赤苷脉通注射液的致突变研究

目的探讨中药制剂赤苷脉通注射液的致突变性。方法采用鼠伤寒沙门氏组氨酸营养缺陷型菌株回复突变实验(Ames实验)、中国仓鼠肺成纤维细胞(CHL)染色体畸变实验和小鼠骨髓微核实验来检测赤苷脉通注射液的致突变作用。结果 Ames实验中,赤苷脉通注射液在312.5～5 000μg.皿-1剂量范围内,无论加或不加S9,鼠伤寒沙门氏菌组氨酸缺陷型TA97,TA98,TA100,TA102和TA1535 5株菌的回复突变菌落数均未出现剂量依赖性的增加;染色体畸变实验中,非活化条件或代谢活化条件下,药物质量浓度为1 200,600和300μg.mL-1时,细胞的染色体畸变率均未出现剂量依赖性增加;微核实验中,在1 150,575和287.5mg.kg-1剂量组中均未见骨髓中含微核的嗜多染红细胞数增加。结论在该实验室条件下,Ames实验、CHL细胞染色体畸变实验和小鼠骨髓微核实验结果均为阴性,即中药制剂赤苷脉通注射液无潜在的遗传毒性。     

中药浮海石致突变的实验研究

目的研究中药浮海石致突变性。方法本试验采用鼠伤寒沙门菌回复突变试验(Ames试验),哺乳动物培养细胞(CHL)染色体畸变试验和小鼠骨髓细胞微核试验观察了浮海石的致突变作用。结果浮海石生理盐水浸提液0.5~5000μg/皿剂量下Ames试验结果阴性;250~1000μg/mL剂量下对CHL细胞无损伤作用,10~40g/kg剂量下对小鼠骨髓细胞染色体无致畸变作用。结论浮海石无致突变作用,用于临床是安全的。     

Evaluation of the mutagenic and genotoxic potential of ubiquinol

Ubiquinol (the reduced form of coenzyme Q(10)) is the two-electron reduction product of ubiquinone (the oxidized form of coenzyme Q(10)), and has been shown to be an integral part of living cells, where it functions as an antioxidant in both mitochondria and lipid membranes. To provide information to enable a Generally Regarded as Safe (GRAS) evaluation for the use of ubiquinol in selected foods, a series of Organisation of Economic Cooperation and Development (OECD) and good laboratory practice (GLP) toxicological studies was conducted to evaluate the mutagenic and genotoxic potential of Kaneka QH brand of ubiquinol. Ubiquinol did not induce reverse mutations in Salmonella typhimurium strains TA100, TA1535, TA98, and TA1537 and Escherichia coli WP2uvrA at concentrations up to 5000 mu g/plate, in either the absence and presence of exogenous metabolic activation by rat liver S9. Likewise, ubiquinol did not induce chromosome aberrations in Chinese hamster lung fibroblast (CHL/IU) cells in short-term (6-h) tests with or without rat liver S9 at concentrations up to 5000 mu g/ml or in a continuous (24-h) treatment test at concentrations up to 1201 mu g/ml. Finally, no mortalities, no abnormal clinical signs, and no significant increase in chromosome damage were observed in an in vivo micronucleus test when administered orally at doses up to 2000 mg/kg/day. Thus, ubiquinol was evaluated as negative in the bacterial reverse mutation, chromosomal aberration, and rat bone marrow micronucleus tests under the conditions of these assays.     

黄蜀葵花提取物金丝桃苷的急性毒性和遗传毒性评价(英文)

研究黄蜀葵花提取物金丝桃苷的急性毒性和遗传毒性,对其安全性进行评价。急性毒性试验中,选用健康BALB/c小鼠40只,雌雄各半,灌胃给药(5000mg/kg),连续观察14天,记录中毒和死亡情况,测定小鼠的半数致死量(LD50)。用目前新药遗传毒性评价中推荐使用的3种试验方法,营养缺陷型鼠伤寒沙门氏菌回复突变试验(Ames试验),中国仓鼠肺成纤维细胞(CHL)染色体畸变试验和小鼠骨髓微核试验研究金丝桃苷的遗传毒性。在急性毒性试验中,所有实验动物都存活,且行为活泼,未见明显异常。Ames试验中,金丝桃苷在加或不加肝微粒体酶(S9)时均未见引起TA97、TA98、TAl00和TAl02试验菌株基因突变(P>0.05)。体外CHL细胞染色体畸变试验中,金丝桃苷在加或不加S9时均未引起CHL细胞的染色体畸变(P>0.05)。小鼠微核试验中,金丝桃苷各剂量组小鼠骨髓多染红细胞微核率与阴性对照组相比,差异无统计学意义(P>0.05)。在本实验条件下,金丝桃苷对于BALB/c小鼠的LD50大于5000mg/kg,金丝桃苷没有遗传毒性。     

