본문 바로가기
HOME> 보고서 > 보고서 검색상세

보고서 상세정보

석창포의 90일 반복투여독성 및 유전독성시험
90 days Repeated Dose Toxicity and genotoxicity test of Acorus gramineus

  • 과제명

    천연물 등의 13주 반복투여 및 유전독성시험(석창포, 황련, 단삼, 오수유)

  • 주관연구기관

    바이오톡스텍(주)

  • 연구책임자

    이혜영

  • 보고서유형

    최종보고서

  • 발행국가

    대한민국

  • 언어

    한국어

  • 발행년월

    2009-11

  • 과제시작년도

    2008

  • 주관부처

    식품의약품안전청

  • 사업 관리 기관

    식품의약품안전청
    Korea Food & Drug Administration

  • 등록번호

    TRKO201000015194

  • 과제고유번호

    1475003728

  • 키워드

    석창포.조제물분석.반복독성.유전독성.Acorus gramineus.Validation.repeated dose toxicity.genotoxicity.

  • DB 구축일자

    2013-04-18

  • 초록 


    [1] Validation of analytical method and determination of the stability of Acorus gramineus‘s formulation
    This study was perfor...

    [1] Validation of analytical method and determination of the stability of Acorus gramineus‘s formulation
    This study was performed to validate the analytical method of formulations for Acorus gramineus by high performance liquid chromatograph. The results of systemsuitability, linearity, specification, and intra-day were satisfied with a criterion. And, formulations were homogeneous each phase as upper, middle, and bottom. Formulations that were formulated by high concentration (400 mg/mL) and low concentration (1 mg/mL) were stable for 4 hours at room temperature and for 7 days under refrigeration. In autosampler, the treated samples for analysis were stable for 19 hours in low and high concentrations, respectively.
    [2] 2-week ora l dose range finding study of Acorus gramineus
    This study was performed to determine the dose levels for performing 90 days repeated dose toxicity study in male and female F344 (F344NSlc) rats. The ty stsubstance, Acorus gramineus, was administered orally 12 times to 6-week male and female F344 rats at the doses of 125, 250, 500, 1000, and 2000 mg/kg for 2 weeks. The control group was administered the vehicle (Water for injection) alone. Each group had 5 animals per sex.
    Monitoring clinical signs and measuring body weight were conducted throughout the study. At the end of dosing, all animals were necropsied with major organs being weighed, and histopathological findings were examined. There were no deaths in all animals. No abnormal clinical signs were observed in the male and female treatment groups. No significant difference was observed on the body weights and organ weights in the male and female treatment groups. At necropsy, no treatment-related macroscopic
    findings were noted in all animals. There were no treatment-related histopathological findings in the male and female 2000 mg/kg groups.
    As a result of the 14-day dose range finding study of Acorus gramineus, there were no toxicological signs in the males and females treated with 2000 mg/kg test substance. Therefore, the high dose level for 13 weeks repeated dose toxicity study is considered to be 2000 mg/kg.
    [3] 13-week repeated ora l dose toxicity study of Acorus gramineus
    This study was conducted to assess the toxic reaction of the test substance, Acorus gramineus, when administered five times per week to 6-week-old F344 (F344NSlc) rats by oral gavage for a period of 13 weeks. Test groups consisted of five dosed groups at dose levels of 25, 74, 222, 667 and 2000 mg/kg plus the control. Observations of clinical signs, measurements of body weight and food consumption, urinalysis, and vaginal cytology were conducted throughout the study. At the end of dosing, blood
    samples were collected for hematological and blood chemistry examinations. All animals were necropsied with major organs being weighed; sperm morphology and histopathological findings were examined. Salivation was observed sporadically and temporarily before dosing in several males and
    females of 2000 mg/kg treatment group. No statistically significant differences in the examinations of body weights, food consumption, urinalysis, hematology, blood chemistry, organ weights, vaginal cytology, sperm morphology, necropsy and histopathological findings were noted in males and
    females dosed with the test substance. Based on these results, the no observed adverse effect level (NOAEL) of Acorus gramineus in 13-week oral repeated dose toxicity study to male and female rats was
    considered to be greater than 2000 mg/kg.
    [4] Bacterial reverse mutation test of Acorus gramineus
    This study was designed to examine the mutagenic potential of using Salmonella typhimurium (TA98,TA100,TA1535, and TA1537) and Escherichiacoli (WP2uvrA (pKM101)) strains.
    Test substance was dissolved in Water for injection. In dose range finding test, the growth inhibition was not observed at all dose levels, 5000 μg/plate and lower seven dose levels. Accordingly, bacterial reverse mutation test was performed at the dose levels of 5000, 2500, 1250, 625 and 313 μg/plate.
