I. C. S. Carvalho, T. P. Dutra, Dennia Perez de Andrade, I. Balducci, C. Pacheco-Soares, R. F. Rocha
{"title":"怀孕期间高剂量的酒精会导致新生大鼠成骨细胞的DNA损伤。","authors":"I. C. S. Carvalho, T. P. Dutra, Dennia Perez de Andrade, I. Balducci, C. Pacheco-Soares, R. F. Rocha","doi":"10.1002/bdra.23468","DOIUrl":null,"url":null,"abstract":"BACKGROUND\nAlcohol exerts teratogenic effects and its consumption during pregnancy can cause deficit of bone development. The aim of the current study was to evaluate the genotoxic effects of prenatal exposure to ethanol on newborn rat osteoblasts.\n\n\nMETHODS\nWistar rats were initially divided into two groups: Ethanol group which received Ethanol 20% V/V in liquid diet and solid diet ad libitum, and Control group, which received solid diet and water ad libitum. Each group received a specific diet for 8 weeks before breeding and throughout three weeks of gestation and the treatment was finished on the day the pups were killed. On the fifth day of life, the pups from each group were killed for removal of the calvaria and isolation of osteogenic cells by sequential enzymatic digestion. The cells were cultured for a maximum period of 14 days. The detection of genotoxic effects of alcohol was investigated by the comet and the micronucleus assay.\n\n\nRESULTS\nMicronucleus and comet assay showed significant increases in DNA damage at 7 days in Ethanol group (p = 0.0302, p = 0.0446, respectively). However, at 14 days both assay showed no significant difference between the groups (p = 0.6194, p = 0.8326, respectively).\n\n\nCONCLUSION\nOur results showed that prenatal exposure to ethanol induced DNA damage in osteoblasts, as shown by micronucleus formation and higher percentage of DNA in the comet tail. It can be concluded that prenatal exposure to ethanol damages osteoblast DNA in newborns exposed to high doses of ethanol during pregnancy, suggesting that prenatal ethanol consumption has a direct effect on fetal osteoblasts.","PeriodicalId":8983,"journal":{"name":"Birth defects research. Part A, Clinical and molecular teratology","volume":"121 1","pages":"122-32"},"PeriodicalIF":0.0000,"publicationDate":"2016-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"14","resultStr":"{\"title\":\"High doses of alcohol during pregnancy cause DNA damages in osteoblasts of newborns rats.\",\"authors\":\"I. C. S. Carvalho, T. P. Dutra, Dennia Perez de Andrade, I. Balducci, C. Pacheco-Soares, R. F. Rocha\",\"doi\":\"10.1002/bdra.23468\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"BACKGROUND\\nAlcohol exerts teratogenic effects and its consumption during pregnancy can cause deficit of bone development. The aim of the current study was to evaluate the genotoxic effects of prenatal exposure to ethanol on newborn rat osteoblasts.\\n\\n\\nMETHODS\\nWistar rats were initially divided into two groups: Ethanol group which received Ethanol 20% V/V in liquid diet and solid diet ad libitum, and Control group, which received solid diet and water ad libitum. Each group received a specific diet for 8 weeks before breeding and throughout three weeks of gestation and the treatment was finished on the day the pups were killed. On the fifth day of life, the pups from each group were killed for removal of the calvaria and isolation of osteogenic cells by sequential enzymatic digestion. The cells were cultured for a maximum period of 14 days. The detection of genotoxic effects of alcohol was investigated by the comet and the micronucleus assay.\\n\\n\\nRESULTS\\nMicronucleus and comet assay showed significant increases in DNA damage at 7 days in Ethanol group (p = 0.0302, p = 0.0446, respectively). 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引用次数: 14
摘要
背景:酒精具有致畸作用,怀孕期间饮酒可导致骨骼发育缺陷。本研究的目的是评估产前暴露于乙醇对新生大鼠成骨细胞的遗传毒性作用。方法将swstar大鼠初始分为两组:乙醇组,在液体和固体饲粮中随意添加20% V/V的乙醇;对照组,在固体饲粮中随意添加水。每组在繁殖前8周和妊娠3周内给予特定的饮食,并在幼崽被杀死当天结束治疗。在出生第5天,每组幼鼠处死,通过顺序酶消化去除颅骨和分离成骨细胞。细胞最多培养14天。采用彗星法和微核法研究了酒精的遗传毒性作用。结果7 d时,乙醇组小鼠DNA损伤显著增加(p = 0.0302, p = 0.0446)。但在第14天,两组间均无显著差异(p = 0.6194, p = 0.8326)。结论产前乙醇暴露可诱导成骨细胞DNA损伤,表现为微核的形成和彗尾中较高的DNA百分比。由此可见,胎儿在孕期暴露于高剂量乙醇环境下,其胎儿的成骨细胞DNA受到乙醇的破坏,提示胎儿在孕期摄入乙醇对胎儿成骨细胞有直接影响。
High doses of alcohol during pregnancy cause DNA damages in osteoblasts of newborns rats.
BACKGROUND
Alcohol exerts teratogenic effects and its consumption during pregnancy can cause deficit of bone development. The aim of the current study was to evaluate the genotoxic effects of prenatal exposure to ethanol on newborn rat osteoblasts.
METHODS
Wistar rats were initially divided into two groups: Ethanol group which received Ethanol 20% V/V in liquid diet and solid diet ad libitum, and Control group, which received solid diet and water ad libitum. Each group received a specific diet for 8 weeks before breeding and throughout three weeks of gestation and the treatment was finished on the day the pups were killed. On the fifth day of life, the pups from each group were killed for removal of the calvaria and isolation of osteogenic cells by sequential enzymatic digestion. The cells were cultured for a maximum period of 14 days. The detection of genotoxic effects of alcohol was investigated by the comet and the micronucleus assay.
RESULTS
Micronucleus and comet assay showed significant increases in DNA damage at 7 days in Ethanol group (p = 0.0302, p = 0.0446, respectively). However, at 14 days both assay showed no significant difference between the groups (p = 0.6194, p = 0.8326, respectively).
CONCLUSION
Our results showed that prenatal exposure to ethanol induced DNA damage in osteoblasts, as shown by micronucleus formation and higher percentage of DNA in the comet tail. It can be concluded that prenatal exposure to ethanol damages osteoblast DNA in newborns exposed to high doses of ethanol during pregnancy, suggesting that prenatal ethanol consumption has a direct effect on fetal osteoblasts.