Studies On Spectrum-effect Relationship Of Radix Hedysari And Treatment Of Radix Hedysari Polysaccharide On The Carbon Tetrachloride-induced Hepatic Fibrosis And Bone Loss

Radix Hedysari?HQ?,a traditional Chinese medicine,has been widely used since ancient times.The fingerprints of Radix Hedysari extracts from different origins were measured using the high performance liquid chromatography coupled with diode array detector method,by which we identified 3 to 5 kinds of chemical ingredients through a series of optimization of chromatographic conditions.According to the theory of traditional Chinese Medicine,HQ played important roles in invigorating gas for ascending,benefiting blood for promoting production of fluid,promoting circulation for removing obstruction in collaterals.In modern research,we develop the effect of anti-hepatic fibrosis and improve osteoporosis.In combination with a variety of statistical methods,we determined the basic materials of Radix Hedysari for efficacy,screened the superior provenances of Radix Hedysari and provided a basis for the further development of its market value.Radix Hedysari polysaccharide was of hetero polysaccharide,obtained by water solution and alcohol sedimentation method.It has a good effect on improving the immunity,anti-tumor,anti-virus and so on.Modern research shows that liver fibrosis as the original disease also causes a variety of bone related diseases,but there were few reports regarding the relationship of these two diseases.In this paper,we explored the prevention and treatment of Radix Hedysari polysaccharide on the carbon tetrachloride-induced hepatic fibrosis and bone loss,making a preliminary study of connection between them.This paper is divided into the following three chapters:Chapter one:The spectrum-effect relationship on anti-hepatic fibrosis effects of Radix Hedysari Hepatic fibrosis was induced in mice by administering a subcutaneous injection of carbon tetrachloride?40%dissolved in peanut oil?continuously for 35 days at a interval of 5 days?0.1 ml/10 g?.The efficiency of different extracts of Radix Hedysari were determined by the levels of Alanine aminotransferase,Aspartate transaminase,TotalProtein,Albumin,Albumin/Globulin?ALT,AST,TP,ALB,and A/G?in serum and relative liver weight.Subsequently,the grey relational degree analysis and partial least squares analysis were employed to reveal the correlation between chromatographic fingerprint of Radix Hedysari from10 different origins and its anti-hepatic fibrosis efficacy.Results show that all of the different habitats of Radix Hedysari exhibited the favorable anti-hepatic fibrosis effect.The components of adenosine,calycosin and ononin were identified,the effective substances foundation of which were adenosine and calycosin.Chapter two:Spectrum-effect relationships on PBM-enhancement?peak bone mass?effects of Radix Hedysari PBM is the peak amount of bone acquired by one when he is mature,after maturing stage PBM degreased gradually.PBM is the basic standard of the research on the diagnosis of osteoporosis.Based on this,we chose baby rats in the experiment,gave them Radix Hedysari extracts from their childhood to adulthood when the bone mass reached the maximum.Then the bone indicators of biomechanic such as bone elastic modulus,yield strength,maximum load and bone mineral density were measured.The serum indexes such as blood calcium,phosphorus,alkaline phosphatase,tartrate resistant acid phosphatase?Ca,P,ALP,TRAP?were investigated.All of the indexes were chosen for comprehensively reflecting the change of peak bone mass.Using grey correlation degree method to explore bioactive substances in Radix Hedysari,while using factor analysis and cluster analysis to select superior provenances of Radix Hedysari.The results showed that 10 batches of medicinal materials of Radix Hedysari could all improve the peak bone mass in rats.All of the common compounds were divided into three parts including three sections.The ingredients of ononin,isoflavone,adenosine,campanulin,formononetin were in high section,which had contributed to the effect.Optimal producing area was Guanting town in Longnan city.Chapter three:Prevention and treatment of Radix Hedysari on the carbon tetrachloride-induced hepatic fibrosis and bone loss The mice was caused by a subcutaneous injection of 20%CCl₄ dissolved in olive oil solution,while given Radix Hedysari extract daily.Continuous 35 days later,the indexes of liver fibrosis,bone markers were determined and liver histological of tissue pathology were operated by hematoxylin eosin?HE?staining and masson trichrome staining?MASSON?.The rank sum test of ridit analysis was used for one-way analysis of variance?one-way ANOVA?.The bone indexes in serum were measured for real-time monitoring the change of bone mineral content and the hydroxyproline in femur was detected using digestion method.The results indicated that liver fibrosis in mice could also cause bone loss and was the primary cause of bone loss in this experiment.Radix Hedysari polysaccharide can significantly improve the hepatic fibrosis and bone loss induced by CCl₄ in mice.