The Study Of Injury Mechanism Of Liver In Rats Induced By(Processed) Polygonum Multiflorum

In this study, we investigated how did (processed) Polygonum multiflorum impact with the liver in rats through measuring various of hepatic biomarkers, and gene expression of bile transport protein in liver system had been determined. Meanwhile, BRL-3A cell line was used to screen the components of (processed) Polygonum multiflorum, which have probalble inhibition or effect, in vitro and vivo, respectively, and we tried to find out what is the pattern for this phenomenon.ObjectiveTo investigate the long-term effect on the liver in rats with (processed) Polygonum multiflorum, and find out which kinds of components had effects on the cell line both in vitro and vivo.MethodsWistar rats were divided into different groups randomly,10rats each group, gender equant, and oral administration with dosage of2g/ml and1ml/100g of raw Polygonum multiflorum and processed one respectively once a day, after administration for30days and90days, rats were sacrificed, then blood and liver tissue collection would be done. Various kinds of hepatic biomarkers in serum and liver homogenate were detected, examination and analysis of pathological section were done. Meanwhile, gene expression of Mrp2、Mrp3、 Mrp4、Oatp1、Oatp2、Ntcp and Bsep were investigated. Besides, the inhibition of7components of Polygonum multiflorum(gallic acid、5-HMF、stilbene glycoside、emodin、emodin-8-0-β-D-glucoside、physion and physion-8-0-β-D-glucoside) to cell line BRL-3A were revealed by IC50, and cell apoptosis as well as enzyme of supernatant were determined. In order to collect medicated serum, SD rats were divided into several groups randomly,6rats each group, gender equant, oral administration once a day with dosage of2g/ml and lml/100g of processed Polygonum multiflorum for4days, after5min、15min、30min、45min、60min、90min、1h、1.5h、2h、4h、6h、8h、10h and24h since last administration, blood collection were exactly done, then medicated serum of different timing were prepared. Cell inhibition effect、 apoptosis and enzyme of supernatan were investigated after administration of this serum.ResultsThe result of pathological section revealed that after administration of Polygonum multiflorum for90days, light hepatitis, hyperplasia of endothelium and fibroblasts occur in liver tissue. Hyperbilirubinemia is one of the most important marker for this trial. Meanwhile, Mrp3gene were up-regulated significantlly-by both raw and processed Polygonum multiflorum, but the processed one in a lower level. Gender differences should be noticed because of the high sensibility of Mrp3gene in male rats to the drug. At the same time, according to the result of medicated serum trial,1.5h and6h serum had the most inhibitive effect on the cell,2h and8h serum induced obvious apoptosis compare with the blank serum group, and the major damage to the cell membrane was induced by1h and24h serum.ConelusionsBoth of the raw Polygonum multiflorum and the processed one had long-term effect on rat’s liver, including light hepatitis and cholestasis, and the drug had significant effect on Mrp3transport protein in liver tissue. As far as we know, anthraquinone in the Polygonum multiflorum had the most toxicity to the cell among the7components as we investigated, and different times of medicated serum had showed discrepancy extent of cytotoxicity. In brief, long-term administration of processed Polygonum multiflorum which processed by our standard processing may cause cholestasis with the effective dose, but in a lower level compare with the raw one.