    As a result of bacterial reverse mutation test, the mean number of revertant colonies was less than twice compared with the negative control values at all dose levels of the test substance in the absence and presence of metabolic activation systems, without a dose-related increase. Neither cell growth inhibition nor deposition of the test substance was observed in all plates in treated groups. Bacterial reverse mutation test was carried out twice to validate the reliability of data. In the positive control group, the number of revertant colonies was distinctly increased as compared with that of the negative control group. In conclusion, the test substance, test substance did not show the mutagenic potential
    under the conditions of this study.
    [5] In vitro chromosome aberration test of Acorus gramineus using mammalian cultured cell
    This study was conducted to examine the potential of the test substance, Acorus gramineus, to induce chromosome aberrations in Chinese hamster lung cell (CHL/IU). As a result of a growth inhibition test, cytotoxicity was not observed for the short time treatment both without and with metabolic activations. but cytotoxicity was observed for and 4342.6 μg/mL in the continuous treatment. Therefore, the highest dose for in vitro chromosome aberration test was determined as e highesmL for the short time treatment both without and with metabolic activations. and the highest dose for was determined as
    44highesmL in the continuous treatment. Those were sequentially diluted by the geometric ratio of 2 to produce 3 additional lower doses accompanied by the negative and positive control groups.
    esult of main test, the structural chromosome aberration cells was more than 5% in the short time treatment with and without and in the continuous treatment of 4400 μg/mL of the test substance. And the frequency of chromosome aberration cells was more than 10% in the short time treatment both without and with metabolic activations and in the continuous treatment of 5000 μg/mL of the test substance, which considered to be positive significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the test substance in the short time treatment without metabolic activation and 4400 μg/mL in the continuous treatment, respectively, and at 5000 μg/mL in the short time treatment
    with metabolic activation, compared with the negative control (Cochran-Armitage trend test, p<0.05).
    As a result of main test, 5000 μg/mL in the short time treatment without and with metabolic activations and 4400 μg/mL in the continuous treatment was juged as positive. Accoding to the result, [1] Validation of analytical method and determination of the stability of Acorus gramineus‘s formulation
    This study was performed to validate the analytical method of formulations for Acorus gramineus by high performance liquid chromatograph. The results of systemsuitability, linearity, specification, and intra-day were satisfied with a criterion. And, formulations were homogeneous each phase as upper, middle, and bottom. Formulations that were formulated by high concentration (400 mg/mL) and low concentration (1 mg/mL) were stable for 4 hours at room temperature and for 7 days under refrigeration. In autosampler, the treated samples for analysis were stable for 19 hours in low and high concentrations, respectively.
    [2] 2-week ora l dose range finding study of Acorus gramineus
    This study was performed to determine the dose levels for performing 90 days repeated dose toxicity study in male and female F344 (F344NSlc) rats. The ty stsubstance, Acorus gramineus, was administered orally 12 times to 6-week male and female F344 rats at the doses of 125, 250, 500, 1000, and 2000 mg/kg for 2 weeks. The control group was administered the vehicle (Water for injection) alone. Each group had 5 animals per sex.
    Monitoring clinical signs and measuring body weight were conducted throughout the study. At the end of dosing, all animals were necropsied with major organs being weighed, and histopathological findings were examined. There were no deaths in all animals. No abnormal clinical signs were observed in the male and female treatment groups. No significant difference was observed on the body weights and organ weights in the male and female treatment groups. At necropsy, no treatment-related macroscopic
    findings were noted in all animals. There were no treatment-related histopathological findings in the male and female 2000 mg/kg groups.
    As a result of the 14-day dose range finding study of Acorus gramineus, there were no toxicological signs in the males and females treated with 2000 mg/kg test substance. Therefore, the high dose level for 13 weeks repeated dose toxicity study is considered to be 2000 mg/kg.
    [3] 13-week repeated ora l dose toxicity study of Acorus gramineus
