

025~038-오공약침의

025~038-오공약침의 25 DOI : 10.3831/KPI.2008.11.4.025 A Study on Subchronic Toxicity Test and Method of Increasing Output of Scolopendrid Pharmacopuncture Kim Sung-chul *Dept. of Acupuncture & Moxibustion, Gwang-Ju Oriental