독활의 복합 유산균 발효 추출액의 마우스에 대한 단회경구투여 독성시험 Mouse Single Oral Dose Toxicity Test of Lactobacillus-fermented Araliae Continentalis Radix Aqueous Extracts (fACR)원문보기
본 실험에서는 유산균발효 독활의 마우스 단회 경구 투여 독성 자료를 얻기 위해 식품의약품안전청 고시 제 2013-121 “의약품 등의 독성시험 기준”에 의거하여, 설치류 투여 한계 용량인 2,000 mg/kg을 최고 투여군을 설정하고 공비 2로 1,000 및 500 mg/kg 투여군을 중간 및 저용량 투여군으로 설정하여 실험을 실시하였으며, 그 결과는 독활 열수 추출물 2,000 mg/kg 암수 투여군 및 암수 매체 대조군과 비교 평가 하였다. 본 실험의 결과, 설치류 투여한계 용량인 2,000 mg/kg 투여군까지, 유산균발효 독활 열수 추출물 투여와 관련된 사망례, 임상증상, 체중, 장기중량, 육안부검 및 조직병리학적 소견이 인정되지 않았다. 따라서 유산균발효 독활 열수 추출물의 마우스에 대한 단회 경구 투여 반수 치사량 및 개략적 치사량은 암수 각각 2,000 mg/kg이상으로 산출되었으며, 특정 임상증상 및 표적 장기 역시 없는 것으로 판단되어, 유산균발효 독활은 매우 안전한 물질로 판단된다. 또한 독활 열수 추출물 2,000 mg/kg 투여와 관련된 사망례, 임상 증상, 체중, 장기중량, 육안 및 조직병리학적 변화 역시 인정되지 않았다. 이러한 결과는 독활의 활용도를 증대시키는 기초 자료가 될 것으로 사료된다.
본 실험에서는 유산균발효 독활의 마우스 단회 경구 투여 독성 자료를 얻기 위해 식품의약품안전청 고시 제 2013-121 “의약품 등의 독성시험 기준”에 의거하여, 설치류 투여 한계 용량인 2,000 mg/kg을 최고 투여군을 설정하고 공비 2로 1,000 및 500 mg/kg 투여군을 중간 및 저용량 투여군으로 설정하여 실험을 실시하였으며, 그 결과는 독활 열수 추출물 2,000 mg/kg 암수 투여군 및 암수 매체 대조군과 비교 평가 하였다. 본 실험의 결과, 설치류 투여한계 용량인 2,000 mg/kg 투여군까지, 유산균발효 독활 열수 추출물 투여와 관련된 사망례, 임상증상, 체중, 장기중량, 육안부검 및 조직병리학적 소견이 인정되지 않았다. 따라서 유산균발효 독활 열수 추출물의 마우스에 대한 단회 경구 투여 반수 치사량 및 개략적 치사량은 암수 각각 2,000 mg/kg이상으로 산출되었으며, 특정 임상증상 및 표적 장기 역시 없는 것으로 판단되어, 유산균발효 독활은 매우 안전한 물질로 판단된다. 또한 독활 열수 추출물 2,000 mg/kg 투여와 관련된 사망례, 임상 증상, 체중, 장기중량, 육안 및 조직병리학적 변화 역시 인정되지 않았다. 이러한 결과는 독활의 활용도를 증대시키는 기초 자료가 될 것으로 사료된다.
The objective of this study was to obtain acute (single) oral dose toxicity information on Lactobacillus-fermented Araliae Continentalis Radix aqueous extracts (fACR) in female and male ICR mice, as compared with Araliae Continentalis Radix aqueous extracts (ACR). After administering a single oral d...