    This study was conducted to assess the toxic reaction of the test substance, Acorus gramineus, when administered five times per week to 6-week-old F344 (F344NSlc) rats by oral gavage for a period of 13 weeks. Test groups consisted of five dosed groups at dose levels of 25, 74, 222, 667 and 2000 mg/kg plus the control. Observations of clinical signs, measurements of body weight and food consumption, urinalysis, and vaginal cytology were conducted throughout the study. At the end of dosing, blood
    samples were collected for hematological and blood chemistry examinations. All animals were necropsied with major organs being weighed; sperm morphology and histopathological findings were examined. Salivation was observed sporadically and temporarily before dosing in several males and
    females of 2000 mg/kg treatment group. No statistically significant differences in the examinations of body weights, food consumption, urinalysis, hematology, blood chemistry, organ weights, vaginal cytology, sperm morphology, necropsy and histopathological findings were noted in males and
    females dosed with the test substance. Based on these results, the no observed adverse effect level (NOAEL) of Acorus gramineus in 13-week oral repeated dose toxicity study to male and female rats was
    considered to be greater than 2000 mg/kg.
    [4] Bacterial reverse mutation test of Acorus gramineus
    This study was designed to examine the mutagenic potential of using Salmonella typhimurium (TA98,TA100,TA1535, and TA1537) and Escherichiacoli (WP2uvrA (pKM101)) strains.
    Test substance was dissolved in Water for injection. In dose range finding test, the growth inhibition was not observed at all dose levels, 5000 μg/plate and lower seven dose levels. Accordingly, bacterial reverse mutation test was performed at the dose levels of 5000, 2500, 1250, 625 and 313 μg/plate.
    As a result of bacterial reverse mutation test, the mean number of revertant colonies was less than twice compared with the negative control values at all dose levels of the test substance in the absence and presence of metabolic activation systems, without a dose-related increase. Neither cell growth inhibition nor deposition of the test substance was observed in all plates in treated groups. Bacterial reverse mutation test was carried out twice to validate the reliability of data. In the positive control group, the number of revertant colonies was distinctly increased as compared with that of the negative control group. In conclusion, the test substance, test substance did not show the mutagenic potential
    under the conditions of this study.
    [5] In vitro chromosome aberration test of Acorus gramineus using mammalian cultured cell This study was conducted to examine the potential of the test substance, Acorus gramineus, to induce chromosome aberrations in Chinese hamster lung cell (CHL/IU). As a result of a growth inhibition test, cytotoxicity was not observed for the short time treatment both without and with metabolic activations. but cytotoxicity was observed for and 4342.6 μg/mL in the continuous treatment. Therefore, the highest dose for in vitro chromosome aberration test was determined as e highesmL for the short time treatment both without and with metabolic activations. and the highest dose for was determined as
    44highesmL in the continuous treatment. Those were sequentially diluted by the geometric ratio of 2 to produce 3 additional lower doses accompanied by the negative and positive control groups.
    esult of main test, the structural chromosome aberration cells was more than 5% in the short time treatment with and without and in the continuous treatment of 4400 μg/mL of the test substance. And the frequency of chromosome aberration cells was more than 10% in the short time treatment both without and with metabolic activations and in the continuous treatment of 5000 μg/mL of the test substance, which considered to be positive significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the test substance in the short time treatment without metabolic activation and 4400 μg/mL in the continuous treatment, respectively, and at 5000 μg/mL in the short time treatment
    with metabolic activation, compared with the negative control (Cochran-Armitage trend test, p<0.05).
    As a result of main test, 5000 μg/mL in the short time treatment without and with metabolic activations and 4400 μg/mL in the continuous treatment was juged as positive. Accoding to the result, [1] Validation of analytical method and determination of the stability of Acorus gramineus‘s formulation
    This study was performed to validate the analytical method of formulations for Acorus gramineus by high performance liquid chromatograph. The results of systemsuitability, linearity, specification, and intra-day were satisfied with a criterion. And, formulations were homogeneous each phase as upper, middle, and bottom. Formulations that were formulated by high concentration (400 mg/mL) and low concentration (1 mg/mL) were stable for 4 hours at room temperature and for 7 days under refrigeration. In autosampler, the treated samples for analysis were stable for 19 hours in low and high concentrations, respectively.
