Research On The Anti-inflammatory Effects And Preliminary Mechanism Of The Yinhua Miyanling Tablet

Yinhuanmiyanling Tablets is one of proprietary Chinese medicinewhich composed by honeysuckle, Scutellaria barbata, knotgrass and otheringredients, is the clinical commonly used drugs for the treatment ofurinary system infections, it has become one of the preferred Chinesemedicine for the treatment of urinary tract infections. Previous studiessuggest that the medicine has the functions of anti-bacteria, detoxification,leaching diuresis, kidney foundation, and blood circulation, improveimmunity and other effects. Our group used modern pharmacologyresearch techniques found that Yinhuamiyanling Tablets of95%ethanolextract inhibits the growth of E. coli, MBC50-90of YinhuamiyanlingTablets of95%ethanol extract on E. coli standard strain ATCC25922andclinical E.coli producing ESBLs were100mg/mL and150-300mg/mL.But whether they have anti-inflammatory effects and theiranti-inflammatory mechanism is unclear. This study from the level ofcytology proved Yinhuamiyanling Tablets have anti-inflammatory effect,and applied the inflammation factor chip preliminarily discussed itsanti-inflammatory mechanism, in order to further promote the drug’sclinical applications and provide experimental and theoretical basis.Methods：1. Anti-inflammatory activity of drugs：By Ficoll densitygradient centrifugation method to separate the peripheral bloodmononuclear cell (PBMC) from healthy people. Adopting Steriledlipopolysaccharide (LPS) stimulates PBMC to establish the inflammation model. Experiment is divided into normal control group (given sterile cellculture medium), model group (LPS inflammatory model group), drugtreatment group (drug treatment after modeling). The normal controlgroup as control，used Tetramethyl azo thiazole blue salt (MTT method)determine drug toxicity in drug treatment group. To confirm the numberof cells before and after drug treatment did not change significantly,weused Trypan blue method counting the rate of living cells，then we usedELISA method to detect the inflammatory cytokines TNF-α，IL-1β，IL-6，IL-10，MCP-1the level of protein expression in each cell cultures.2.Anti-inflammatory mechanism of drugs： By PAHS-097ZA-2RT2ProfilerTMPCR Array Human inflammasomes factor chip method tohigh-throughput screening inflammation factor of drug targets and therelated pathways, application of MatLab clustering analysis method andKEGG database for analysis of screening results.Results: The IC50of Yinhuamiyanling Tablets to PBMC was4.05mg/Ml; the survival rates of cell in each group before and after drugtreatment is more than95%. Drug treatment group compared with modelgroup, the expression levels was significantly reduced of each cytokineTNF-α, IL-1β, IL-6, IL-10, MCP-1(P <0.01). By human inflammasomesPCR Array chip showed that, drug treatment group compared with modelgroup there are15differentially expressed genes appear2.0times moreopposite effect. Pathway analysis showed that these genes were involvedin NOD-like receptor signaling and Toll-like receptor signaling pathway.Conclusion: Yinhuamiyanling by reducing the secretion of IL-1β,and IL-18and other inflammatory factors exert anti-inflammatory effects,its mechanism is the drug affects the NOD-like receptors and Toll-likereceptor Pathway of inflammation-related gene NLRP3, NLRP1expression and caspase-1, etc., thereby inhibiting downstream of the inflammatory response.