Stydy On The Effect Of Chinese Traditional Medicine On Lipopolysaccharide Induced Inflammatory Model Based On Metabolic Pathway Of Arachidonic Acid

Arachidonic acid is released by phospholipase A2 hydrolysis membrane phospholipids of unsaturated fatty acids,essential fatty acids.Arachidonic acid has strong biological activity,including prostanoid?prostaglandins?,blood clots alkanes element class?thromboxanes?and leukotriene?leucotrienes?.Class arachidic acid is an important inflammatory substance,involving in the body's immune and inflammatory reaction,plays an important role in the pathophysiology process of many diseases.Inflammation is one of the most common human diseases,in any part of the body and any organization could happen,it is the immune system's first response to infection or external stimuli,can occur in a variety of diseases,including infectious diseases.For anti-inflammatory drugs,arachidonic acid in the network some protein anti-inflammatory chemical drug design has become an important target,but the single target drugs not well control the balance of the network,thus easy to cause side effects.And in anti-inflammatory drugs in the study of Chinese medicine,because of its good efficacy,adverse reactions and rich source of advantage such as less,more and more get people's attention.Confirmed that have a large number of literature,silymarin,ginseng saponin Rd,emodin and curcumin have anti-inflammatory activity,but its anti-inflammatory mechanism from molecular biology research.Lipid omics concept was first put forward for the first time in 2003,as a branch of metabonomics system was studied for the all lipid^[1],fatty acid is an important part of lipid,fatty acid and its metabolites oxide?class arachidic acid?participate in the process of life in a variety of physiological regulation,to participate in the progress of the disease.And set science as a system of research methods,can directly reflect the body biochemical process and the change of state,therefore,from the Angle of omics fatty acid oxidation and its metabolic product research,can directly reflect a more accurate explanation of its role in the disease mechanism,help to the diagnosis and treatment of disease.However,there is no from the perspective of metabonomics to explain the above active ingredient anti-inflammatory effects of Chinese medicine research.This study set up will be based on ultra high performance liquid chromatography?HPLC?-triple level 4 pole mass spectrometry platform,optimization of mass spectrum parameters,establish representative for 59 kinds of arachidonic acid metabolites of quantitative analysis method;Kdo2-induced by Lipid A RAW264.7 cell inflammation model as the research object,the optimization of traditional Chinese medicine?TCM?concentration of active role models,and USES the target class arachidic acid omics technologies,such as study of silymarin,silymarin capsule,ginseng saponin Rd,emodin and curcumin influence lipopolysaccharide induce inflammation model and kind of arachidic acid metabolic changes in cells,expounds the mechanism by which Chinese medicine active ingredients and anti-inflammatory activity.The changes of eicosanoids in RAW264.7 cells with different treatment had been determined within this method based on UPLC-MS and the biological significance had been explained.1.Optimization of triple level 4 pole mass spectrometry conditions and traditional Chinese medicine?TCM?the concentration of the active ingredient intervention lipopolysaccharide induce inflammation model,get 59 kinds of arachidic acid standard MRM ion pairs and DP,CE spectrum parameters,such as,combined with the liquid condition,59 species of arachidic acid in 4.5 minutes to achieve better separation,and has a better peak shape;Find maintain RAW264.7 cell survival rate was 60%⁸0%respectively within the scope of 5 kinds of traditional Chinese medicine active ingredient concentration range.2.Based on establishing the UPLC/MS/MS analysis method and the optimized herbal active ingredient concentration range,we fight phlogistic active ingredient to intervene KLA induced RAW264.7 cell type of arachidic acid metabolites omics analysis.Establish Kdo2-RAW264.7 cells induced by Lipid A inflammation models,with different concentrations of ginseng saponin Rd to intervene,to set up the blank group,inflammation models and ginseng saponin Rd high,medium and low dose group,in the clear liquid medium in RAW264.7 cells class arachidic acid quantitative analysis,through statistical analysis and Kruskal Wallis test is multidimensional Rd ginsenosides in RAW264.7 cells arachidic acid metabolism,the effects of different active ingredients of traditional Chinese medicine for biomarkers for biology.;Select class a total of 5kinds of arachidic acid markers,including PGF2 alpha,5-oxoETE,5-HTET,dhkPGF2a,12-oxoLTB4.Combined with inflammatory signaling pathways,determine the Rd ginsenosides by AA metabolic pathways of COX-2,5-LOX,thus affecting the KLA induced inflammatory cells content of arachidonic acid metabolites.3.Establish Kdo2-RAW264.7 cells induced by Lipid A inflammation models,respectively,with different concentration of silymarin capsule and silymarin intervention,set up the blank group,inflammation models,silymarin capsule?silymarin?,high,medium and low dose group,in the clear liquid medium in RAW264.7 cells class arachidic acid quantitative analysis,through statistical analysis and Kruskal Wallis test is multidimensional silymarin capsule and silymarin on RAW264.7 cell arachidic acid metabolism,the effects of different active ingredients of traditional Chinese medicine for biomarkers for biology.Select silymarin capsule three kind of arachidic acid markers,including PGF2a,TXB2,12-oxoLTB4,Screening of silymarin class one 12-oxoLTB4 arachidic acid markers.Combined with inflammatory signaling pathways,presumably,silymarin can clear oxygen free radicals or through direct effect COX and LOX pathway of peroxidase make oxygen free radical inactivation and exert anti-inflammatory activity.4.Establish Kdo2-RAW264.7 cells induced by Lipid A inflammation models,respectively,with different concentration of emodin and intervention of curcumin,set up the blank group,inflammation model group,high,medium and low dose water drug groups,in the clear liquid medium in RAW264.7 cells class arachidic acid quantitative analysis,through statistical analysis and Kruskal Wallis test is multidimensional TCM active ingredients to drugs in RAW264.7 cells arachidic acid metabolism,the effects of the different active ingredients of traditional Chinese medicine for biomarkers for biology.Extract emodin class 1 kind of 5-HETE arachidic acid markers.Combined with inflammatory signaling pathways,determine its anti-inflammatory effects that are mediated by suppression of 5 LOX activity to reflect.Screening of curcumin class 1kind of PGF2a arachidic acid markers.Combined with inflammatory signaling pathways that curcumin by inhibiting the activity of cox-2 play anti-inflammatory activity.