    [2] 2-week ora l dose range finding study of Acorus gramineus
    This study was performed to determine the dose levels for performing 90 days repeated dose toxicity study in male and female F344 (F344NSlc) rats. The ty stsubstance, Acorus gramineus, was administered orally 12 times to 6-week male and female F344 rats at the doses of 125, 250, 500, 1000, and 2000 mg/kg for 2 weeks. The control group was administered the vehicle (Water for injection) alone. Each group had 5 animals per sex.
    Monitoring clinical signs and measuring body weight were conducted throughout the study. At the end of dosing, all animals were necropsied with major organs being weighed, and histopathological findings were examined. There were no deaths in all animals. No abnormal clinical signs were observed in the male and female treatment groups. No significant difference was observed on the body weights and organ weights in the male and female treatment groups. At necropsy, no treatment-related macroscopic
    findings were noted in all animals. There were no treatment-related histopathological findings in the male and female 2000 mg/kg groups.
    As a result of the 14-day dose range finding study of Acorus gramineus, there were no toxicological signs in the males and females treated with 2000 mg/kg test substance. Therefore, the high dose level for 13 weeks repeated dose toxicity study is considered to be 2000 mg/kg.
    [3] 13-week repeated ora l dose toxicity study of Acorus gramineus
    This study was conducted to assess the toxic reaction of the test substance, Acorus gramineus, when administered five times per week to 6-week-old F344 (F344NSlc) rats by oral gavage for a period of 13 weeks. Test groups consisted of five dosed groups at dose levels of 25, 74, 222, 667 and 2000 mg/kg plus the control. Observations of clinical signs, measurements of body weight and food consumption, urinalysis, and vaginal cytology were conducted throughout the study. At the end of dosing, blood
    samples were collected for hematological and blood chemistry examinations. All animals were necropsied with major organs being weighed; sperm morphology and histopathological findings were examined. Salivation was observed sporadically and temporarily before dosing in several males and
    females of 2000 mg/kg treatment group. No statistically significant differences in the examinations of body weights, food consumption, urinalysis, hematology, blood chemistry, organ weights, vaginal cytology, sperm morphology, necropsy and histopathological findings were noted in males and
    females dosed with the test substance. Based on these results, the no observed adverse effect level (NOAEL) of Acorus gramineus in 13-week oral repeated dose toxicity study to male and female rats was
    considered to be greater than 2000 mg/kg.
    [4] Bacterial reverse mutation test of Acorus gramineus
    This study was designed to examine the mutagenic potential of using Salmonella typhimurium (TA98,TA100,TA1535, and TA1537) and Escherichiacoli (WP2uvrA (pKM101)) strains.
    Test substance was dissolved in Water for injection. In dose range finding test, the growth inhibition was not observed at all dose levels, 5000 μg/plate and lower seven dose levels. Accordingly, bacterial reverse mutation test was performed at the dose levels of 5000, 2500, 1250, 625 and 313 μg/plate.
    As a result of bacterial reverse mutation test, the mean number of revertant colonies was less than twice compared with the negative control values at all dose levels of the test substance in the absence and presence of metabolic activation systems, without a dose-related increase. Neither cell growth inhibition nor deposition of the test substance was observed in all plates in treated groups. Bacterial reverse mutation test was carried out twice to validate the reliability of data. In the positive control group, the number of revertant colonies was distinctly increased as compared with that of the negative control group. In conclusion, the test substance, test substance did not show the mutagenic potential
    under the conditions of this study.
    [5] In vitro chromosome aberration test of Acorus gramineus using mammalian cultured cell This study was conducted to examine the potential of the test substance, Acorus gramineus, to induce chromosome aberrations in Chinese hamster lung cell (CHL/IU). As a result of a growth inhibition test, cytotoxicity was not observed for the short time treatment both without and with metabolic activations. but cytotoxicity was observed for and 4342.6 μg/mL in the continuous treatment. Therefore, the highest dose for in vitro chromosome aberration test was determined as e highesmL for the short time treatment both without and with metabolic activations. and the highest dose for was determined as
