The Effect Of Polygonum Multiflorum On Liver Toxicity

Objective: This paper aims to observe the effects of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata on hepatotoxicity at the cellular level and animal level,to evaluate the potential liver injury induced by Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata and its hepatoxic compounds.Methods:(1)Cell viability was determined by MTT in vitro.Ten mainly known compounds in Polygonum multiflorum(stilbene glycoside,polydatin,aloe emodin,Rhaponticin,emodin,chrysophanol,rhein,Physcion,Hyperoside,gallic acid)were slected,and their inhibition on L-02 cell viability was observed at the concentration of 100?M.The compounds with most inhibition were further observed with different concnetrations(1,10,25,50,100?M)on cell viability in L-02 cells.(2)Acute toxicity experiment in vivo:50 SPF grade ICR mice,half male and half female,were used.The mice were feed adaptively in the animal room for 1 day,and then were divided randomly.Each group contains one cage for 5 males and the other cage is 5 females.The mice were orally given with a single dose of 20.0g /kg water or ethanol extacts of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata.The mice were observed at 4 hours after treatment everday from 1 day to the end of 15 days.Duringthe observation,the weight of mice and the food intake at 4-7 days and 12-15 days will be measured.15 days after the end of the observation,the mice were killed,and the effects of these above extacts on heart,lung,kidney,liver,spleen,gastrointestinal tract were checked.(3)Long-term toxicity experiment in vivo: 70 SPF male C57BL/6 mice were orally given with ethanol extract of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata(20,40 g/kg)for 60 or 90 days,respectively.Body weight of mcie was recorded weekly for two times.Blood biochemical parameters,liver,kidney,lung and body weight were measured at 60 days and 90 days,respectively.The liver homogenate were used for MDA and MPO detection.Liver haematoxylin & eosin staining was used to evaluate liver injury.Results:(1)Cell viability was determined by MTT in vitro.The results showed that stilbene glucoside,hyperoside,chrysophanol,gallic acid had no effect on cell viability at the concentration of 100?M.Compared with control,polydatin,aloe emodin,physcion and rhaponticin(100?M)decreased cell viability slightly but with no significant difference(P>0.05).Emodin and rhein had obvious inhibitory effect on the cell viability of L-02 cells(P<0.01).Emodin and rhein were cultured by 1 umol,10 umol,25 umol,50 umol and 100 umol concentrations.At 48 and 72 hours,Emodin and rhein decreased cell viability in L-02 cells with a concentration-dependent manner after treated cells for 48 and 72 hours.(2)Acute toxicity experiment in vivoThere was no death in male and female mousemice at 15 days after the mice were treated with water or ethanol extacts of Polygoni multiflori Radixonce.There was no difference in male’s body weight(P>0.05)between 4 days,5 days and 15 days.There were significant difference in famale’s body weight between 7th and15 th day after mice were treated with water or ethanol extacts of Polygoni multiflori Radixonce(P<0.01).The food take in the male mice treated with ethanol extract of Polygoni multiflori Radix was far lower than that in control group.Later,the food take was restored slowly but still lower than in control and other groups.The food take in male mouse treated with water extractions of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata was higher than control mice or mice treated with ethanol extractions of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata.(3)Long-term toxicity experiment in vivo.Animal body weigh and Viscera index: At 60 days and 90 days after treatment,the body weight in mice trated with extrats was decreased significantly(P<0.01 and P<0.05).The liver index was also significantly decreased(P<0.01).At 90 days after treatment,the renal index was significantly decreased(P<0.01).At 60 days after treatment,the lung index in mice treated with exract of Polygoni multiflori Radix Pareparata at high dose was increased significantly(P<0.01)in 60 days.AST and ALT results: At 60 days after treatment,serum ALT activity in mice treated with high dose extracts of Polygoni Multiflori Radix were decreased(P<0.05),serum AST activity in mice treated with extracts of Polygoni Multiflori Radix were no difference(P>0.05).At 90 days after treatment,serum ALT activity in mice treated with low dose of extract of Polygoni multiflori Radix Pareparata were decreased(P<0.01)and the other tend to decreased.serum AST activity in mice treated with extract of Polygoni multiflori Radix were no difference but trend of increase.MPO、MDA analysis:Liver MPO activity and MDA amount were significantly increased as compared with control(P<0.01,P<0.05)at 60 days and 90 days after treatment.TBA amount: Low dose of extrats of Polygoni multiflori Radix Pareparata and high dose of extrats of Polygoni multiflori Radix increased serum TBA amounts at 60 days after treatment(P<0.05).BUN、CREA amounts: Serum BUN and CREA amounts was increased in mice treated with extracts of Polygoni multiflori Radixand and Polygoni multiflori Radix Pareparata at 60 days(P<0.01,P<0.05).And Serum BUN and CREA amounts was decreased in mice treated with extracts of Polygoni multiflori Radixand and Polygoni multiflori Radix Pareparata at 90 days(P<0.01).LDH activity: Low dose of extrats of Polygoni multiflori Radix Pareparata increased serum LDH activity at 60 day after treatment(P<0.01).TP amount:Serum TP amount in mice treated with the extracts of Polygoni multiflori Radix Pareparatawas significantly increased at 60 days after treatment(P<0.05).ALB amount: Serum ALB amount in mice treated with high dose of extracts of Polygoni multiflori Radix and Polygoni multiflori Radix was significantly increased at 60 days after treatment(P<0.05).Hematoxylin & eosin staining: At 60 days,there were inflammatory cell on the liver with high dose of extracts of Polygoni multiflori Radix.A part of cell start karyolysis for the extracts of Polygoni multiflori Radix Pareparata.At 90 d,all the the extracts of Polygoni multiflori Radix treatment had inflammatory cells gathered on the liver.There were some inflammatory cells gradually converged on liver in the partial extracts of Polygoni multiflori Radix.Conclusion: Anthraquinones such as emodin and rhein can induce significant hepatotoxicity,and they may be the main heptotoxic compounds in Polygoni multiflori Radix.Long-term administration of Polygoni multiflori Radix or Polygoni multiflori Radix Pareparata had weak damage on liver.Lung,but such damage will be recoved with the lapse of days.