Preparation Of Pomegranate Punicalagin And Its Subacute Toxicity Experiment And A Preliminary Study On The Mechanism Of Hepatotoxicity

Objective: 1.To study the subacute toxicity of pomegranate peel polyphenol extract,and to evaluate its safety,and to provide the basis for the dose design of long-term toxicity experiment.2.To establish a fingerprint method,to study the different batches of Kashi and Hetian pomegranate herbs.3.Preparation of pomegranate refined by purification process.4.Study on the toxicity of Pomegranate Pear Carboside in three months.In LDLr-/-mice 5.To explore the mechanism of psoriasis pomegranate extract caused by cholestatic hepatotoxicity.Method:1.Pomegranate peel polyphenol extract for 30 days,the general Activity of each group of mice,the monitoring of body weight,food consumption and toxicity,the end of the fasting 24 h to kill 2/3 mice,the remaining mice Observe 2 w after killing.The hematological parameters and biochemical indexes of the mice were observed and the morphological changes of the liver were observed by HE staining.2.The method of HPLC fingerprint was established,and the precision,stability and repeatability were investigated.The similarity analysis of different batches of pomegranate peel in Kashi and Hetian.3.The preparation of pomegranate was carried out and the content of pomegranate was determined by HPLC.4.Psoriasis pomegranate extract on LDLr-/-mice 3 months toxicity test,monitoring the body weight,anatomy of the viscera appearance of the general observation,detection of biochemical indicators,histopathological examination.5.QRT-PCR was used to investigate the mRNA expression of MRP-3,BESP,MRP-2,CYP7A1 and PPARα bile transporters.Results: 1.During subacute toxicity test,the growth activity of each group was normal.The end of the dosing period,each dose group of mice weight weight,food utilization rate compared with the control group,there were no significant differences(P>0.05);the difference of hematological and blood biochemical indexes were no significant specificity.The end of the administration of each dose group and blank control group of mice liver coefficient compared with significant difference(P<0.01),and the recovery of liver coefficient and blank period observed only in high dose group,control group had significant difference(P<0.05),and in the recovery of mice in high dose group liver pathological observation period in the toxic performance,namely hepatocellular spotty necrosis with inflammatory cell reaction,the visible part of the central vein of hepatic lobule and mild dilatation,congestion,liver Kuffer cell proliferation,periportal infiltration ofinflammatory cells.2.Punicagranatum fingerprint method validation results showed that HPLC fingerprint of punicagranatum precision,stability and repeatability are in line with the requirements of the 12 batch of punicagranatum high similarity,different punicagranatum is basically the same in the chemical composition,but there are some differences in the amount of components.3.Kashi punicagranatum contained punicalagin component is higher than the Hotan pomegranate,pomegranate glycosides content at Kashi pomegranate prepared by the same process of leather were more than 80%.4.Pomegranate pomegranate glycosides picoamps extracts administered after 3 months of mouse liver HFD+PU=500mg/kg·d HFD+PU=750mg/kg·d dose group,dose group histopathologic examination showed cholestatic liver injury.5.QRT-PCR in the experiment of mRNA expression,BESP showed down-regulation of mRNA expression,MRP2 showed down-regulation of mRNA expression,MRP3 showed up-regulated expression of CPY7A1 mRNA downregulated expression of PPAR alpha,mRNA showed up.Conclusions:Pomegranate peel polyphenol extract showed liver toxicity in subacute toxicity test.Kashi and Hotan each batch of pomegranate peel similar.Kashi pomegranate pomegranate Picoamp glycosides content in 80%.At higher content of Hotan pomegranate glycosides pomegranate herbs.The pomegranate pomegranate Picoamp glycosides intervention LDLr-/-mice 3 months in liver biopsy in high dose group HFD+PU=500mg/kg·d group and HFD+PU=750mg/kg d group showed a cholestatic liver injury.The pomegranate pomegranate glycosides picoamps mechanism of cholestasis liver injury of MRP3 due to transport substrate and MRP2 has extensive homology,so that the high expression of MRP3 is a kind of compensatory to weaken the function of MRP2.To observe the expression of gene chip suggests further screening of the pomegranate polyphenols and toxicity of liver specific genes related to mRNA,but also can be used to detect protein expression of liver tissue Western blot,intends to fully explore the mechanism for the liver injury induced by cholestasis type from gene level and protein level.