Study On The Hapetotoxicity Mechanisim Of Traditional Uyghur Medicinal Herb Psoralea Corylifolia

Objective:To study the acute toxicity characteristics of aqueous extracts and ethanol extracts,different processed products of Fructus Psoraleae(FP)and elucidate the hapetotoxicity mechanisim of Traditional Uyghur Medicinal herb FP,and to provide reliable support for the quality control and clinical safe drug use of its related preparation.Method:(1)Study on the acute toxicity and subacute toxicity of different extracts of FP in mice by intragastric administration:18 to 22 g of 16 healthy Kunming mice were selected for each group with equal numbers of male and female mice.Maximum dosage method was used for the acute toxicity test by intragastric administration and experimental mice were randomly divided into:solvent control group and EEFP(12.0 g·kg-1)group and AEFP(18.6 g·kg-1);sub-acute toxicity test was divided into:solvent control group,0.65,1.30 g.kg-1 of EEPF group,0.8,1.6 g.k-1g of AEPF group,and after 14 days continuous administration.General observation,body weight.organ weight and coefficient,hematology,blood biochemistryand the liver,kidney histopathological examination were detected.(2)Experimental study of acute toxicity of Fructus Psoraleae with different preparation:the ethanol and aqueous extracts of different processed products(Stir Fried,Wine soaked Fried and Salt processed)and its crude powder were administrated to SPF-level mice and the acute toxical reactions and mice activities were observed in 14 days;The median lethal dose(LD50)experiment was conducted for the more virulent extracts and LD50 was calculated by using the modified Koch method.(3)Chronic hepatotoxicity mechanism of repeated administration of AEFP on rats:test was divided into 4 groups:control group,low dose group,middle dose group and high dose group.AEFP(dosage 310,600 and 1200 mg.kg-1)were administrated to SD rats for 3 months.Body weight,food intake,liver weight,serum electrolyte and biochemicals,activity of SOD,CAT,GSH-PX,MDA,NO,NOS,histopathology,gene and protein expression of FXR,CYP7A1,PPAR?,MRP3 were analyzed.Result:(1)Study on the acute toxicity and subacute toxicity of different extracts of FP in mice by intragastric administration:compared with control group,the first time after EEFP?AEFP had been administrated,mice developed the symptoms of decreasing activities,tachypnea,huddle,pilomotor,but no mice death.The second time when EEFP was administrated.14 mice were died in 40 min,rest of them recovered in 24h and the liver central venous dilatation and congestion,some glomerulus dilata-tion were found,and when AEFP was treated,apart from the above symptoms,supine,tremor,unsteady gait were occurred after 5 min and all died in 30 min and found liver hyperemia in anatomy;in sub-acute toxicity study,there were no obvious toxic symptoms and no animal death.spleen weight and coefficient of male declined and thymus weight and coefficient of female lifted obviously(P<0.05)in lowe dosage group of EEFP,female body weight increased apparently(P<0.05)and kidney coefficient dropped remarkably(P<0.01)in higher dosage group of EEFP and body weight,spleen weight,liver weight and coefficient,thymus weight lifted in high dosage group of AEFP.Hematology:neutrophil count(NEUT),monocyte(MONO)were obviously reduced(P<0.05),red blood cell(RBC),hemoglobin(HGB),hematocrit(HCT),lymphocytes(LYMPH)were obviously elevated(P<0.05)in each EEFP group of male mice;RBCwas apparently lifted(P<0.05)and platelet count(PLT),platelet hematocrit(PCT)were climbed significantly(P<0.01)in each EEPF group of female mice;RBC,HGB,HCT.LYMPH(%)of each male mice AEFP group elevated obviously or significantly(P<0.05-P<0.01);AEFP high dose male group's PCT,WBC increased significantly(P<0.05)and PLT?PCT?RBC?HGB level of high dosage female group climbed obviously or significantly(P<0.05-P<0.01).blood biochemical analysis:A/G was obviously