Genotoxicity studies of cefmatilen hydrochloride hydrate (S-1090)

Cefmatilen hydrochloride hydrate (S-1090), a new non-ester type of orally active cephem antibiotic synthesized in Shionogi Research Laboratories, was evaluated for its genotoxic potential using three assay systems. In a reverse mutation test with bacteria of Salmonella typhimurium TA100, TA1535, TA98, and TA1537, and Escherichia coli WP2uvrA using the preincubation method, the number of revertant colonies in the S-1090 treated plates was almost equal to that in the negative control plates in all strains with and without metabolic activation system with S9 mix (maximum dose, 100 micrograms/plate in TA98). In a chromosomal aberration test with cultured Chinese hamster lung cells (CHL/IU), S-1090 did not induce structural chromosome aberrations or polyploid cells either in the absence or presence of S9 mix up to the 50% growth inhibition doses. The potential of inducing clastogenicity and/or disruption of mitotic apparatus in vivo by S-1090 was evaluated by a micronucleus test with bone marrow cells of male Jc1:ICR mice. S-1090 suspended in 0.5% aqueous methylcellulose was administered by oral gavage up to 2000 mg/kg/day in single and double dosing groups. No induction of micronucleated polychromatic erythrocytes was observed 24 hr after the last dosing in each group. As all three genotoxicity tests showed negative responses, S-1090 is thought to have no genotoxic potential.     

白障明的诱变性试验研究

作者用鼠伤寒沙门氏菌回变试验,小鼠骨髓红细胞微核试验及CHL细胞染色体畸变试验,对白障明进行了诱变性试验研究。结果表明鼠伤寒沙门氏菌回变试验为阴性,该药可能不诱发基因突变,NIH小鼠骨髓红细胞微核试验及CHL细胞染色体畸变试验均为阴性,说明该药可能不诱发体内,体外染色体损伤作用。     

聚乙二醇修饰降纤酶的遗传毒性评价

目的检测聚乙二醇修饰降纤酶的遗传毒性。方法应用鼠伤寒沙门菌回复突变试验(Ames试验)、体外培养CHO细胞染色体畸变试验和小鼠骨髓微核试验检测聚乙二醇修饰降纤酶的遗传毒性。结果 Ames试验结果显示每平皿100、20、4、0.8、0.16 U各个剂量组,在加或不加S9代谢活化系统时对组氨酸缺陷型鼠伤寒沙门菌TA97、TA98、TA100、TA102及TA1535所诱发的回复突变菌落数均与溶剂对照的突变菌落数相近。体外培养CHO细胞染色体畸变试验结果显示2.5、5.0和10.0 U.mL-1各个剂量组在加S9代谢活化系统于24 h和不加S9代谢活化系统于24 h、48 h培养的CHO细胞染色体畸变率与溶剂对照组比较均无显著差异(P>0.05)。小鼠骨髓微核试验显示425、850、1700 U.kg-1各个剂量组对ICR小鼠的微核诱发率与溶剂对照组比较均无显著差异(P>0.05)。结论聚乙二醇修饰降纤酶对鼠伤寒沙门菌无致突变性,对哺乳动物培养细胞的染色体无致畸变作用,对ICR小鼠无诱发骨髓嗜多染红细胞微核的效应。表明聚乙二醇修饰降纤酶在本实验条件下无遗传毒性。     

崖柏精油致突变作用研究

摘 要 目的：采用鼠伤寒沙门菌回复突变（AMES）试验和哺乳动物微核试验对崖柏精油的遗传毒性进行评价。方法: AMES试验采用TA97、TA98、TA100和TA102四种菌株,对崖柏精油的致突变性进行评价。采用小鼠骨髓噬多染红细胞微核试验,对本品的染色体毒性进行评价。结果: 崖柏精油不同剂量组对TA97、TA98、TA100、TA102四种试验菌株,在加大鼠肝微粒体酶S9和不加S9的情况下,回复突变数均不超过溶剂对照组的1倍,差异无统计学意义（P<0.05）,为诱变阴性。微核试验表明,崖柏精油各剂量组对小鼠骨髓微核率无显著影响（P>0.05）。结论:崖柏精油无明显遗传毒性。     