    44highesmL in the continuous treatment. Those were sequentially diluted by the geometric ratio of 2 to produce 3 additional lower doses accompanied by the negative and positive control groups.
    esult of main test, the structural chromosome aberration cells was more than 5% in the short time treatment with and without and in the continuous treatment of 4400 μg/mL of the test substance. And the frequency of chromosome aberration cells was more than 10% in the short time treatment both without and with metabolic activations and in the continuous treatment of 5000 μg/mL of the test substance, which considered to be positive significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the significant dose-dependent increases in the incidence were observed at 5000 μg/mL of the test substance in the short time treatment without metabolic activation and 4400 μg/mL in the continuous treatment, respectively, and at 5000 μg/mL in the short time treatment
    with metabolic activation, compared with the negative control (Cochran-Armitage trend test, p<0.05).
    As a result of main test, 5000 μg/mL in the short time treatment without and with metabolic activations and 4400 μg/mL in the continuous treatment was juged as positive. Accoding to the result, $D_{20}$ and TR were calculated that were of 8.9 and 5.0 in the short time treatment without, 5.2 and 5.0 in the short time treatment with, and 3.4 and 7.1 in the continuous treatment.
    The incidences of chromosome aberrations were 10% and above in the positive control groups. Therefore, this study was considered to be valid and evaluation of the study results was appropriate.
    Base upon these results, the test substance, Acorus gramineus, was judged to be positive according to the criteria by Sofuni et al. under the conditions of this study. In the positive control group, the structural chromosome aberration was significantly increased (Fisher’s exact test, p<0.05). The test substance, Acorus gramineus, shown the chromosome aberrations in the short time treatment without and with metabolic activation and the continuous treatment using Chinese hamster lung cell (CHL/IU) under the conditions of this study.
    [6] In vivo micronucleus test of Acorus gramineus in mice
    This study was performed to determine the mutagenic potential of the test substance, Auorus gramineus , in the micronucleus test using bone marrow cells when administered once by the oral gavage to male ICR mice. For the determining of the highest dose and bone marrow collection time, the dose range finding test and bone marrow collection time determining test were performed. As a
    result, the highest dose for in vivo micro nucleus test was determined as 2000 mg/kg and the bone marrows were collected at 24 hours after the administration. In vivo micronucleus test was conducted at 3 dose levels of 500, 1000 and 2000 mg/kg of the test substance. In addition, the negative and positive control groups were included in the test. Five mice per group were used. As a result of main test, the animal death and clinical signs were not observed at all doses of test substance.
    There was not body weight change during the test period. There were no statistically significant increase and dose-dependency (p<0.05) in appearance ratio of micronucleated polychromatic erythrocytes (MNPCE) in polychromatic erythrocytes (PCE) for all test substance groups as compared with that of the negative control group. The proportions of PCE in total erythrocyte were also no significant difference between test substance group and negative control group. In the positive control, the appearance ratio of MNPCE in PCE was significantly increased as compared with that of the negative control group. Based on these results, it was concluded that the test substance, Acorus gramineus, did not influence on the micronucleus formation in the bone marrow cells of mice under the
    condition of this study. and TR were calculated that were of 8.9 and 5.0 in the short time treatment without, 5.2 and 5.0 in the short time treatment with, and 3.4 and 7.1 in the continuous treatment.
    The incidences of chromosome aberrations were 10% and above in the positive control groups. Therefore, this study was considered to be valid and evaluation of the study results was appropriate.
    Base upon these results, the test substance, Acorus gramineus, was judged to be positive according to the criteria by Sofuni et al. under the conditions of this study. In the positive control group, the structural chromosome aberration was significantly increased (Fisher’s exact test, p<0.05). The test substance, Acorus gramineus, shown the chromosome aberrations in the short time treatment without and with metabolic activation and the continuous treatment using Chinese hamster lung cell (CHL/IU) under the conditions of this study.


