Study On Active Components And Mechanism Of The Anti-inflammatory Effective Parts Group From Smilax China

Traditional Chinese medicine Smilax,also known as Smilax China for liliaceous plant Smilax China L.dried roots.As a commonly used traditional Chinese medicine in China,the records in the 2015 edition of "Chinese pharmacopoeia",for the effects of heat-clearing and detoxifying,softening and resolving hard lumps,Hubei Furen Pharmaceutical Co.Ltd.in Smilax China for raw materials for the production of "Jingangteng capsule" for treatment of pelvic inflammatory disease,used in the treatment of pelvic inflammatory disease,adnexitis,bag piece and other disease of department of gynaecology,curative effect is distinct,welcomed by the masses of patients.Project previous studies showed that saponins and flavonoids ingredients are major smilax anti-inflammatory effective material parts,but to play the pharmacodynamic effects of active ingredient is not clear.Based on the research of smilax anti-inflammatory effective parts of fingerprint and serum of medicine research results,this thesis of smilax anti-inflammatory effective parts of main into the blood components of anti-inflammatory,antioxidant and immune regulating activity research,and the main active ingredient of anti-inflammatory mechanism research,the effects of the anti-inflammatory effect of interpretations of smilax material basis and provides a scientific basis for selecting the active lead compounds.This thesis mainly completed the following research work:1.Using DPPH free radical,hydroxyl free radical reaction system,determination of FRAP method respectively smilax effective parts group and astilbin,taxifolin,resveratrol,rutin,quercetin,huang qi glycosides and so on six monomer composition of radical scavenging ability and iron ion reduction ability;Establish H₂O₂ induced rat red cell membrane damage model,after incubation test drugs on erythrocyte hemolysis inhibition rate andmalonaldehyde?MDA?in the red blood cell suspension content,superoxide dismutase?SOD?and glutathione peroxidase?GSH-Px?activity.Results show that the anti-inflammatory effective parts of smilax have antioxidant effect in vitro and its six monomers,and the red cell membrane protective,speculation smilax anti-inflammatory effects related to its antioxidant properties.2.The mice spleen lymphocyte immune activity screening model,explore the effective parts of smilax and contain the taxifolin,resveratrol,methyl raw yam saponins and the effects of 12 monomer compositions on the immunoregulatory,with CCK-8 method was developed for the determination of cell proliferation rate of cells.With Elisa method in IL-2 and TNF-? and IFN-? three secretion of cytokines,the results show that these drugs are immune activity,including engeletin and saponins of class composition activity more prominent,and showed that the synergy of many ingredients of traditional Chinese medicine.3.Construction of COX-1 and COX-2 molecular targets based on THP-1cells in vitro anti-inflammatory screening model,add smilax effective parts group and its effect on the 12 monomer compositions in THP-1 cells.The results showed that these drugs on COX-2 inhibit certain selectivity.Results show that these drugs to the inhibition of COX-2 all have certain selectivity,and smilax effective parts group,taxifolin and astilbin of the most significant inhibition of COX-2.4.With LPS induced THP-1 cells model is adopted to explore the smilax effective parts group,astilbin and taxifolin,the related proinflammatory cytokines generated and the impact of gene expression.Results show that the three kinds of active ingredients can significantly inhibit IL-1?,IL-6 and TNF-? cytokine levels and its mRNA expression.This could fathom,smilax active ingredient is a way to give play to the role of anti-inflammatory inhibition of cells to produce IL-1?,IL-6 and TNF-? proinflammatory factor,etc.5.Using Western blot method to detect smilax effective parts group,taxifolin and astilbin of MAPK signal transduction pathway of P38 lightning,ERK1/2,JNK and the influence of the amount of phosphorylated protein expression.The results show that compared with LPS group,smilax effective parts group,taxifolin and astilbin of phosphorylated can make P38,JNK,ERK1/2 protein expression decreased,leading to cell related inflammatory factor secretion is reduced,achieve anti-inflammatory effect.