The objective of this study was to obtain acute (single) oral dose toxicity information on Lactobacillus-fermented Araliae Continentalis Radix aqueous extracts (fACR) in female and male ICR mice, as compared with Araliae Continentalis Radix aqueous extracts (ACR). After administering a single oral dose of fACR, no treatment-related mortalities were observed within 14 days after the end of treatment up to 2,000 mg/kg, the maximum dosage for rodents of both sexes; moreover, no fACR treatment-related changes in the body and organ weights, clinical signs, necropsy, and histopathological findings were detected in this experiment. In addition, no ACR 2,000 mg/kg treatment-related mortalities, clinical signs, body and organ weights, or gross and histopathological findings were observed, as compared with equal genders of vehicle control. The results obtained in this study suggest that fACR is non-toxic in mice and is, therefore, likely to be safe for clinical use. The LD50 and approximate LD in female mice and male mice, respectively, were considered after a single oral dose of fACR over 2,000 mg/kg, the maximum dosage for rodents. In addition, no specific targets or clinical signs were detected in the present study. ACR 2,000 mg/kg-treated mice also did not show any treatment-related mortalities, clinical signs, changes to body and organ weights, or gross and histopathological findings, as compared with equal genders of vehicle control.
The objective of this study was to obtain acute (single) oral dose toxicity information on Lactobacillus-fermented Araliae Continentalis Radix aqueous extracts (fACR) in female and male ICR mice, as compared with Araliae Continentalis Radix aqueous extracts (ACR). After administering a single oral dose of fACR, no treatment-related mortalities were observed within 14 days after the end of treatment up to 2,000 mg/kg, the maximum dosage for rodents of both sexes; moreover, no fACR treatment-related changes in the body and organ weights, clinical signs, necropsy, and histopathological findings were detected in this experiment. In addition, no ACR 2,000 mg/kg treatment-related mortalities, clinical signs, body and organ weights, or gross and histopathological findings were observed, as compared with equal genders of vehicle control. The results obtained in this study suggest that fACR is non-toxic in mice and is, therefore, likely to be safe for clinical use. The LD50 and approximate LD in female mice and male mice, respectively, were considered after a single oral dose of fACR over 2,000 mg/kg, the maximum dosage for rodents. In addition, no specific targets or clinical signs were detected in the present study. ACR 2,000 mg/kg-treated mice also did not show any treatment-related mortalities, clinical signs, changes to body and organ weights, or gross and histopathological findings, as compared with equal genders of vehicle control.
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문제 정의
The objective of this study was to obtain acute (single) oral dose toxicity information of Lactobacillus fermented Araliae Continentalis Radix aqueous extracts (fACR) in fe- male and male ICR mice as compared with those ACR. Because there are no available toxicological data after oral treatment in female and male mice of ACR or fACR, the highest dosage used in the present study were selected as 2, 000 ㎎/㎏ in a volume of 20 ml of distilled water - the limited dosages of rodents [6], and 1,000 and 500 ㎎/㎏ were selected as middle and lower dosage groups according to KFDA Guidelines [16].
제안 방법
Body weights were measured at the day of dosing (Day 0) immediately before treatment, 1, 2, 7, 13 and 14 days after treatment using automatic electronic balance (Precisa Instrument, Zuerich, Switzerland). In addition, to reduce in- dividual body weight differences of animals at treatment, body weight gains during Day 0 ~ Day 7, Day 7 ~ Day 13 and Day 0 ~ Day 14 was also calculated based on measured body weight at each point.
Each of twenty-seven female and male specific patho- gen-free VAF Outbred CrljOri:CD1 [ICR] mice (6-wk old upon receipt, OrientBio, Sungnam, Korea; ANNEX I and II) were used after acclimatization for 9 days. Animals were allocated five per polycarbonate cage in a temperature (20-25 ℃) and humidity (40-45%) controlled room.
In addition each female and male vehicle controls were added with ACR 2,000 ㎎/㎏. In or- der to investigate the toxicity and identify target organs, fACR were once orally administered to female and male ICR mice at dose levels of 2,000, 1,000, 500 and 0 (vehicle control) ㎎/㎏ (body weights) in a volume of 20 ml/kg, dissolved in distilled water, and the mortality and changes on the body weight and clinical signs were monitored during 14 days after treatment with gross observation, changes on the organ weights and histopathology of principle organs based on the recommendation of KFDA Guidelines [16], as compared with those of vehicle control and ACR 2,000 ㎎/㎏ treated mice.
