The Study On The Extraction Technology Of Paris Polyphylla And Verbena And Protective Effects On Rats With Alcoholic Liver Injury

In the literature search of alcoholic liver disease,it is found that there are a lot of literatures about alcoholic liver disease and related drugs reported at home and abroad,and most of them are unilateral or compound based on traditional chinese medicine.Compared with the drug treatment under the guidance of modern medical theory,traditional chinese medicine has potential advantages and broad prospects for the prevention and treatment of alcoholic liver disease. According to the screening of traditional chinese medicine prescriptions,in this paper,with Verbena and Paris formula,This topic prescribes with the Chinese herbal medicine of verbena and paris.In this paper,the extraction process of the mixture are researched;By the experiment mode of the rats which are exposed to the stimulation of alcohol and the extract at the same time,the preventive effect on alcoholic liver injury by the mixed extract of verbena and Paris are studied,it was found that the extract had a significant preventive effect on liver;Ethanol induced liver injury model was induced by alcohol stimulation in rats,and then the extract are administered by gastric perfusion,to investigate the pharmacological effects of extract on the formation of alcohol induced liver injury and try to explore its mechanism. 1、Experimental method 1.The extraction of drugs(1) The determination method of Verbena total flavonoids and saponins of Paris polyphylla are established by UV spectrophotometry and HPLC method.(2) The Verbena,Paris polyphylla,a mixture of both,which are extracted Respectively,through the established method of determination to study the influence between the two traditional chinese medicines which are mixed extraction.(3) The verbena and Paris polyphylla is mixed equal after crushed,then single factor experiment of ethanol concentration,ratio of liquid to material,extraction time and extraction times were carried out,and the optimal extraction conditions were determined.(4) Based on the single factor experiments,different experiments levels are seted referenced to the optimal conditions.The extraction times were 3 times,three factors(ethanol concentration,solid-liquid ratio,extraction time)three levels of orthogonal test,the best extraction conditions were determined;The orthogonal experiment of three factors(ethanol concentration,solid-liquid ratio and extraction time)was performed,and ultimately determine the best extraction conditions.(5) In the end,the optimal extraction conditions were verified to determine the extraction efficiency. 2.The liver preventive effect under the stimulation of alcoholThe extracts are used as the experimental drug.While rats were exposed to alcohol,the high,medium and low doses of three doses of experimental drugs and the positive control drugs were administered orally 8 consecutive weeks.The growth of rats,liver coefficient,serum ALT,AST,TC,TG and liver tissue SOD,MDA and other indicators were observed,to evaluate the preventive effect of experimental drugs in liver under the stimulation of alcohol. 3.The protective effect of alcoholic liver injuryThe extracts are used as the experimental drug.The alcoholic liver injury model was induced by alcohol stimulation for 8 weeks.The high,medium and low three dosages of experimental drugs and the positive control drug were administered orally for 4 weeks.The growth of rats,liver coefficient,Liver pathological section observation,serum ALT,AST,TC,TG,i NOS,TNF-α,liver tissue SOD,MDA,GSH,CAT and other indicators were observed,to evaluate the protective effect and mechanism of experimental drugs in treatment of the liver injury induced by alcohol. 2、Experimental results 1.The extraction of drugs(1) During the process of mixing extraction,the extraction process of two kinds of herbs are relatively independent,has no obvious effect on the other.(2) The best extracting process of various single factor extraction were as follows:80% ethanol concentration,30 times of the proportion of the liquid,extraction time 2h,extraction times of 3 times.(3) The optimum extraction conditions of orthogonal test of extracting drugs were as follows:ethanol concentration 70%,solid-liquid ratio of 25 times,extraction time 2.5h,extraction times of 3 times.(4) Extraction results of the verification test shows that average extraction rate of Verbena total flavonoids was 7.054%,RSD =0.008;While the average extraction rate of Polyphyllin I was 0.646%,RSD=0.001.The total extraction rate was 7.670%,which was higher than any single factor experiment. 2.The liver preventive effect under the stimulation of alcoholExperimental drugs can improve the growth of rats which are stimulated by alcohol,reduce the liver coefficient,significantly reduced levels of ALT,AST and TG in serum,significantly decreased the level of MDA in liver tissue,and significantly increased the level of SOD,aren’t observed had a significant effect on level of TC in serum. 3.The protective effect of alcoholic liver injuryExperimental drugs can improve the growth of rats which are stimulated by alcohol,reduce the liver coefficient,significantly reduced serum ALT,AST,TG,i NOS and TNF-a level;significantly decreased the level of MDA in liver tissue,significantly increase the level of SOD,GSH,CAT.Liver pathological section HE staining results showed that experimental drugs can significantly improve the liver cell ultrastructure,decreasing liver cell necrosis,edema and steatosis. 3、Conclusion(1) The simultaneous extraction of total flavonoids from Verbena and polyphyllin with ethanol way is feasible,which with the promotion of significance.(2) The experimental drug can against the lots of free radical produced by ethanol metabolic processes,reduce oxidative stress in liver,protect the liver membrane system,reducing liver lipid peroxidation,reducing the exudation of cell injury and cell contents,so as to decrease the levels of ALT and AST.(3) The experimental drug can eliminate free radicals,reducing oxidative stress in liver,decrease the levels of i NOS and prevent the formation of stronger free radical,thereby reducing the free radical scavenger SOD,anti lipid peroxidation GSH,the consumption of antioxidant enzymes CAT content.(4) The experimental drug can reduce free radicals,raised the level of antioxidant,reduce the liver cell membrane lipid peroxidation which caused by the alcohol metabolism process,thereby reducing membrane peroxidation end product MDA level.(5) The experimental drug can reduce the high oxidative stress status of membrane lipid peroxidation to stability of membrane system,reduce the through of endotoxin and lipopolysaccharide via the hepatic portal vein system into the liver,thereby reducing the activation of Kupffer cells and macrophages,down-regulation of inflammatory mediators TNF-αlevels,reduce the direct damage from inflammation of liver cells.(6) The experimental drug can scavenging free radicals,reducing oxidative stress status,increase the level of antioxidant and reduce membrane lipid peroxidation,downregulation the level of immune mediators etc,correcting and treating liver cells,to restore normal physiological state,maintain the normal physiological function,enhanced compensatory mechanism,improve metabolism,metabolism of sugar,fat,protein and nucleic acid material which are affected by alcohol metabolism,can also essential to maintain normal metabolism,thus TG levels also declined.