蚓激酶的诱变实验研究

用鼠伤寒沙门氏菌回复突变试验，小鼠骨髓红细胞微核试验及ＣＨＬ细胞染色体畸变试验，对蚓激酶进行了诱变性试验研究。结果表明鼠伤寒沙门氏菌回复突变试验为阴性，该药可能不诱发基因突变；小鼠骨髓红细胞微核试验及ＣＨＬ细胞染色体畸变试验均为阴性，说明该药可能不诱发体内、体外遗传物质损伤作用。     

国产环丙沙星的诱变性和致癌性研究

应用小鼠骨髓细胞微核试验、CHL细胞染色体畸变试验及叙利亚地鼠胚胎细胞转化试验,对国产环丙沙星进行了诱变性和致癌研究。结果: 环丙沙星在小鼠骨髓细胞微核试验和CHL细胞染色体畸变试验中均为阴性;在细胞转化试验中不诱发SHE细胞发生形态转化。实验结果提示环丙沙星在体内、外不引起染色体损伤;无体外致癌作用。     

Developmental toxicity and genotoxicity studies of wogonin

We studied the developmental toxicities and genotoxic potency of a widely bioactive plant medicine-wogonin in vivo and in vitro. In the in vivo developmental experiments, high dose of wogonin (40mg/kg, intravenous injection) significantly induced the maternal weight gains and affected fetus including bodyweight, resorptions, live birth index and fetal skeletal alterations. In Ames test, no concentration-dependently increased TA98, TA100, and TA102 revertants were detected in wogonin groups whether in presence of metabolic activating enzymes or not. In the chromosome aberration test, wogonin dose-dependently increased structural chromosomal aberrations in CHL cells both with and without S9, even the effect was all judged (-). In micronucleus assay, no significant changes of MNPCE/PCE and PCE/NCE were found on mouse bone marrow micronucleus in wogonin groups. We concluded that wogonin induced developmental toxicities on pregnant mice and fetus, and the genotoxicities were positive. However no significant malformation was observed and only in vitro potency of chromosome aberration was weak, which suggested us wogonin could be a relatively safe drug in clinic.     

Genotoxicity testing of sodium formononetin-3′-sulphonate (Sul-F) by assessing bacterial reverse mutation,chromosomal aberrations and micronucleus tests

As part of a safety evaluation, we evaluated the potential genotoxicity of sodium formononetin-3′-sulphonate (Sul-F) using bacterial reverse mutation assay, chromosomal aberrations detection, and mouse micronucleus test. In bacterial reverse mutation assay using five strains of Salmonella typhimurium (TA97, TA98, TA100, TA102 and TA1535), Sul-F (250, 500, 1000, 2000, 4000 μg/plate) did not increase the number of revertant colonies in any tester strain with or without S9 mix. In a chromosomal assay using Chinese hamster lung fibroblast (CHL) cells, there were no increases in either kind of aberration at any dose of Sul-F (400, 800, and 1600 μg/mL) treatment groups with or without S9 metabolic activation. In an in vivo bone marrow micronucleus test in ICR mice, Sul-F at up to 2000 mg/kg (intravenous injection) showed no significant increases in the incidence of micronucleated polychromatic erythrocytes, and the proportion of immature erythrocytes to total erythrocytes. The results demonstrated that Sul-F does not show mutagenic or genotoxic potential under these test conditions.     

In vitro and in vivo genotoxic effects of somatic cell nuclear transfer cloned cattle meat

Although the nutritional composition and health status after consumption of the meat and milk derived from both conventionally bred (normal) and somatic cell nuclear transferred (cloned) animals and their progeny are not different, little is known about their food safeties like genetic toxicity. This study is performed to examine both in vitro (bacterial mutation and chromosome aberration) and in vivo (micronucleus) genotoxicity studies of cloned cattle meat. The concentrations of both normal and cloned cattle meat extracts (0-10×) were tested to five strains of bacteria (Salmonella typhimurium: TA98, TA100, TA1535, and TA1537; Escherichia coli: WP2uvrA) for bacterial mutation and to Chinese hamster lung (CHL/IU) cells for chromosome aberration, respectively. For micronucleus test, ICR mice were divided into five dietary groups: commercial pellets (control), pellets containing 5% (N-5) and 10% (N-10) normal cattle meat, and pellets containing 5% (C-5) and 10% (C-10) cloned cattle meat. No test substance-related genotoxicity was noted in the five bacterial strains, CHL/IU cells, or mouse bone marrow cells, suggesting that the cloned cattle meat potentially may be safe in terms of mutagenic hazards. Thus, it can be postulated that the cloned cattle meat do not induce any harmful genotoxic effects in vitro and in vivo.     

