| The liver is an important organ of the human body,with producing hormones,tra nsforming endogenous and exogenous substances,detoxification and other functions.W ith the aggravation of environmental pollution,a great deal of harmful substances are harmful to the health of the liver.The toxic effects of exogenous hepatotoxicity on th e liver are manifested with the ability to produce large amounts of free radicals which lead to hepatic lipid peroxidation and necrosis of hepatocytes.Le Sai Pu?LSP?granules consist of chrysanthemum extract,grape seed extract a nd Lycium barbarum polysaccharide extract and other components.Chrysanthemum is t he dry head of Chrysanthemum morifoLium Ramat.Chrysanthemum has a long history in chinese for long time with two applications in eating and medicinal.Chrysanthem um Contains flavonoids,triterpenes and other chemical compositions,which listed as t he top grade in the"Shen Nong's Herbal Classic"with no side effects for long time ta king.Studies have shown that chrysanthemum flavonoids and other substances have a strong antioxidant capacity.Lycium barbarum is the dry and ripe fruit of Lycium barb arum L.,which contains Lycium barbarum polysaccharides and other components.Lyci um barbarum polysaccharide has the effect of preventing liver injury in experimental a nimals.Grape seed is the seed of Vitis viniferaL,and the main component in Grape see d is proanthocyanidins.In the proanthocyanidins,Oligomeric Proanthocyanidins?OPC?h as a strong antioxidant activity.In this study,the quality standard of granules was established and the protective effect on the acute liver injury from carbon tetrachloride and alcohol was studied.1.Using the orthogonal design method to optimize the preparation technology of the particles and establish the quality standard.The results showed that when the ma in drug was 3.6 g,the dosage of dextrin was 5.4 g,the volume of ethanol was 2.0mL and the concentration of ethanol was 90%,the yield of granule was higher and th e optimum preparation technology was determined.2.Making the quality standards of granules,and the quality assessment of Le Sai Pu granules.The granularity,melting resistance,moisture,filling quantity differences were comply with relevant provisions.The content of procyanidins and total polysaccha rides in the preparation were determined by visible spectrophotometry.The procyanidin s content was 83.78-85.63 mg/g,and the total polysaccharide content was 99.53-105.19 mg/g.3.The acute liver injury model in mice induced with Carbon tetrachloride was e stablished.Eighty male mice were divided into 8 groups:normal group,model group,positive group,Le Sai Pu low dose group and high dose group,chrysanthemum grou p,wolfberry group and grape seed group.Normal group and model group were given0.5%sodium carboxymethyl cellulose,the positive group was given Bifendatatum 200mg/kg,the low dose of Le Sai Pu 600 mg/kg,the high dose group of Le Sai Pu 1200 mg/kg,And 400 mg/kg chrysanthemum extract were given to the chrysanthemum gr oup,400 mg/kg lycium barbarum polysaccharide extract was given to the wolfberry gr oup,400 mg/kg grape seed extract was given to the grape seed group,Continuous adm inistration of dosing for 9 d.On the 7th day after administration,the mice in the oth er groups were subcutaneously injected with 5%CCl4?diluted with olive oil?0.1 mL/10 g except normal group,and continued to be administered on the 8th day.After 12h of fasting,the mice were sacrificed,before the 9th days of dissection continue to a dminister once.Blood was taken at 4?for 4 h,then centrifuged at 4000 r/min for 15min,and serum was collected.AST and ALT levels was determined;Dissected the liv er,washing the liver with cold saline,weighing the liver weight,calculate the liver index.The right lobe of the liver was fixed in 4%formalin for morphological analys is.Remaining liver tissues stored at-80°C for homogenization.The contents of SOD,MDA,GSH,GSH-Px and TG in liver homogenate were determined.The results sho wed that the levels of liver index,AST and ALT were decreased?P<0.01 or P<0.05?in he positive drug group,the low dose group and the high dose group of LSP comp ared with model group.SOD,GSH and GSH-Px were significantly decreased?P<0.01?in the model group compare with normal group.MDA and TG were significantly inc reased?P<0.01?compared with normal group.SOD,GSH and GSH-Px were significan tly increased?P<0.01 or P<0.05?in the positive drug group and the high dose group of LSP compared with model group.MDA and TG contents were significantly decreas ed?P<0.01 or P<0.05?in the positive drug group and the high dose group compared with model group.MDA were significantly decreased?P<0.01?in the low dose group compared with model group.GSH was significantly increase in grape seed group co mpare with model group?P<0.01?.Positive group,low dose group and high dose gro up were able to improve the histopathological status of liver tissue in different degree s.4.Eighty male mice were divided into 8 groups:normal group,model group,po sitive group,LSP low dose group and high dose group,chrysanthemum group,wolfb erry group and grape seed group.Normal group and model group were given 0.5%so dium carboxymethyl cellulose,positive group was given Bifendatatum 200 mg/kg,the low dose of LSP given 600 mg/kg main drug,In the high dose group of LSP,1200mg/kg of the main drug was given.The chrysanthemum group give 400 mg/kg of c hrysanthemum extract.The wolfberry group was given 400 mg/kg wolfberry polysacc haride extract.The grape seed group was given 400 mg/kg grape seed extract,intrag astric administration for 7 days.After 1 h after the last administration,12 mL/kg of50%ethanol was administered to the mice except the normal group.After fasting for12 h,the mice were sacrificed and the blood was collected at 4?for 4 h,then centri fuged at 4000 r/min for 15 min.AST and ALT in serum were measured.The liver we re rinsed in cold saline,Weigh the liver weight,calculate the liver index.The right lobe of the liver was fixed in 4%formalin for morphological analysis.The remaining live r tissue was placed at-80°C for homogenization.The contents of SOD,MDA,GSH,GSH-Px and TG in liver homogenate were determined.There is no significant differ ence of liver index in other group compared with the normal group.SOD,GSH and GSH-Px in the model group were significantly decrease compare with normal group ?P<0.01?.AST,ALT,MDA and TG were significantly increase Compared with the nor mal group?P<0.01?.AST,ALT,MDA and TG were significantly?P<0.01 or P<0.05?decrease in positive group and LSP high dosage group compared with model group.SOD,GSH,GSH-Px were significantly increase?P<0.01 or P<0.05?in positive grou p and LSP high dosage group compared with model group.Positive group,low dose group and high dose group were able to improve the pathological status of liver tissu e in different degrees.After the optimization of Le Sai Pu granules preparing technology,the yield and the appearance shape are well,according with the relevant regulations,and the conte nt determination method is reliable.Le Sai Pu has a significant protective effect on al cohol and carbon tetrachloride-induced acute liver injury in mice,and its mechanism i s related to scavenging free radicals,inhibiting lipid peroxidation,stabilizing hepatocyt e morphology and stabilizing cell membrane structure. |
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