Studies On Pharmacodynamics And Active Constituents Analysis In Crude And Processed Cornus Officinalis

Cornus officinalis,derived from the dry ripe sarcocarp of Cornus officinalis Sieb.et Zucc.,is one of the important traditional chinese medicines and used clinically as Crude drug and Processed product which is produced after steaming the crude drug pre-steeped in wine.Crude drug is good at preserving yin and arresting sweating,and applied to spontaneous perspiration and night sweating with yin asthenia.Processed product, enhancing the effect of invigorating kidney and liver,is used commonly to treat vertigo, tinnitus,impotence and seminal emission,uroclepsia,thamuria,metrorrhagia,soreness and weakness of the waist and knees.The main active components contain organic acid,iridoid glycoside and polysaccharides.This paper aims to study Cornus officinalis before and after processing around its intrinsic changes in the chemical composition,looking for the active componenets changing during the processing（contents changing,newly increasing or disappeared）,to explain the difference of the effects of Cornus officinalis before and after processing.This paper is divided into five parts to study and analyse the pharmacodynamics and the effective components of Fructus Corni before and after processing.The first part:the comparison studies on the pharmacodynamics of Cornus officinalis before and after processingWe studied the influences of Cornus officinalis before and after processing on the immune function of immunocompromised mice,the kidney-yin deficiency mice and the protective effect of acute liver injury.The results showed that processed products decoction in high doses significantly increased mice specific humoral immune function enhancement; processed products decoction in high doses could extend kidney-yin deficiency mice weight loading swimming time,hypoxia endurance time,increase serum SOD activity of kidney-yin deficiency mice and reduce the content of MDA;processed products decoction in moderate doses had significant increase to protect acute liver injury.The second part:the active components analysis of Cornus officinalis before and after processing1.The comparison studies on the RP-HPLC-DAD chromatograms of Cornus officinalis before and after processing We analysed and comparised the differences of RP-HPLC-DAD chromatograms of Cornus officinalis before and after processing.After being processed with wine,the contents of gallic acid and 5-hydroxymethylfurfural increased largely;whereas the contents of morroniside,sweroside and loganin decreased;In addition,the content of undetermined compound 1 increased markedly,the content of undetermined compound 2 decreased slightly.2.The identification of undetermined compoundsWe first analysed the two compounds by using LC-MS technique,obtained several possible molecular formula,but it is insufficient to determine the compounds.Then we prepared and isolated the compounds by using high performance preparative liquid chromatography,and finally got one compound（undetermined compound 2）.It was detected by ¹³C-NMR,¹H-NMR and 2D C-H COSY,and identified as cornuside.3.Studies on changes of content of main active compositions of Cornus officinalis before and after processingWe simultaneously determinated six active components in crude and processed Cornus officinalis by RP-HPLC-DAD,and investigated the effects of water decocting and methanol ultraphonic extracting on the contents of the components in Cornus officinalis.The third part:the studies on the pharmacodynamics of cornusideCornuside could promote the proliferation effect of mice mononuclear macrophage after 24h,but the proliferation effect was inhibited after 48 h or 72 h;It could also protect the astrocyte induced by H₂O₂ and mesangial cell induced by glucose.The fourth part:the studies on fingerprint of crude and processed Cornus officinalisBy optimizing of chromatographic conditions,we established the common fingerprint mode of Cornus officinalis before and after processing from 12 batch of crude drug and 12 batch of processed products by HPLC.There were 19 common peaks in crude drug and 5 peaks identified;there were 23 common peaks in processed product and 6 peaks identified amples.The fifth part:the studies on the Serum Pharmacochemistry and Serum Pharmacology of Cornus officinalis before and after processingA chromatographic method was established to analyse the components of Cornus officinalis being administrated into rat in vivo.Through analysing fingerprints of 0～240min serum cotaining medicine after administration,we found that the components of 5-HMF,morroniside and loganin entered into the blood,gallic acid,sweroside and cornuside were not detected.The study of Serum Pharmacology showed that the serum cotaining medicine of Cornus officinalis before and after processing had apparent protection to hepatic cells injury induced by H₂O₂,moreover,the effect of serum cotaining medicine of crude drug was inferior to that of processed product;and also had apparent protection to mesangial cells injury induced by CdCl₂,moreover,the effect of serum cotaining medicine of crude drug was inferior to that of processed product as well.