Regulative Effect Of T-2 Toxin And The Conbination With T-2 And Deoxynivalenol On Rats Liver Microsomal CYP1A2

ObjectiveTo observe the effects of T-2 toxin and T-2 combined with deoxynivalenol on Liver Microsomal CYP1A2 of rats in growth period,to explore the pathogenesis and united pathogenic effect.And observe the relationship between histopathology examination and the characteristics,to provide the technological approaches for the further study of the prevention of mycotoxin poisoning.Methods1.Thirty healthy Wistar male rats(28days)divided randomly into 3 groups:control,T-2 toxin,‘T-2 toxin+DON toxin group’,conduct a short-term toxicology experiment,the rats of T-2 dosage treatment group were taken orally administered with 150μg/kg;the rats of ‘T-2 toxin and DON toxin’ dosage treatment groups were taken orally administered 100μg/kg and 220μg/kg everyday.The experimental period was divided into 15 days and 30 days.The livers were quickly dissected after the rats were decapitated,prepared the rat liver microsomes by using differential ultracentrifugation,then put microsomes in the-80℃ fridge.2.Tested the change of content and activity of liver microsomal CYP1A2 of rats by the means of ELISA and observe the toxic influence of T-2 toxin and ‘T-2 toxin + DON toxin’ on livers.Cut the tissues of the livers,the colons,the kidneys and the third esophageal stenosises of the rats to make paraffin sections,and HE staining was used for to test pathological changes of each organs above.Results1.The content of CYP1A2 was 5.560±0.287(ng/ml)and 3.825±0.120(ng/ml)in the control group and ‘T-2 toxin’ group after fifteen days’ contamination,the two groups were compared with independent t test,P<0.01.2.The second period lasts thirty days,the content of CYP1A2 was 7.300±0.687(ng/ml),3.224±0.318(ng/ml),2.254±0.133(ng/ml)in the control group,‘T-2 toxin ’group and the data of united group were analyzed with one-way ANOVA,compared the one and one with LSD and Dunnett methods,the results are P<0.01.The activity of three groups was 14.026±1.17(U/ml),9.358±0.509(U/ml),8.89±0.727(U/ml),the three groups of data were analyzed by one-way ANOVA procedure and LSD and Dunnett methods,compared with control group,T-2 toxin and the united toxin with T-2 and DON significantly inhibit the activity of CYP1A2(P<0.01),the results between T-2 and ‘T-2 toxin + DON toxin’ was no significant difference in the activity of CYP1A2(P>0.05).3.When the expose time lasts 15 days,the content of liver microsomal CYP1A2 is 3.825±0.12(ng/ml);when the expose time lasts 30 days,the content of liver microsomal CYP1A2 is 3.224±0.318(ng/ml).Test the two period of the content of liver microsomal CYP1A2 through the two-independent-samples t-test,the result is t=3.99,P=0.011(P≤0.05),it can illustrate that the content of rats’ liver microsomal CYP1A2 will changes when the expose time varies from 15 days to 30 days.4.The tissues of histopathological examinations have presented that,compared with the normal organs for the control group,there is no obvious pathological changes among the livers for rats after 15 days;while part of hepatic cells edema and irregular vacuole could be found in the group of ‘T-2 toxin’ after 30 days;and in ‘T-2 toxin + DON toxin’ group,it showed significant enlargement of liver cells and plasma osteoporosis and ballooning changes.The intestinal mucosal of the group of ‘T-2 toxin’ rats become edematous,and the gap was increased,the mucosal layer was appeared a number of inflammatory cellular infilitrations after 15 days;while it shows that some epithelial cells of the intestines mucous were denaturation,necrosis and defluvium defluxion in the ‘T-2 toxin’ group after 30 days;T-2 toxin + DON toxin group shows that epithelial cells of the intestines mucous were denaturation,necrosis and defluvium defluxion,the fixed mucous layer edematous and the gap between cells increased.The protein casts were founded for the kidneys in the rats of the ‘T-2 toxin’ group,there are leaking pink protein fluid during the lumenof the renal tubules and the epithelial cells of renal tubular edema,and there are also irregular and obscured boundary spaces in cytoplasm after 15 days;while there exist a mesangial proliferation as well as the vanished capsular space of the glomeruli in ‘T-2 toxin’ group rats after 30 days;The ‘T-2 toxin + DON toxin’ group shows the protein casts,there are leaking pink protein fluid during the lumenof the renal tubules and pink dropping liquid could bu found in the capsular space of the glomeruli.The tissue appearance of esophagus of the ‘T-2 toxin’ group are normal after 15 days;while histopathological changes of esophageal epithelium become incrassation and hyperkeratosis after 30 days in the ‘T-2 toxin’ group;The ‘T-2 toxin + DON toxin’ shows histopathological changes of esophageal epithelium become incrassation and hyperkeratosis and hyperplasia of squamous epithelium.Conclusions(1)T-2 toxin can lead to the content and activity of CYP1A2 both decreased,and the combined effects with ‘T-2 toxin + DON toxin’ were aggravated.This might be one of the mechanisms of which the diseases induced by T-2 toxin and combined toxin.(2)T-2 toxin can lead to the pathological injures of the livers,the the colons,the kidneys and the esophagi,and the combined effects of ‘ T-2 toxin + DON toxin ’ were aggravated.(3)The exposed time has a influence on the content and the activity of the rats’ liver microsomal CYP1A2,and with the exposed time getting longer and longer,the content and the activity of the rats’ liver microsomal CYP1A2 both decreased,and the damage to the rats’ tissues and organs become more severe.