Study On Screening, Disassembling And HPLC Analysis Of Radix Hedysari Prescription On Prevention And Treatment Of Liver Injury

Radix Hedysari,also known as solitary root,is the dry root of Hedysarum Polybotrys Hand.-Mazz.,a leguminous plant.It is commonly considered as one famous-region drug of Gansu province in China.After years of research,we have found that Radix Hedysari has various biological activities such as improving immunity,resisting tumor,resisting oxidation,resisting liver fibrosis,and resisting tumor.In addition,the experimental study in the early stage of our laboratory indicated that Radix Hedysari has a remarkable therapeutic effect on CCl₄-induced liver injury in mice.At present,at home and abroad,most of the studies focus on the prevention and treatment of liver injury by monomer compounds of traditional Chinese medicine,single herbs or single traditional Chinese medicine prescriptions.But there are few comparative studies on screening multiple traditional Chinese medicine prescriptions simultaneously,especially it is still a blank in the research on screening and disassembling of multiple traditional Chinese medicine prescriptions with Radix Hedysari as the main herb.The specific contents of this paper are as follows:The first part:screening the traditional Chinese medicine prescriptions of Radix Hedysari for prevention and treatment of liver injury.Liver injury was induced by subcutaneous injection of carbon tetrachloride（CCl₄）in mice.At the same time,4 of the traditional Chinese medicine prescriptions of Radix Hedysari(20 g·kg^-1,once each day)were given intragastrically.After 7weeks,serum alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,liver superoxide dismutase（SOD）and malondialdehyde（MDA）were detected.Histological examination was performed using hematoxylin-eosin staining and light microscopy.The results showed that the effect of CCl₄ on liver injury in mice was obvious,and the effects of the 4 of the traditional Chinese medicine prescriptions of Radix Hedysari on liver injury induced by CCl₄ were significant,but there were differences in their pharmacodynamic activities,among which the traditional Chinese medicine prescription-three of Radix Hedysari was the best and has the value of further study.The second part:the former screened traditional Chinese prescription of Radix Hedysari was disassembled by the method of withdrawing the herb.Starting from the relationship of jun,chen,zuo and shi,taking Radix Hedysari（jun herb）as the core,remove each of the other four herbs as four groups of disassembled prescriptions.Liver injury was induced by subcutaneous injection of carbon tetrachloride（CCl₄）in mice.At the same time,the complete traditional Chinese medicine prescription of Radix Hedysari and four disassembled prescriptions(20 g·kg^-1,once each day)were given intragastrically.After 7 weeks,serum alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,the Ratio of albumin to globulin（A/G）,liver superoxide dismutase（SOD）and malondialdehyde（MDA）were detected.Histological examination was performed using hematoxylin-eosin staining and light microscopy.The results showed that the effect of CCl₄ on liver injury in mice was obvious,and the effects of the complete traditional Chinese medicine prescription of Radix Hedysari and four disassembled prescriptions on liver injury induced by CCl₄ were significant,but there were differences in their pharmacodynamic activities,among which the disassembling prescription-one was the best and has the value of further study.The third part:a HPLC-DAD method was established for the qualitative analysis of the complete traditional Chinese medicine prescription of Radix Hedysari and four disassembled prescriptions.The main chromatographic peaks of the complete traditional Chinese medicine prescription of Radix Hedysari and four disassembled prescriptions were assigned and the composition differences of HPLC spectra were compared.The chromatographic column was diamonsil C18 column（250 mm×4.6 mm,5μm）and the mobile phase was acetonitrile-0.2%acetic acid aqueous solution with gradient elution.The detection wavelength was 330 nm and the column temperature was 30℃.The results showed that HPLC-DAD technique could be used to qualitatively identify and compare the components of the complete traditional Chinese medicine prescription of Radix Hedysari and four disassembled prescriptions by peak ascription.