In addition each female and male vehicle controls were added with ACR 2,000 ㎎/㎏. In or- der to investigate the toxicity and identify target organs, fACR were once orally administered to female and male ICR mice at dose levels of 2,000, 1,000, 500 and 0 (vehicle control) ㎎/㎏ (body weights) in a volume of 20 ml/kg, dissolved in distilled water, and the mortality and changes on the body weight and clinical signs were monitored during 14 days after treatment with gross observation, changes on the organ weights and histopathology of principle organs based on the recommendation of KFDA Guidelines [16], as compared with those of vehicle control and ACR 2,000 ㎎/㎏ treated mice.
After 18 hrs of fixation, paraffin embedding was conducted and 3-4 μm sections were prepared by routine histological methods. Representative sections of each specified organs were stained with Hematoxylin & eosin for light micro- scopical examination, and any histological findings were re- corded with representative images using a computer based image analysis program (iSolution FL ver 9.1, IMT i-solution Inc., Quebec, Canada).
The absolute organ weight was measured using automatic electronic balance (Precisa Instrument, Zuerich, Switzer- land), and then relative organ weight (% of body weight) was calculated for the following organs of all experimental animals when they were sacrificed.
The animals were distributed into 10 groups, five mice per group after 9 days of acclimatization based on the body weights (Male: 34.47±1.12 g, ranged in 31.0~37.7 g; Female: 26.09±1.56 g, ranged in 23.4~29.3 g), respectively. The high- est dosage used in the present study was selected as 2, 000 ㎎/㎏ in a volume of 20 ml, the limited dosages of rodents and the recommended oral dose volume in mouse[6, 16, 24] using distilled water as vehicle, and 1,000 and 500 ㎎/㎏ were selected as middle and lower dosage groups recom- mended by KFDA Guidelines [16].
In addition, each female and male vehicle controls were added with ACR 2, 000 ㎎/㎏ treated female and male groups. The test article was single orally administered using a zonde attached to 1ml syringe, and only 20 ml/kg of distilled water was once ad- ministered by gastric gavage, instead of test solutions in ve- hicle control mice.
대상 데이터
Measured organs: Lung, Heart, Thymus, Liver, Left Kidney, Left Adrenal Gland, Spleen, Left Testis/Ovary, Splenic lobe of Pancreas, Brain, Left Epididymis/total Uterus and Left Submandibular Lymph node (Total 14 organs).
데이터처리
Variance homogeneity was examined using the Levene test [19]. If the Levene test indicated no significant deviations from variance homogeneity, the obtain data were analyzed by one way ANOVA test followed by Scheffe test to determine which pairs of group comparison were sig- nificantly different. In case of significant deviations from variance homogeneity were observed at Levene test, a non-parametric comparison test, Kruskal-Wallis H test was conducted.
이론/모형
Variance homogeneity was examined using the Levene test [19]. If the Levene test indicated no significant deviations from variance homogeneity, the obtain data were analyzed by one way ANOVA test followed by Scheffe test to determine which pairs of group comparison were sig- nificantly different.
성능/효과
As the results of single oral treatment of fACR, no treat- ment related mortalities were observed within 14 days after end of treatment up to 2,000 ㎎/㎏, the limited dosage of rodents in the both genders, and also no fACR treatment related changes on the body and organ weights, clinical signs, necropsy and histopathological findings were detected in this experiment. In addition, no ACR 2,000 ㎎/㎏ treat- ment related mortalities, clinical signs, body and organ weights, gross and histopathological findings were also ob- served as compared with equal genders of vehicle control, in the present study.
In addition, most of them were also observed in normal mice as sporadic accidental findings [3, 18, 28]. In addition, slight kidney cyst formation was de- tected in 1 female (1/5; 20%) mouse treated with fACR 500 ㎎/㎏ were also difficult to considered as fACR treatment related toxicological signs, because no obvious dos- age-dependencies were detected in this study.
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