Genotoxicity studies of 2,6-dinitrotoluene (2,6-DNT)

The potential genotoxicity of the rodent liver carcinogen 2,6-dinitrotoluene (2,6-DNT) was evaluated in compliance with the guidelines for genotoxicity studies of drugs (Notification No. 1604, Nov. 1, 1999, Ministry of Health and Welfare, Japan) and the OECD guidelines for the testing of chemicals by performing the bacterial reverse mutation (Ames) assay, the in vitro chromosomal aberration assay, and the in vivo comet assay (alkaline single cell gel electrophoresis) in rat liver. In the Ames assay, 2,6-DNT was moderately positive irrespective of metabolic activation. In the in vitro chromosomal aberration assay, under conditions where the test substance would precipitate out, weak structural aberrations were observed with or without S9 mix at each dose at which the cell growth rate was about 40 to 50%. The in vivo comet assay yielded positive results in rat liver; that is to say, the increases in % tail DNA in liver in the 25 and 50 mg/kg groups were observed statistically significantly and dose-dependent. Our findings are in accordance with previous findings in the in vivo/in vitro unscheduled DNA synthesis (UDS) assay in rat liver and in a young rat liver micronucleus assay, although the rat bone marrow micronucleus assay gave negative results. These results suggest that test systems using liver are a useful method for the in vivo genotoxicity assessment of chemicals that require metabolic activation.     

Mutagenicity of recombinant antihemophilic factor (GC-gamma AHF)

This study was carried out to evaluate the mutagenic potential of recombinant antihemophilic factor VIII (GC-gamma AHF). Salmonella typhimurium (S. typhimurium) reversion assay with/without histidine moiety, chromosomal aberration assay on Chinese hamster lung (CHL) fibroblast cells and in vivo micronucleus assay using mouse bone marrow cells and supravital micronucleus assay using peripheral blood were performed. GC-gamma AHF containing histidine did show inconsistent and irregular mutagenic effects on S. typhimurium TA98, TA100, TA1535 and TA1537 both in the absence and presence of the metabolic activation system, however, GC-gamma AHF without histidine showed no mutagenic effects regardless of the metabolic activation system, thus suggesting that the histidine moiety in GC-gamma AHF might cause inconsistent mutagenic effect. Also GC-gamma AHF did not increase the number of cells having structural or numerical chromosome aberration in the cytogenetic test. In classical and supravital micronucleus assay, no significant increases were observed in the occurrence of micronucleated polychromatic erythrocytes and micronucleated peripheral lymphocytes in male ICR mice. These results strongly indicate that GC-gamma AHF has no genetic toxicity under these experimental conditions.     

Genotoxicity studies OF p-dimethylaminoazobenzene (DAB)

The potential genotoxicity of the rodent liver carcinogen p-dimethylaminoazobenzene (DAB) was evaluated in compliance with the guidelines for genotoxicity studies of drugs (Notification No. 1604, Nov. 1, 1999, Ministry of Health and Welfare, Japan) and the OECD guidelines for testing chemicals. DAB was clearly positive in both the bacterial reverse mutation test (Ames test) and in vitro chromosomal aberration test in the presence of metabolic activation, whereas it was weakly positive at toxic doses in the rat bone marrow micronucleus test. It has been reported that DAB was clearly positive in in vivo genotoxicity tests, i.e., a mouse alkaline single cell gel electrophoresis (comet) assay and a young rat liver micronucleus test. These results suggest that the test system using the liver is effective for in vivo genotoxicity assessment of chemicals that show mutagenicity in in vitro genotoxicity tests in the presence of metabolic activation.     

依立雄胺体内外致突变实验

AIM: To evaluate the genetic effects of epristeride (Epr), a new prospective drug for treating benign prostatic hyperplasia. METHODS: 1) Assaying reverse mutation in histidine nutritional deficiency strain of Salmonella typhimurium 2) detecting chromosome aberrations in Chinese hamster lung cells (CHL); 3) micronucleus assays of mouse bone marrow; 4) counting sperm shape abnormalities 35 d after first ig Epr. RESULTS: 1) The reverse mutation happened at almost the same rate of the negative control. Epr did not induce bacterial mutation. 2) In vitro, the rates of aberration were all below 3%, thus Epr did not induce chromosome damage in CHL. 3) Micronucleated polychromatic erythroblasts (PCE) were not apparently more than those of sovent control, Epr did not induce the formation of micronuclei in PCE. 4) With Epr 818, 682, and 341 mg.kg-1, the head abnormalities of sperms were 5.3% +/- 2.7%, 5.3% +/- 1.9%, and 5.2% +/- 1.2%, respectively. CONCLUSION: No genetic toxicity of Epr was detected.     

