| Sodium dehydroacetate(DHA-S)is widely used in food,feed,cosmetics and other fields as a food additive and fungicide.However,DHA-S was found cause coagulation disorders in rats,and the effect was greater on female than on male rats.The concentration of DHA-S in female blood was significantly higher than that in male rats.The obvious sex difference was found in the coagulation abnormal induced by DHA-S and its blood distribution.In our previous research,DHA-S was found induce coagulation disorders in broilers,which can significantly prolong the coagulation indexes prothrombin time(PT)and activated partial prothrombin time(APTT)of chickens.The concentration of DHA-S in the blood and liver of female chickens were higher than that of male chickens.It is well known the drug effect is directly related to its blood concentration in the body,and the blood concentration level is inseparable from the body’s metabolism of the drug.In this study,DHA-S was oral administrated to rats and chickens,and the DHA-S blood concentrations were determined by HPLC method.The pharmacokinetic characteristics of DHA-S in rats and chickens were analyzed by pharmacokinetic software.ELISA method and Cocktail probe method were used to study the effects of DHA-S on the CYP450 enzyme in rat liver,and to explore the difference of the effects of DHA-S on the coagulation function of chickens and rats as well as the sex differences.It will be benefit to the rational application of DHA-S in different animal feed and to prevention of the side effect on coagulation function of animals in clinical application.In the pharmacokinetic studies of DHA-S on rats and broilers,blood samples were collected at 0,0.083,0.25,0.5,1,2,3,5,7,9,11,24,36,48,72 h after 150 mg/kg DHA-S was given rats by gavage.The chicken blood samples were collected at 0,0.33,0.66,1,2,3,4,6,8,10,12,24,36,48,and 60 h after broilers were given DHA-S at 150 mg/kg and 200 mg/kg DHA-S by gavage.The content of DHA-S in plasma was determined by HPLC method established in our laboratory and the pharmacokinetic parameters were analyzed by Winnolin 6.3 pharmacokinetic software.The results showed that the pharmacokinetic characteristics of DHA-S in 150 mg/kg DHA-S administration to rats were as follows:the peak time was 0.85～1.0 h,the peak concentration was 322～406 mg/L.There were gender differences in rat pharmacokinetic parameters such as T1/2β,Cmax,AUC0-∞,and MRT.The T1/2β,Cmax,AUC0-∞,and MRT of DHA-S in female rats were significantly higher than those in male rats.The elimination of DHA-S in female rats is slower.After 150 mg/kg and 200 mg/kg DHA-S were given to broilers,the peak time was 4～10 h,and the peak concentration was about 200 mg/L and 300 mg/L,respectively.T1/2β,Tmax,Cmax,AUC0-∞ and MRT of female chickens were higher than those of male chickens at 150 mg/kg or 200 mg/kg treatments,but no significant differences.There was no significant sex difference in the pharmacokinetic parameters of DHA-S in chickens.The pharmacokinetics of DHA-S in rats was found different from that in chickens,which the absorption rate and extent of DHA-S in chickens were lower than those in rats,there was no significant difference in the elimination of DHA-S in chickens and rats.In the study of DHA-S on the rats CYP450 enzyme,the enzyme contents and mRNA levels of CYP450 subenzymes in liver were determined by ELISA and RT-PCR after 150 mg/kg DHA-S was administered for 5 days.The results showed that the CYP3A1,CYP1A2,CYP2C11,CYP2E1 enzyme contents and mRNA levels in the liver of the DHA-S treatment were not significantly different from those of control group.The Cocktail probe method was used to study the effect of DHA-S on CYP450 enzyme in rat liver,with dapsone,finacetine,omeprazole and clozoxazone as the probe substrates for CYP3A1,CYP1A2,CYP2C11 and CYP2E1,respectively.A HPLC method for the simultaneous determination of the four probe drugs in rat plasma was established.The chromatographic conditions were C18 column,graded elution of acetonitrile and 0.02 mM phosphate buffer(pH 5.8),and detected at 245 nm,280 nm,293 nm and 302 nm wavelengths respectively.The accuracy,precision and sensitivity of the proposed method were validated for the requirements of quantitative analysis.After continuous administration of 150 mg/kg DHA-S for 5 days,rats were given mixed probe drugs by gavage.Blood samples were collected at different times,and the concentration of probe drugs in plasma was determined by HPLC.Results showed as follows.In the pharmacokinetic parameters of finacetine probe,the Tmax,Cmax,AUC0-∞ of female rats in DHA-S group were significantly higher than those in the control group,and Vz and CL were significantly decreased,while there was no significant difference in the pharmacokinetic parameters of male rats between DHA-S group and the control group.In the pharmacokinetic parameters of omeprazole probe,the T1/2β,Tmax,AUC0-∞ and MRT of female rats in DHA-S group were significantly higher than those of control group,and the CL was significantly reduced.The MRT of male rats was significantly higher than that of control group.In the pharmacokinetic parameters of clozoxazone probe,there was no statistical difference in the parameters of female and male rats between the DHA-S group and the control.In the pharmacokinetic parameters of dapsone probe,the AUC0-∞ of female rats in DHA-S experimental group was significantly higher than that of control group,and the CL was significantly lower,while the CL value in male rats was significantly lower than that of control group.According to the pharmacokinetic parameters of each probe drug,DHA-S was found significant inhibite the CYP1A2,CPY2C11 and CYP3A1 enzymes in rats,with greater inhibitory in female rats than in male rats.The CYP2E1 activity in rats was not significant inhibited by DHA-S.In summary,there are differences in the pharmacokinetics of DHA-S between rats and chickens:the sex differences in the pharmacokinetic parameters of DHA-S in rats was obvious found;there was no sex differences in the pharmacokinetic parameters of DHA-S in broilers.The enzymes of CYP1 A2,CPY2C11 and CYP3A1 in rat liver were significantly inhibited by DHA-S with more inhibitory effect in female rats than in male rats;the liver CYP2E1 enzyme in rats may be not significantly affected by DHA-S. |
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