Toxicity effects of CCl4 on DNA and expression of CYP3A gene in hepatocytes of Cyprinus carpio var.Jian
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Key Open Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology,Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Science,Key Open Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology,Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Science,Key Open Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology,Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Science,Key Open Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology,Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Science,Key Open Laboratory for Genetic Breeding of Aquatic Animals and Aquaculture Biology,Ministry of Agriculture,Freshwater Fisheries Research Center,Chinese Academy of Fishery Science

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    Abstract:

    Carbon tetrachloride(CCl4)has been widely used as a kind of classic liver poison in constructing mammalian liver injury model and screening liver protecting drug.To investigate toxicity effects of CCl4 on hepatocyte DNA of Cyprinus carpio var.Jian,the alkaline single-cell gel electrophoresis(comet assay),histopathological examination of liver and the measuring of biomarkers of hepatocyte damage were performed respectively after low,middle and high concentrations(6.25% and 12.50% and 15.00% CCl4 in olive oil,v/v)of CCl4 at a volume of 0.01 mL/g of body weight being injected intraperitoneally into C.carpio var.Jian for 72 h.Meanwhile,the Real-time PCR was carried out to elevate the effects of CCl4 on CYP3A mRNA expression.Results showed that the group injected with 6.25% of CCl4 for 72 h has no obvious change(P>0.05)on histopathological examination,serum hepatase activities and mRNA expression of CYP3A,compared to the control,however,the comet assay results of hepatic cells showed that tail length,tail DNA(%),tail moment and olive tail moment increased significantly(P<0.01).With the increase of concentrations,histological changes such as cell swelling,extensive vacuoles degeneration,karyopyknosis and karyolysis were observed.The groups injected with 12.5% and 15% of CCl4 could significantly elevate the serum levels of glutamate oxalate transaminase(GOT)and glutamate pyruvate transaminase(GPT)(P<0.01 or P<0.05),and the group with 15.00% of CCl4 produced a significant effect in the case of lactate dehydrogenas(LDH)and malondialdehyde(MDA)(P<0.05).All damage indexes of comet assay also increased with the increase of CCl4 concentrations(P<0.01).At the same time,the expressions of CYP3A of the groups injected with 12.5% and 15% of CCl4 were significantly reduced(P<0.05).Overall results proved the toxicity effects of CCl4 on hepatocyte DNA of C.carpio var.Jian.Within the range of experimental concentrations,CCl4 could inhibit CYP3A mRNA expression.And the results of serum hepatase activities,histopathological examination and comet assay were observed not only in a dose-dependent manner but also having good coherence with the increase of CCl4 concentrations.The technique of comet assay showed higherr sensitivity.

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CAO Liping, JIA Rui, DU Jinliang, DING Weidong, YIN Guojun. Toxicity effects of CCl4 on DNA and expression of CYP3A gene in hepatocytes of Cyprinus carpio var. Jian[J]. Journal of Fisheries of China,2013,37(10):1452~1459

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History
  • Received:May 28,2013
  • Revised:August 28,2013
  • Adopted:October 16,2013
  • Online: October 25,2013
  • Published:
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