微核试验和彗星试验检测雄黄的遗传毒性

目的用短期给药(小鼠骨髓细胞微核试验和彗星试验)和长期给药(利用生殖毒性Ⅰ段试验的大鼠做微核试验和彗星试验)检测雄黄的遗传毒性,探讨利用长期毒性试验或生殖毒性Ⅰ段试验用动物进行遗传毒性检测的可行性。方法小鼠ig给予雄黄0.25,0.5和1.0 g·kg-1,每天1次,2 d后,取骨髓细胞做微核试验和彗星试验;利用生殖毒性Ⅰ段大鼠ig给予0.125,0.25和0.55 g·kg-1,雄性连续给药42 d以上,交配成功后处死;雌性连续ig给药19 d以上,妊娠第15天,取骨髓细胞做微核试验和彗星试验,取血做外周血淋巴细胞微核试验。结果与阴性对照组比较,小鼠雄黄0.25,0.5和1.0 g·kg-1组微核试验微核率分别为3.0‰,4.40‰,7.01‰(P<0.05,P<0.01)和彗星试验拖尾率分别为6.3%,9.7%和11.3%(P<0.05,P<0.01)。与阴性对照组比较,生殖毒性Ⅰ段试验大鼠ig给予雄黄,雄黄0.55 g·kg-1组雄性大鼠的骨髓微核和外周血淋巴细胞微核率分别为2.83‰和6.67‰(P<0.05),雌性大鼠0.25和0.55 g·kg-1的骨髓微核和外周血淋巴细胞微核率分别为1.5‰,2.25‰以及2.58‰和4.40‰(P<0.05,P<0.01);雄黄使雄性和雌性大鼠彗星拖尾率明显升高(P<0.05)。结论利用生殖毒性Ⅰ段试验多次给药后取材做微核试验和彗星试验方法可行;外周血微核试验简便易行;在所观察的剂量下雄黄具有遗传毒性。     

An assessment of the genotoxicity and human health risk of topical use of kojic acid [5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one].

Kojic acid (KA), a natural substance produced by fungi or bacteria, such as Aspergillus, Penicillium or Acetobacter spp, is contained in traditional Japanese fermented foods and is used as a dermatological skin-lightening agent. High concentrations of KA (>or=1000 microg/plate) were mutagenic in S. typhimurium strains TA 98, TA 100, TA 1535, TA102 and E. coli WP2uvrA, but not in TA 1537. An Ames test following the "treat and plate" protocol was negative. A chromosome aberration test in V79 cells following a robust protocol showed only a marginal increase in chromosome aberrations at cytotoxic concentrations after prolonged (>or=18 h) exposure. No genotoxic activity was observed for hprt mutations either in mouse lymphoma or V79 cells, or in in vitro micronucleus tests in human keratinocytes or hepatocytes. All in vivo genotoxicity studies on KA doses were negative, including mouse bone marrow micronucleus tests after single or multiple doses, an in vivo/in vitro unscheduled DNA synthesis (UDS) test, or a study in the liver of the transgenic Muta(TM) Mouse. On the basis of pharmacokinetic studies in rats and in vitro absorption studies in human skin, the systemic exposure of KA in man following its topical application is estimated to be in the range of 0.03-0.06 mg/kg/day. Comparing these values with the NOAEL in oral subchronic animal studies (250 mg/kg/day), the calculated margin of safety would be 4200- to 8900-fold. Comparing human exposure with the doses that were negative for micronuclei, UDS and gene mutations in vivo, the margins of safety are 16000 to 26000-fold. In conclusion, the topical use of KA as a skin lightening agent results in minimal exposure that poses no or negligible risk of genotoxicity or toxicity to the consumer.     

降血糖活性成分Bellidifolin遗传毒性研究

目的研究降血糖活性成分Bellidifolin的遗传毒性。方法整体试验采用小鼠骨髓嗜多染红细胞微核实验；体外试验采用鼠伤寒沙门菌组氨酸营养缺陷型TA97、TA98、TA100、TA102四个菌株，对Bellidifolin进行Ames实验。结果小鼠骨髓嗜多染红细胞微核发生率结果显示Bellidifolin高、中、低剂量组与阴性对照组比较均差异无显著性(均P＞0.05)；Ames实验显示,在实验设置浓度和加S9或不加S9的实验条件下，受试物对各菌株所诱发的回变菌落数,均未超过对照的2倍。结论实验结果为阴性，未见Bellidifolin有致突变性作用。