Study On The Relationship Between The Amount And Toxicity Of Toosendan Fruit Before And After Frying On The Live

Objective:Acute and chronic toxicity of extracts of Toosendan Fructus before and after stir frying were studied by acute toxicity and long-term toxicity test in mice.In order to further exploration the foundation for the mechanism of reducing poison by Toosendan Fructus.Incubation of liver microsomes and pharmacokinetics were used to investigate the effect of the activity of CYP4501A2,CYP2E1 and CYP3A4 on Toosendan Fructus and Fried Toosendan Fructus in vitro and in vivo.Thus,the mechanism of dose-toxicity change of hepatotoxicity before and after frying with Toosendan Fructus explored from the level of Internal and external molecular,which provided a guarantee for safe,reasonable and effective use of Toosendan Fructus.In order to further clarify the mechanism of the drug reduction,This paper from the perspective of absorption,the intestine absorption characteristics of the extract of Toosendan Fructus extract before and after frying were examined on Indicator component toosendanin by In the body one-way intestinal perfusion modelMethods:1.To investigate the acute toxicity of Toosendan Fructus and Fried Toosendan Fructus in mice,and calculate the median lethal dose(LD50)by Bliss.Serum liver function related biochemical markers:alanine aminotransferase(ALT),aspartate aminotransferase(AST),and the long-term toxicity of rats were investigated in combination with the pathological conditions of liver tissue;2.Using rat liver microsomal incubation in vitro with"Cocktail" probe drugs method,testing each pigment on the rat liver cells in vitro,this CYP1A2,CYP2E1 and CYP3A4 enzyme activity,and to compare its specific probe(phenacetin,chlorzoxazone and dapsone)metabolic rate;3.Use "Cocktail" probe drugs combined with pharmacokinetic test groups of rats serum this CYP1A2,CYP2E1 and CYP3A4 metabolism enzyme specificity probe,the relevant pharmacokinetic parameters are calculated by use of a DAS3.0 and draw the probe drugs-curve,AUC(0-t)and CL,t1/2 difference.4.In this paper,we used the model of rat model combined with HPLC method to determine the concentration of toosendanin in the extract before and after the preparation of Toosendan Fructus,and calculated the absorption rate constant(Ka)and the drug apparent permeability coefficient(P app)of each intestinal segment.Results:1.Study on toxicity of extracts before and after the fry of Toosendan Fructus:1.The acute toxicity was shown in the acetic acid ethyl acetate parts of fructus chinensis and stir-fry,among which the LD50 of Toosendan Fructus is 77.846g/kg,while the Fried Toosendan Fructus is 225.08 g/kg;By gavage for 90 days,Compared with the control group,serum ALT,AST,increased in all groups,Pathological examination showed different degree of liver tissue damage,and the value of biochemical indicators and pathological sections has a certain correlation with the processing and administration;2.In Vitro Study on Hepatic Cytochrome P450 Enzymes of Rats before and after Frying of Toosendan Fructus:Compared with the normal group,the metabolic rate of dapsone in the drug-administered group was higher than that of the control group after 14-day administration of Toosendan Fructus and Fried Toosendan Fructus.When the dosage of Toosendan Fructus and Fried Toosendan Fructus was the same,the metabolic rate of dapsone in the Toosendan Fructus group was higher than that of the Fried Toosendan Fructus.Under the same processed products,the high-dose group was larger than the middle dose group and the low dose group on the metabolic rate of dapsone.However,compared with the control group,whether given the Toosendan Fructus and Fried Toosendan Fructus the metabolic rate was not statistically significant on phenacetin and chlorzoxazone.3.In Vivo Study on Hepatic Cytochrome P450 Enzymes of Rats before and after Frying of Toosendan Fructus:Compared with the normal group,the AUC of phenacetin and dapsone in different groups was lower than that in the normal group and the CL value was increased after the rats were given Toosendan Fructus and Fried Toosendan Fructus for 14 days.In the same dose,the AUC value of Fried Toosendan Fructus was higher than that of Toosendan Fructus and the CL value was lower than that of Toosendan Fructus.The highest dose of AUC was greater than the low dose group of the same processed products.And the AUC and CL of chlorzoxazone were no different from the normal group;4.Effect of Toosendan Fructus before and after Sowing on Intestinal Absorption in Rats:Toosendanin in the rat duodenum,jejunum,ileum,colon absorption rate constant(Ka)in turn were raw extract 746.30±40.31/s～347.84±111.67/s,fried Toosendan Fructus extract in the range of 473.77±22.05/s～243.33±19.58/s,Toosendanin in each segment of the rat intestine of the drug apparent permeability coefficient(Papp)followed by the crude extract 83.66±7.00 cm/s～58.33±16.49cm/s,fried Toosendan Fructus extract 43.57±8.63cm/s～33.89±1.55cm/s.Conclusion:1.The acute toxicity and long-term toxicity of rats showed that stir frying could reduce the toxicity and dose dependence of Toosendan Fructus,which laid the foundation for further study on the reduction of the toxicity of stir frying;2.In vitro metabolism study of rat liver microsomes showed that Toosendan Fructus can induce the activity of CYP3A4,but has no significant effect on CYP1A2 or CYP2E1.The activity of Toosendan Fructus induced CYP3A4 may be related to the dose,and the reason for stir-frying to reduce the toxicity may be related to the inhibition of the induction of CYP3A4 after stir-frying.In this way,the mechanism of detoxification was preliminarily analyzed from the point of view of in vitro;3.According to the pharmacokinetic study of rats in vivo,the AUC of different groups of nalazine and sulfoxide in Toosendan Fructus were lower than that of the normal group and the CL value increased,and the AUC value of the same dose was higher than that of Toosendan Fructus and the CL value was lower than that of the raw Toosendan Fructus;the AUC value of the high dose of the same product was greater than the low dose group,and the AUC and CL of the chlorzoxazone were no difference compared with the normal group;The results showed that Toosendan Fructus can induce the activity of CYP1A2 and CYP3A4,and has no induction or inhibition effect on CYP2E1.The mechanism of attenuating by stir-fryring may be related to the lowering of inducing effect of Toosendan Fructus.In this way,the mechanism of detoxification was preliminarily analyzed from the point of view of vivo;4.According to the study of intestinal absorption characteristics showed that the index component toosendanin of Toosendan Fructus in the intestinal absorption is good.There is a difference in the absorption in different intestinal segments on Toosendan Fructus and Fried Toosendan Fructus.It is suggested that stir-frying may affect the intestinal rate and the apparent permeability of toosendanin which is the index component of Toosendan Fructus.Thus,from the perspective of intestinal absorption,we preliminarily reveal the mechanism of Toosendan Fructus reducing toxicity by stir frying.