Protective Effect Of Total Flavonoids Of Penthorum Chinense Pursh On Chemical Liver Injury And Preparation Of Capsule

Objective:To investigate the protective effect of total flavonoids of Penthorum chinense Pursh on chemical liver injury in mice,three chemical liver injury models were established,including acute chemical liver injury caused by carbon tetrachloride,acute drug liver injury caused by acetaminophen and subacute alcoholic liver injury caused by alcohol.And to prepare total flavonoids of Penthorum chinense Pursh capsules for the development of safe and effective anti-chemical liver injury drugs and health food to provide basis.Methods:1.Protective effect of total flavonoids of Penthorum chinense Pursh on chemical liver injury:A total of 170 SPF Kunming mice were selected to establish the chemical liver injury model using carbon tetrachloride（CCl₄）;acetaminophen（APAP）and alcohol respectively,In the CCl₄and APAP-induced injury model experiment,mice were divided into five groups:normal group,model group,positive control bifendate group（150 mg/kg）,high dose and low dose groups of total flavonoids of Penthorum chinense Pursh（800,400mg/kg）.Each group was given intragastric administration once a day.CCl₄group was given continuous administration for 21 days,and APAP group was given continuous administration for 14 days.1 h after the last administration of the corresponding drugs in the two experimental groups,except for the normal group,no modeling intervention was made,other groups were intraperitoneal injection of 1%CCl₄olive oil solution（10 m L/kg）and APAP（300 mg/kg）;In the alcohol-induced injury model,in addition to the above 5 groups,two groups were added:extract of Penthorum chinense Pursh group in high-dose group（3400 mg/kg）and extract of Penthorum chinense Pursh group in low-dose group（1700 mg/kg）.In the morning,the mice were given 50°Chinese Baijiu intragastrically,and in the afternoon,the mice were given according to the prescribed regimen for 16 consecutive days.In all three experiments,after the completion of the last administration,the mice were fasted without water for 16h,and then the mice were sampled and killed.The levels of ALT,AST,LDH,ALP,TG and TC in serum of mice were detected by automatic biochemical analyzer according to the instructions of the kit,and the levels of TNF-α,IL-6,SOD,GSH and MDA in liver tissue were detected by ELISA,HE staining to observe the liver pathology organization change.2.Preparation and quality evaluation of total flavonoids of Penthorum chinense Pursh capsule:The capsule molding process was optimized with Angle of rerest,hygroscopic rate and molding rate as indexes.Capsules of total flavonoids of Penthorum chinense Pursh were prepared and the quality of the capsules was evaluated.3.Protective effect total flavonoids of Penthorum chinense Pursh capsule on acute chemical liver injury induced by carbon tetrachloride in mice:Select 70 SPF kunming mice were divided into normal group,model group,positive control bifendate group（150 mg/kg）and total flavonoids of Penthorum chinense Pursh high dose group and low dose group（400,200 mg/kg）and total flavonoids of Penthorum chinense Pursh capsule high dose group and low dose group（800,400 mg/kg）.Each group was given intragastric administration according to the proposed drug and corresponding dose（once a day）,1 h after the last administration of the corresponding drugs,except for the normal group without modeling intervention,the other 5 groups were ip1%CCl₄olive oil solution（10m L/kg）,16 h later,blood samples were taken and liver weighed,after that,the levels of ALT,AST,LDH and ALP in serum of mice were tested by automatic biochemical analyzer.Results:1.Protective effect of total flavonoids of Penthorum chinense Pursh on chemical liver injury:Chemical liver injury induced by CCl₄and APAP resulted in higher liver coefficient in model group than in normal（p<0.01）,the serum related enzyme indexes ALT,AST,LDH and ALP in model group were significantly higher than those in normal control group（p<0.05,p<0.01）,while the liver tissue related indexes TNF-α,IL-6 and MDA were significantly increased（p<0.01）,in addition,the SOD level was significantly decreased（p<0.01）.Compared with the model group,enzyme indexes ALT,AST,ALP,LDH,TNF-α,IL-6 and MDA in serum were significantly decreased（p<0.05,p<0.01）,in addition,SOD level in liver tissue were significantly increased in high dose group and low dose group of total flavonoids of Penthorum chinense Pursh（p<0.05,p<0.01）,and pathological state of liver tissue was significantly improved.Alcohol induced liver injury in model group increased liver coefficient（p<0.01）,The levels of ALT,AST,LDH,ALP,TG and TC in serum in model group were significantly higher than those in normal group（p<0.01）,The levels of TNF-α,IL-6 and MDA in liver were also significantly higher than those in normal group（p<0.01）,while SOD and GSH liver tissue were significantly lower than those in normal group（p<0.01）;Compared with model group,ALT,AST,ALP,LDH,TG,TC in serum were significantly decreased（p<0.05,p<0.01）,in addition,TNF-α,IL-6 and MDA,SOD and GSH levels in liver tissue were significantly increased in high and low dose group of total flavonoids of Penthorum chinense Pursh（p<0.05,p<0.01）;The pathological status of liver injury in mice was significantly improved.The serum ALT,AST and LDH levels in the total Flavonoids of Penthorum chinense Pursh group were significantly lower than extract of Penthorum chinense Pursh group with the same crude drug（p<0.05,p<0.01）.2.Preparation and quality evaluation of total flavonoids of Penthorum chinense Pursh capsule:The Angle of repose,hygroscopic rate and granulation were used as excipients forα-lactose,and the drug-auxiliary ratio was 1:1.The Angle of repose of anhydrous ethanol with 10%dosage was 28.89°and the heap density was 0.55 g/m L.No.0 capsule was selected for filling,and the critical relative humidity was 81%.The water content（<9%）,the difference of loading amount（±10%）and the disintegration time（<30min）were all in line with the requirements of pharmacopoeia,the average content of total flavonoids in the capsule was 143.36mg/g.3.Protective effect total flavonoids of Penthorum chinense Pursh capsule on acute chemical liver injury induced by carbon tetrachloride in mice:Chemical liver injury induced by CCl₄liver coefficient model group,and the amount of ALT,AST,ALP and LDH of model group was significantly higher than that of normal group（p<0.01）,compared with model group caused by CCl₄,total flavonoids of Penthorum chinense Pursh capsule group high and low dose group and raw material total flavonoids of Penthorum chinense Pursh group high and low dose group ALT,AST and LDH were significantly lower（p<0.01）,and the ALP of total flavonoids of Penthorum chinense Pursh capsule in high dose group and raw material total flavonoids of Penthorum chinense Pursh in high dose group decreased significantly（p<0.05）.There was no significant difference in reducing serum ALT,AST,LDH and ALP between the total flavonoids of Penthorum chinense Pursh capsule and the raw material（p<0.05）.Conclusion:Total flavonoids of Penthorum chinense Pursh has good protection on mice liver injury caused by carbon tetrachloride and acetaminophen,and its mechanism of action may be related to its inhibiting oxidative stress and inflammation;The protective effect on alcoholic liver injury of total Flavonoids of Penthorum chinense Pursh is better than Penthorum chinense Pursh,and the mechanism of action may be associated with its regulation of lipid metabolism,inhibition of the oxidative stress reaction and the inflammatory reaction.The preparation route of total flavonoids of Penthorum chinense Pursh capsule was reasonable and feasible,and it had protective effect on chemical liver injury,and the effect was similar to total Flavonoids of Penthorum chinense Pursh.