    [1] 석창포의 조제물의 농도 분석법 Validation 및 안정성 확인시험
    본 연구에서는 석창포의 조제물 분석법 검증을 위해 HPLC를 이용하여 분석법 validation을 수행하였다. 시스템 적합성, 직선성, 특이성, 일...

    [1] 석창포의 조제물의 농도 분석법 Validation 및 안정성 확인시험
    본 연구에서는 석창포의 조제물 분석법 검증을 위해 HPLC를 이용하여 분석법 validation을 수행하였다. 시스템 적합성, 직선성, 특이성, 일내재현성 측정결과 판정기준을 모두 만족하였으며, 조제물은 상, 중, 하층에서 모두 균질하였다. 고농도 400 mg/mL 와 저농도 1 mg/mL 농도의 조제물은 실온에서 4시간, 냉장에서 7일간 안정한 것으로 확인되었으며, 측정을 위해 처리된 시료는 autosampler 내에서 19 시간 동안 안정하였다.
    [2] 석창포의 랫드를 이용한 2주 반복 경구투여 용량결정시험
    석창포의 90일 반복투여 독성시험시 용량설정 근거로 사용하고자, 6주령 (투여개시시 주령) 암수 F344 (F344NSlc) 랫드에 125, 250, 500, 1000, 2000 mg/kg의 5개의 용량으로 14일간 총 12회 강제 경구 투여하였다. 대조군에는 주사용수를 투여하였다. 일반증상 관찰, 체중측정, 부검시 장기중량 측정과 육안적 검사를 실시하였고, 주요장기에 대해 조직병리학적 검사를 수행하였다. 암수 시험물질투여군에서 일반증상, 체중, 장기중량 및 부검에서 시험물질에 의한 영향은 관찰 되지 않았다. 조직병리학적 검사 결과, 암수 2000 mg/kg 투여군에서 시험물질에 의한 소견은 관찰되지 않았다. 이상으로 석창포를 2주 경구 반복 투여한 결과, 암수 2000 mg/kg 투여군에서 시험물질 투여에 기인한 독성변화가 관찰되지 않았다. 따라서 13주 반복투여 독성시험의 고용량은 2000mg/kg으로 설정해야 할 것으로 판단된다.
    [3] 석창포의 랫드를 이용한 13주 반복 경구투여 독성시험
    석창포의 13주 반복투여 독성시험에 대한 안전성을 평가하고자, F344 (F344NSlc)계 암수 랫드에 시험물질을 25, 74, 222, 667 및 2000 mg/kg의 5개의 용량으로 13주간 주 5회 경구 투여하였다. 대조군에는 주사용수를 투여하였다.
    관찰기간동안 일반증상관찰, 체중측정, 사료섭취량 측정, 뇨검사, 암컷의 성주기 검사를 실시하고, 관찰기간 종료 후 혈액 및 혈액생화학적 검사, 수컷의 부고환 미부내 정자검사, 장기의 중량측정, 부검시 육안적 검사 및 조직병리학적 검사를 수행하였다. 암수 2000 mg/kg 투여군의 일부개체에서 투여전 유연이 일시적 또는 산발적으로 관찰되었다. 체중, 사료섭취량, 뇨검사, 혈액학적 검사, 혈액생화학적 검사, 암컷의 성주기 검사, 수컷의 부고환 미부내 정자검사, 장기중량, 부검 및 조직병리학적 검사에 있어서, 암수 시험물질투여군에서 시험물질 투여에 기인한 독성변화는 관찰되지 않았다. 이상으로 석창포에 대한 암수 랫드를 이용한 13주 경구 반복투여 독성시험을 실시한 결과, 암수 시험물질투여군에서 독성변화가 인정되지 않았다. 따라서 본 시험 조건 하에서의 무독성량 (NOAEL)은 암수 모두 2000 mg/kg을 상회하는 것으로 판단된다.
    [3] 석창포의 세균을 이용한 복귀돌연변이시험
    시험물질인 석창포의 유전자돌연변이 유발성을 히스티딘 요구성인 살모넬라균 (TA98, TA100, TA1535, TA1537) 및 트립토판 요구성인 대장균 (WP2uvrA(pKM101))을 이용하여 대사활성화비 존재하 및 존재하인 각각의 경우에 대해 검토하였다. 용량설정시험의 결과, 대사활성화비존재하 및 존재하의 모든 균주에서 생육저해가 관찰되지 않았다.
    시험물질의 침전은 대사활성화의 유무에 관계없이 이용된 모든 균주의 모든 용량에서 관찰되지 않았다.
    본시험의 최고용량은 대사활성화비존재하 및 존재하의 5000 μg/plate를 최고용량으로 설정하고 최고용량을 포함한 5용량(공비2)을 시험물질군으로 설정하였다. 또한 음성대조군 및 양성대조군을 설정하였다.
