| Objective:To analyse the main components in water extract of Polygoni Multiflori Radix(PMR)from Sichuan and Hubei province.To investigate the hepatotoxicity of water extract of PMR on normal rats and rats with hepatic cytochrome P450 enzyme(CYP450)activity inhibited.Methods:1.HPLC was employed to analyze the main components in water extract of PMR.2.To investigate the hepatotoxicity of water extract of PMR on rats for oral administration continuously:forty-eight SD rats were randomly divided into 6 groups:control group,PMR group,CYP1A2 inhibited group,CYP2E1 inhibited group,PMR+CYP1A2 inhibited group,PMR+CYP2E1 inhibited group.The rats in CYP1A2 inhibited group and PMR+CYP1A2 inhibited group were received an intraperitoneal(i.p.)injection of CYP1A2 inhibitor(cimetidine)of 50 mg·kg-1 BW at 5 days prior to the oral administration.The rats in CYP2E1 inhibited group and PMR+CYP2E1 inhibited group were received an intraperitoneal(i.p.)injection of CYP2E1 inhibitor(trans-1,2-dichloroethylene)of 100 mg·kg-1 BW at 2.5 h prior to the oral administration.The rats in PMR group,PMR+CYP1A2 inhibited group and PMR+CYP2E1 inhibited group were received an oral administration of PMR of 30g·kg-1 body weight(BW).On the 2 h after 1 d,8 d,15 d,22 d,28 d with PMR treatment,the blood samples from rats were collected.Alanine aminotransferase(ALT),aspartate aminotransferase(AST)and alkaline phosphatase(ALP)were tested.3.To investigate the hepatotoxicity of different dose of water extract of PMR on normal rats for oral administration continuously and interval:rats were given 10 g·kg-1 BW water extract of PMR for 14 d,interval 14 d,then 1 d,interval 14 d,then 1 d,interval 7 d,and then 1 d.The rats were draw blood in the 1 d,7 d,29 d,37 d lavage after 2 h and 2 d,14 d,37 d lavage after 24 h,respectively.ALT,AST,ALP,TBIL(total bilirubin),direct bilirubin(DBIL),total protein(TP),albumin(Alb),total cholesterol(TCHO)were tested.Pathological examination of liver were performed.4.To investigate the hepatotoxicity of water extract of PMR on CYP1A2 inhibited rats for oral administration:forty-eight SD rats were randomly divided into 6 groups:control group,PMR low dose group,PMR high dose group,CYP1A2 inhibited group,CYP1A2 inhibited+PMR low dose group,CYP1A2 inhibited+PMR high dose group.The rats were received an oral administration of PMR of 10g·kg-1 BW and 30g·kg-1 BW for 7 d,continuously.The rats were draw blood in the 1 d,7 d lavage after 2 h and 2 d lavage after 24 h,respectively.The hematologic and biochemical indices were tested.5.To investigate the hepatotoxicity of water extract of PMR on CYP2E1 inhibited rats for oral administration:sixty SD rats were randomly divided into 6 groups:control group,PMR low dose group,PMR high dose group,CYP2E1 inhibited group,CYP2E1 inhibited+PMR low dose group,CYP2E1 inhibited+PMR high dose group.The rats were received an oral administration of PMR of 10g·kg-1 BW and 30g·kg-1 BW for 14 d,continuously.The rats were draw blood in the 1 d,7 d and 14 d lavage after 2 h and 24 h,respectively.The hematologic and biochemical indices were tested.Results:1.The HPLC analysis results show that the main components in water extract of PMR from Hubei has 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glycosidase(short for TSG),emodin-8-O-β-D-glucoside,emodin,emodin methyl ether.The component was(145.41±4.28)μg·mL-1,(13.56±0.20)μg·mL-1,(1.37±0.12)μg·mL-1,(0.45±0.02)μg·mL-1,respectively.The component in PMR from Sichuan was same with PMR from Hubei.The component was(49.04±4.73)μg·mL-1,(6.84±0.16)μg·mL-1,(0.91±0.09)μg·mL-1,(0.31±0.01)μg·mL-1.2.Normal rats were given 10 g·kg-1·BW water extract of PMR for 28 d continuously,the experimental group weight decreased obviously,feeding rate has no obvious change,the ALT,AST and ALP of rats had no obvious effect,but that rats were found that tired,stool soft and so on.The rats with activity of CYP1A2,CYP2E1 enzyme inhibited,the ALT,AST and ALP were no significant for 28 d lavage continuously.3.Continuously and interval intragastric administration of 10 g·kg-1·BW water extract of PMR,continuously administration for 14 d.The TBIL and IBIL was increased significantly than control group for 7 d after 2h.TBIL and DBIL in rat had an increased in 7d 2 h.ALT were significantly increased for interval after 14 days of administration 1d 2 h and at an interval of 7 d administration 1d 2 h.But ALT had no increase significantly for an interval of 7 d administration 1d 24 h.Liver pathology analysis also found no obvious damage of rat liver.4.Normal rats were given 10 g·kg-1·BW,30 g·kg-1·BW water extract of PMR for 7 d.The ALT of rats with high dose compared with the control group were significantly increased for 1 d 2 h,2 d 24 h and 7 d 2 h;compared with the PMR low dose group were significantly increased for 7 d 2 h.The TBIL and IBIL of rats with high dose compared with the control group were significantly increased for 1 d 2 h and 7 d 24 h.The rats with the activity of CYP1A2 inhibited were given high dose of water extract of PMR,the AST and TBIL were significantly increased for 2 d 24 h.But compared rats with 30 g·kg-1·BW water extract of PMR,there was no significant difference.5.Normal rats and rats with inhibited CYP2E1 enzyme activity were given 10 g·kg-1·BW,30 g·kg-1·BW water extract of PMR for 14 d inhibiting.The results showed that water extract of PMR can make an influence on the CYP2E1 enzyme activity inhibited in the rats of blood biochemical parameters,can change the level of TBIL,DBIL and IBIL.Liver biopsy showed that vacuole hepatic cells and fibroblast proliferation in normal rats with 30 g·kg-1·BW water extract of PMR.TBIL and IBIL in rats with CYP2E1 inhibited had a significantly increase with given 30 g·kg-1·BW PMR for 2 h after.Conclusion:1.There were a significant differents between two sources of PMR in main components.The components of PMR from Hubei was higher than PMR from Sichuan.2.The rat with 10 g·kg-1·BW water extract of PMR had no significantly increased in ALT,AST and ALP.30 g·kg-1·BW water extract of PMR can cause abnormal liver function of rats in a short time.High dose PMR had a potential risk of liver toxicity in clinical.3.10 g·kg-1·BW PMR had no significant effect on CYP1A2 and CYP2E1 enzyme activity inhibition of rat in transaminase levels,but can be caused by inhibition of CYP1A2 enzyme activity of rat blood TBIL and IBIL increased significantly,30 g·kg-1·BW PMR can be caused by inhibition of CYP1A2,CYP2E1 enzyme activity of rat blood TBIL increased significantly.Therefore,liver toxicity of PMR can obviously with the low level of CYP2E1 or CYP1A2. |
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