Study On Chemical Constituents And Anti-inflammatory Activity Of Viola Yedoensis Makino

Viola yedoensis is a perennial herb belonging to Viola L.genus in Violaceae family.Being one of the traditional Chinese medicines,the dry whole grass of Viola yedoensis can be used as medicine,which has the effects of antisepsis and anti-inflammation,clearing away heat and toxic materials,cooling blood and detumescence.Modern pharmacological studies have shown that Viola fruticosa and some of its active ingredientts have such good activities as antiviral,anti-inflammatory,antibacterial and antitumor,etc.The current research on the chemical constituents and pharmacodynamics of Viola yedoensis are limited and its effective part is still unclear.Therefore,this study conducted the research from the following three parts to study its chemical constituents and anti-inflammatory activity:separation and identification of the components,in vivo anti-inflammatory activity study and its effect on the anti-inflammatory activity of lipopolysaccharide（LPS）-induced in vitro inflammation model of host cells.Objective:To understand the medicative chemical composition of Viola yedoensis thus to study its anti-inflammatory activity.To screen the anti-inflammatory active part and to explore the anti-inflammatory effects in vitro and in vivo of the ethyl acetate part.Methods:（1）The extraction method using solvents of different polar and ethanol cyclotron condensation method were adopted to extract the chemical constituents of the whole grass of Viola yedoensis.The test tube method and the circular filter paper method were used for qualitative pretest analysis of the solvents extracted by the above two different methods,thus to determine the chemical composition of Viola yedoensis.（2）The solvents were concentrated under reduced pressure to prepare the extractum,which was then weighed and the extraction rate was calculated.The extracts from different parts of Viola yedoensis extracted by the cyclotron condensation method were given to the mice.Then such animal experiments were conducted to study the anti-inflammatory effects of the extracts from different parts as pinna swelling induced by xylene,permeability change of abdominal capillary induced by 1%glacial acetic acid,acute inflammation modeling performed by carrageenan-induced rat toe edema,content detection of inflammatory tissue PGE₂,content detection of NO,etc..（3）The extract of ethyl acetate part in Viola yedoensis extracted by ethanol cyclotron condensation method was used to intervene the in vitro inflammatory model of lipopolysaccharide（LPS）-induced RAW264.7 cell.Then its effects on the release of NO,tumor necrosis factor（TNF-α）,IL-1β,IL-6,TNF-α,iNOS and COX-2expression on RAW264.7 cells were investigated,thus to analyze the in vitro anti-inflammatory activity of different solvent extracts of Viola yedoensis.Results:（1）It was detected that the Viola yedoensis solution contains such ingredients as phenols and tannins,alkaloids,organic acids,oil or fat,coumarins and lactones,flavorrnoids,saponins,etc.It was detected that the extract of ethyl acetate part extracted by ethanol cyclotron condensation method mainly contains organic acid,flavonoids and other compound components and the extraction rate in ethanol cyclotron condensation was higher than that in the extraction method.（2）The effect of inhibiting pinna swelling induced by xylene using appropriate concentration of solution extracted from ethyl acetate part of Viola yedoensis through cyclotron condensation method was significantly different from that in the normal saline（NS）group（P<0.05）.In the whole process of monitoring the inhibition rate of toe edema in mice,the difference between the ethyl acetate group and the NS group was extremely significant（P<0.01）.There were also tendencies of inhibiting toe edema in both the petroleum ether extraction part group and the aqueous phase group,but the difference was not obvious.In the capillary permeability test induced by intraperitoneal injection of 1%glacial acetic acid in mice,the optical density（OD value）of the ethyl acetate group was significantly different from that in NS group（P<0.05）and the difference was the most significant.（3）In the experiment of detecting NO content in inflammatory tissues of mice,compared with the control group,the ethyl acetate part significantly reduced NO released into inflammatory tissues（P<0.05）.Conclusion:The chemical constituents of Viola yedoensis were found to be composed of phenols and tannins,alkaloids,organic acids,oils,coumarins and lactones,flavonoids and saponins thus to study its anti-inflammatory activity.Preliminary study showed that ethyl acetate fraction extracted by cyclotron condensation could downregululate the expression of inflammatory factors such as IL-1beta,IL-6 and TNF-a,and down-regulate the expression of TNF-alpha,COX-2and iNOS,which may be one of the reasons for the anti-inflammatory effect of Viola yedoensis.