    본시험의 결과, 각 균주에 있어 시험물질군의 복귀변이콜로니수는 대사활성화 비존재하 및 존재 하에서 용량의존적으로 증가되지 않았으며 음성대조군과 비교하여 2배 이상의 증가를 보이지 않았다. 또한 대사활성화비존재하 및 존재하에서 모든 균주의 모든 용량에서 시험물질에 의한 생육저해 및 침전은 관찰되지 않았다.
    한편, 각 균주에서 양성대조군의 복귀변이콜로니수는 음성대조군과 비교하여 확실한 증가가 인정되었다. 이상의 결과로부터, 본 시험조건에서 시험물질인 석창포의 유전자돌연변이 유발성은 음성으로 판정 되었다.
    [4] 석창포의 포유류 배양세포를 이용한 염색체이상시험
    시험물질인 석창포의 염색체이상 유발성 유무를 검토하기 위하여 Chinese Hamster Lung (CHL/IU) 배양세포를 이용한 염색체이상시험을 실시하였다. 세포증식억제시험의 결과, 대사활성화비존재하의 24시간 처리의 경우에서 세포독성을 나타내었기 때문에 $IC_5_0$를 구한 결과, 4342.6 μg/mL이었다. 따라서, 단시간처리법으로서 대사활성화비존재하 및 대사활성화존재하의 경우는 5000 μg/mL, 연속처리법으로서 대사활성화비존재하의 24시간 처리의 경우는 4400 μg/mL를 본시험의 최고용량으로 설정하고 이하 공비 2로 총 3용량의 시험물 질군을 설정하였다. 또한 음성대조군 및 양성대조군을 설정하였다. 본시험의 결과, 단시간처리법으로서 대사활성화비존재하 및 존재하의 5000 μg/mL과 연속처리법 대사활성화비존재하의 4400 μg/mL에서 구조이상을 가진 세포의 출현 빈도가 10% 이상으로 염색체이상 유발작용이 인정되었다. 그러나 수적이상을 가진 세포의 출현빈도는 5% 미만으로 염색체이상 유발작용이 확인되지 않았다.
    또한, 구조이상을 가지는 세포의 출현빈도는 용량의존적으로 증가되었으며, 음성 대조군과 시험 물질군간 비교시 단시간처리법의 대사활성화비존재하의 경우 및 존재하의 경우에는 5000 μg/mL 과 연속처리법으로서 대사활성화 비존재하의 24시간처리의 경우 4400 μg/mL에서 통계학적으로 유의한 증가가 확인되었다 (Cochran-Armitage trend test, p<0.05).
    한편, 양성대조군의 구조이상을 가진 세포의 출현빈도는 현저하게 증가하였다. 이상의 결과로부터 본 시험조건하에서 시험물질인 석창포는 단시간처리법의 대사 활성화비존재 하 및 존재하, 연속처리법의 대사활성화비존재하 모두에서 Chinese Hamster Lung (CHL/IU) 배양 세포에 대해 염색체이상을 유발하는 것으로 판단된다
    [6] 석창포의 마우스를 이용한 소핵시험
    시험물질인 석창포의 마우스 골수세포에 대한 소핵유발 유무를 평가하기 위하여 시험을 실시하였다.
    최고용량 및 검체제작시간을 설정하기 위하여, 용량설정시험 및 검체제작시간 설정시험을 실시하였다. 이러한 결과에 따라 본시험에서의 최고용량은 2000 mg/kg으로 투여 후 24시간을 골수세포 채취시간으로 결정하였다.
    본시험은 시험물질 500, 1000 및 2000 mg/kg의 총 3용량을 설정하였으며 각 군당 5마리의 마우스를 사용하였다. 또한 음성대조군 및 양성대조군을 두었다. 본시험의 결과, 모든 시험물질군에서 시험물질 투여에 의한 사망동물은 없었고 독성증상도 관찰 되지 않았다.
    체중변화는 실험 전 기간을 통하여 유의성 있는 변화는 관찰되지 않았다. 시험물질군에서 다염성적혈구(Polychromatic erythrocyte, PCE)중 소핵다염성적혈구(Micronucleated polychromatic erythrocyte, MNPCE)의 빈도는 모든 용량에서 음성대조군과 비교하여 통계학적으로 유의한 증가(p<0.05)가 관찰되지 않았다. 또한 모든 시험물질군에서 총 적
    혈구 중 다염성적혈구의 비율은 음성대조군과 비교하여 유의한 차이는 관찰되지 않았다. 한편, 양성대조군의 다염성적혈구 중 소핵다염성적혈구의 출현빈도는 음성대조군과 비교하여 현저한 증가가 인정되었다.
    이상의 결과로 석창포는 본 시험의 조건하에서 마우스 골수세포의 소핵유발에 영향을 주지 않는 것으로 판단된다.


