Studies On The Chemical Constituents And Its In Vitro Anti-influenza Virus And Anti-inflammatory Activity Of Mussaenda Pubescens

Background and ObjectiveTraditional Chinese medicine Yuye Jinhua,also known as mountain licorice,for the Rubiaceae(Rubiaceae)jade leaf flower(Mussaenda L.),jade leaf flower(Mussaenda pubescens Ait.f.)dry stem and leaf.According to the study,it is shown that Mussaenda pubescens Ait.f contains chemical constituents such as stigmasterol,?-sitosterol,triterpene acid and Mussaendoside A,which are cold,sweet and light,and go to bladder,lung and large intestine.Solution medicine,dampness medicine.With heat and dampness,solution table swelling,refreshing refreshing,blood circulation and other effects,can be used to treat colds,fever,hot and humid urine,snake bites,treatment of gastrointestinal diseases and prevention of influenza.Yulian Jinhua in recent years due to anti-inflammatory and anti-influenza effect was significantly studied,the experiment confirmed that the water extract of Euonymus japonicus has anti-inflammatory and anti-virus effect,which contains the ingredients of mussaenoside(mussaenoside)Iridoid glycosides,with significant anti-inflammatory,analgesic and antibacterial effects.The experimental results showed that Yuhua Jinhua Decoction could down-regulate the levels of IL-6 and TNF-? in the serum of mice.Influenza virus(IV)belongs to the genus Pseudomonas(Influomyxoviridae)influenza virus genus(Influenzavirus),influenza virus according to the capsule on the hemagglutinin(HA)and neuraminidase(NA)structure and gene characteristics,Can be divided into 16 HA subtypes and 9 NA subtypes.Influenza virus is a pleomorphic capsular virus,spherical,virion structure can be divided into capsule,protein and nuclear shell three layers,outer capsule by a containing three transmembrane proteins(HA,NA,M2)of the lipid bilayer.Among them,HA is the most abundant envelope protein,accounting for 80%,virus adsorption and membrane fusion play an important role.When the influenza virus invades the body,the body in the production of inflammatory cytokines to eliminate pathogenic microorganisms at the same time,in order to prevent excessive inflammation,the body will produce endogenous anti-inflammatory cytokines.Proinflammatory cytokines mainly include IL-1 ?,IL-2,IL-6,IL-8,IL-17,TNF-? and so on.Based on the above research,this study mainly studies the chemical constituents of the golden flower and discusses the anti-inflammatory effect of anti-influenza virus in vitro.MethodsThe compounds were isolated and purified by silica gel,sephadex LH-20 and ODS column chromatography.The structures of the compounds were identified by nuclear magnetic resonance(NMR)and literature comparison.Five compounds were isolated and identified from the three compounds.(P<0.05).The antiviral activity of MDCK was detected by CPE and MTT method.The relative level of HA gene was detected by qPCR method.To evaluate whether the compound has a regulatory effect on the expression of HA gene.In the inflammatory model of macrophage THP-1 induced by lipopolysaccharide(LPS),anti-inflammatory activity was evaluated.In this study,CCK-8 method was used to determine the activity of THP-1 cells and the TC50 of each monomer compound.Real-time-PCR method was used to detect the expression of TNF-8,IL-23.IL-22,IL-1 ? mRNA expression levels.Resultcaffeic acid methyl ester(1),stigmasterol(2),mussaendoside R(3),mussaendoside Q(4),mussaendoside G(5),mussaendoside U(3),mussaendoside(6).The anti-inflammatory activity of compounds 1,3,4,5 and 6 in LPS-induced macrophage THP-1 was selected.The results showed that IL-1 ?,IL-23 and TNF-A,IL-12,IL-22,IL-8 mRNA in the relative content of a certain ability to adjust,in which the regulation of compounds 1 and 3 significant.Compounds 1,3 and 4 have antiviral activity in vitro and have a down-regulation effect on the relative content of HA mRNA.ConclusionCompound 1 was first found from this plant.Compounds 1,3 and 4 have anti-influenza virus activity in vitro and have a mechanism that correlates the relative expression of HA mRNA.In the inflammatory cell model of THP-1 in vitro induced by LPS,it was proved that the monocyte compound of Yuling had a modulating effect on influenza-related inflammatory cytokines and had anti-inflammatory activity in vitro.