Based On The Jaundice And TK-TD Model To Study The "Quantity-Time-toxic" Relationship And The Basis Of Toxic Substances In Vivo Of Gardenia Hepatotoxicity

Objective:In this experiment,the amount-time-toxicity relationship of the gardenia extract was determined by single-time and multiple-dose administration of the gardenia extract,and the amount-toxic curve of the liver-renal toxicity of the gardenia extract was determined.Dose differences in toxicity were observed in rats with jaundice.The toxicokinetic-toxic kinetics?TK-TD?binding model study of geniposide and its metabolites clarifies the concentration-effect-time relationship and clarifies the material basis of hepatotoxicity induced by Chinese medicine-gardenia.Methods:1.Using 0.48,0.76,1.16,1.8,2.76,4.26 g·kg^-1gardenia extracts to study the quantitative-toxic relationship between normal rats and jaundice model rats.Blood was taken after 24 hours of single administration.Liver and kidney function tests:total bilirubin?TBIL?,alanine aminotransferase?ALT?,aspartate aminotransferase?AST?,alkaline phosphatase?ALP?,urea nitrogen?BUN?,creatinine?CREA?and pathological anatomy to determine the amount-toxic relationship of scorpion extract to liver and kidney toxicity.2.Three doses of gardenia extract(0.14,0.28,0.56 g·kg^-1)were administered orally to rats for 4 weeks,and the liver and kidney function of rats were detected and killed at 2and 4 weeks respectively.Some animals underwent liver and kidney pathological examina-tion,and some animals were reserved for stopping the drug and observing whether the toxicity was reversible?observation period 2 weeks?.3.TK study was performed in normal animals and scutellaria animals.After admini-stration,blood was taken to separate plasma,blood concentration-time curve was drawn,and relevant pharmacokinetic parameters were calculated.The DAS pharmacokinetic software was used to calculate the pharmacokinetic parameters.The plasma concentration of geniposide and genipin was used as the TK index,the activity of ALT,AST and TBIL was used as the TD index,and the TK-TD combination model was proposed by DAS software.And for the analysis of geniposide metabolites in plasma.Results:1.Compared with the blank group,ALT,AST and TBIL were significantly increased in the dose groups of 1.16,1.8,2.76 and 4.26 g·kg^-11 in normal rats,and were positively correlated with the dose?P<0.05?.Compared with the control group of the jaundice model,the doses of ALT,AST,ALP and TBIL were significantly increased in the dose groups of 0.76,1.16,1.8,2.76 and 4.26 g·kg^-1?P<0.05?.Compared with the blank group,the BUN and CREA of the rats in the 0.48,0.76,1.16,1.8,2.76,4.26 g·kg^-11 dose groups increased in different degrees?P<0.05,P<0.01?.BUN and CREA increased in different doses in the 0.76,1.16,1.8,2.76,and 4.26 g·kg-1 dose groups?P<0.05,P<0.01?,and showed a signifi-cant dose with a certain range.Upward trend;2.After 14 days of administration,ALP,AST and TBIL were significantly increased in the 0.28 and 0.56 g·kg^-11 dose groups compared with the model control group?P<0.05,P<0.01?.After 28 days of administration,compared with the blank group,the TBIL of the normal 0.56 g·kg-1 dose group was significantly increased?P<0.05?.Compared with the model control group,ALP and AST were significantly increased in the 0.14 g·kg^-11 dose group?P<0.05,P<0.01?,and the model was administered in the 0.28 g·kg^-11 dose group.TBIL was significantly increased?P<0.05?.After 2 weeks of drug withdrawal,there was no significant difference in ALP,ALT,AST and TBIL between the groups.3.Geniposide 1.2 g·kg^-11 was administered by gavage,and the linear relationship between geniposide and genipin was 72.25-14450 ng·mL^-1and 45.6-9120 ng·mL^-1.The endogenous substances in the plasma do not interfere with the determination of the components,and the method meets the measurement requirements.The mean plasma concentration-time curve of geniposide showed a bimodal phenomenon.The toxicokinetic parameters were calculated using a non-compartmental model.The area under the curve of the model group was AUC?0-t?and the peak concentration(C_max)and average.The residence time?MRT?was significantly higher than the normal group and the half-life was prolonged.The concentration of genipin in the model group was higher than that in the normal group.A significant counterclockwise hysteresis loop can be seen in the toxicolo-gical effect-concentration curve.The TK-TD model conforms to the Sigmoid-E_maxax model.Sixteen metabolites were identified in plasma,and two groups of differential metabolites were G2,6,11.Conclusion:1.The dose of gardeniat extract exceeding 1.16 g·kg^-1has obvious acute hepatotoxi-city in normal rats,and it is dose-dependent.The dose of gardenia extract exceeding 0.76g·kg^-11 can cause acute liver injury in rats with jaundice,and the dose of 0.76-4.26 g·kg^-1can cause more serious liver damage to the jaundice model.The toxic dose of the rats with jaundice was lower than that of normal rats.The dose of 0.76-4.26 g·kg^-11 of gardenia extract caused significant kidney damage in both normal rats and jaundice model rats in a dose-dependent manner.The 1.16-4.26 g·kg^-1dose caused more severe kidney damage in the model of jaundice.2.The dose of gardenia extract is more than 0.28 g·kg^-11 for 14 days,which can cause certain liver damage to rats with jaundice.Large doses of gardenia extract 0.56 g·kg^-1for28 days of continuous administration caused certain liver damage in normal rats.The dosage of gardenia extract was less than 0.28 g·kg^-11 for 28 days.There was no obvious liver injury in normal rats.With the prolongation of administration time,the dose was greater than 0.14 g·kg^-11 for 28 days.Model rats caused significant liver damage.After 2weeks of drug withdrawal,there was no significant difference in liver and kidney indexes between the groups,indicating that the dose of gardenia extract was reversible in the liver and kidney damage caused by normal and jaundice rats at doses of 0.14⁰.56 g·kg^-1.3.After intragastric administration of geniposide,the concentration of jaundice model rats was higher than that of normal rats,and the elimination was slow and the residence time was longer.It indicated that geniposide and genipin accumulated severely in the model of jaundice membranaceus,and the duration was long.The toxicological effect lags behind the blood drug concentration,and there may be hepatic and intestinal circulation.Combined with previous experiments,it is speculated that geniposide is the material basis for the toxicity of gardenia,and the direct toxic substance is based on the in vivo metabolite of geniposide,genipin.