  • 목차(Contents) 

    1. 표지 ...1
    2. 최종보고서 ...2
    3. 제출문 ...3
    4. 목차 ...4
    5. I. 연구개발결과 요약문 ...5
    6. 국문요약문 ...5
    7. SUMMARY ...10
    8. II. 총괄연구개발과제 연구결과 ...16
    9. 제1장 총괄연구개발과제의 최종 연구개발 목표 ...1...
    1. 표지 ...1
    2. 최종보고서 ...2
    3. 제출문 ...3
    4. 목차 ...4
    5. I. 연구개발결과 요약문 ...5
    6. 국문요약문 ...5
    7. SUMMARY ...10
    8. II. 총괄연구개발과제 연구결과 ...16
    9. 제1장 총괄연구개발과제의 최종 연구개발 목표 ...16
    10. 1절. 총괄연구개발과제의 목표...16
    11. 2절. 총괄연구개발과제의 목표달성도...17
    12. 3절. 국내.외 기술개발 현황...18
    13. 제2장 총괄연구개발과제의 최종 연구개발 내용 및 방법 ...19
    14. 1절. 연구내용...19
    15. 제3장 총괄연구개발과제의 최종 연구개발 결과 ...59
    16. 1절. 석창포의 조제물의 농도 분석법 Validation 및 안정성 확인시험...59
    17. 2절. 석창포의 2주 반복 경구투여 용량결정시험...75
    18. 3절. 석창포의 13주 반복 경구투여 독성시험...96
    19. 4절. 석창포의 세균을 이용한 복귀돌연변이시험...202
    20. 5절. 석창포의 포유류 배양세포를 이용한 염색체이상시험...209
    21. 6절. 석창포의 마우스를 이용한 소핵시험...212
    22. 제4장 총괄연구개발과제의 연구결과 고찰 및 결론 ...217
    23. 가. 석창포의 조제물의 농도 분석법 Validation 및 안정성 확인시험...217
    24. 나. 석창포의 2주 반복 경구 투여 용량결정시험...217
    25. 다. 석창포의 13주 반복 경구 투여 독성시험 ...217
    26. 라. 석창포의 세균을 이용한 복귀돌연변이시험...218
    27. 마. 석창포의 포유류 배양세포를 이용한 염색체이상시험...218
    28. 바. 석창포의 마우스를 이용한 소핵시험...218
    29. 제5장 총괄연구개발결과의 연구성과 ...219
    30. 5.1. 활용성과...219
    31. 5.2 활용계획...220
    32. 제6장 기타중요변경사항 ...220
    33. 제7장 참고문헌 ...220
    34. 제8장 첨부서류 ...221
  • 참고문헌

    1. 전체(0)
    2. 논문(0)
    3. 특허(0)
    4. 보고서(0)

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역