increased(P<0.05)and bloodurea nitrogen(BUN)was apparently raised(P<0.05)in low dosage of EEFP male group;alanine aminotransferase(ALT),triglyceride(TG)were decreased significantly(P<0.01)in high-dose of EEFP male group,total bilirubin(TBIL)was ob-viously declined(P<0.05),TG was reduced remarkably(P<0.01).total bile acid(TBA),ALT were elevated obviously(P<0.05)in lower dosage and high dosage of EEFP female group.hepatocyte necrosis and glomerulus atrophy were to be found in EEFP group;TBLI elevated significantly(P<0.05),TG?GLU reduced obviously or remarkably(P<0.05,P<0.01)in low dose AEFP male group;ALB,A/G dropped apparently(P<0.05),TCHO1,GLU went down apparently or significantly(P<0.05,P<0.01)in high dose AEFP female group.AST,AST/ALT,TBA of high dosage AEFP group attenuated remarkably(P<0.05),but TCHO1 went up strikingly(P<0.01).Hepatocellular edema were to be found in AEFP.(2)Experimental study of acute toxicity of Fructus Psoraleae with different preparation:From the acute toxicity results,we can see that the aqueous extracts of Stir fried,Wine soaked fried and Salt processed FP and its crude powder expressed smaller toxicities and the maximum tolerated dose(MTD)was 50.6g.Kg-1,48.1 g.Kg-1,50.7 g.Kg-1 and 15 g.Kg-1 respectively;crude drug ethanol extracts,Stir fried ethanol extracts,Salt processed ethanol extracts and Wine soaked fried ethanol extracts of FP showed significant toxicities and LD50 of them were 36.77g.Kg-1 35.51 g.Kg-1,29.02 g.Kg-1 and 23,23 g.kg-1 for mice.The order of LD50 toxicity:Wine soaked fried ethanol extracts>Salt processed ethanol extracts>Stir fried ethanol extracts>crude drug ethanol extracts;the order of MTD toxicity:Salt processed aqueous extracts>Wine soaked fried aqueous extracts>crude powder>Stir fried aqueous extracts;The content of Bakuchiol(BAK)in ethanol extracts of different processed product was more than that of BAK in aqueous extracts of different processed products and the toxicity of ethanol extracts of different processed product was greater than their aqueous extracts.(3)Chronic hepatotoxicity mechanism of repeated administration of AEFP on rats:Abnormalities were found in AEFP-treated groups including changes of serum biochemical parameters.ascended total calcium,increased liver weight.The activity of SOD,CAT.GSN-PX were decreased,MDA and NO were went up in male rats while females were at the opposed trend to males interestingly.The mRNA and protein expression of FXR,MRP3.PPAR?.L-FABP,CYP7A1 in males were up-regulated,but FXR.PPAR? were down-regulated.MRP3 was up regulated in females.Conclusion:(1)Under the experimental condition,maximum tolerated doses(MTD)of EEFP(12.0 g.kg-1)and AEFP(18g.kg-1)were equivalent to 741 and 925 times of the clinical dosage in human respectively.EEFP(12.0 g.kg-1)can induce 70%of mice death and AEFP(18g.kg-1)can induce all mice death and indicated that it has obvious acute toxical effect.When EEFP is administrated at doses of 0.65,1.30 g.kg-1 and AEFP is administrated at doses of 0.65,1.30 g.kg-1 for 14 days in sub-acute has obvious acute toxical effect.When EEFP is administrated at doses of 0.65,1.30 g.kg-1 and AEFP is administrated at doses of 0.65,1.30 g.kg-1 for 14 days in sub-acute test,they really has certain effects on hematological,blood biochemical determinations and produce toxic abnormalities in liver and kidney.(2)The crude powder and ethanol extracts of different processed products had obvious acute toxicity and the content of BAK in ethanol extracts of different processed products was obviously higher than that of their aqueous extract and no obvious acute toxicity was observed in aqueous extracts of different processed products after administrating to mice.(3)We provided the evidence for the first time that AEFP can induce sex-related cholestatic hepatotoxicity in chronic toxicity.Females are more exposed to cholestasis and PPARa may be the main pathway for cholestasis of liver injury.