The Study On Citrus Aurantium Flavonoids Inhibitory Effect On Lps-induced Inflammatory Mediators In Microglia And Itsunderlying Me·Chanism

Citrus aurantium Rutaceae lime from Citrus aurantium L.and its cultivars or Citrus sinensis Osbeckr dried unripe fruit.With slightly cold,bitter,acrid,sour properties.It has efficacies of dispersing chest qi to relieve chest tightness,eliminating food product to relieve fullness.It was firstly recorded in the "Shen Nong's Herbal Classic" as one of Liu Chen.Citrus aurantium contains flavonoids,alkaloids,volatile oil,polysaccharides..Flavonoids content has higher concentration,which were considered to play an important pharmacodynamic effects material basis in previous study,such as anti-inflammatory,anti-fungal,anti-viral,anti-tumor.Therefore we choose Citrus aurantium flavonoids as the research object,and study its anti-inflammatory effect and mechanism of inhibition of LPS-induced microglial,contents as followed:1.Extraction and separation of flavonoids in 95%Ethanol was used to reflux Citrus aurantium powder.Petroleum ether and n-butanol extracts were separated by column chromatography using silica gel.polyamide,LH-20 column chromatography and column chromatography,to give 7 species known compounds,namely Citrus factors.Hesperetin,Nobiletin,Naringerin,Tangeretin,Hesperidin,Neohesperidin,and Naringin new.2.activity screening and structure-activity relationships of flavonoids on LPS-induced microglia analysisMTT assay was carried out to screen flavonoids inhibition of LPS-induced microglial inflammatory activity;primary microglia expressing cytokines by enzyme-linked immunosorbent assay(ELISA)measurement show that:among seven kinds of flavonoids,aglycone have stronger activity than glycoside,and red-orange pigment and nobiletin with strong activity,both structures were 4'.5,6,7,8-pentamethyl flavonoid 3',4',5,6,7,8-hexamethoxy flavonoids,only nobiletin in the C ring more than a 3' methoxy substituent.3.Study of Tangeretin anti-neurogenic inflammation mechanism Red-orange pigment was the research object.We established LPS-induced microglial cells and primary small glioma cell lines BV-2 model and applicated Griess method,ELISA,RT-PCR and Western blot methocd to explore the recd-orange pigment anti-neurogenic inflammatory mechanism.The results showecd that:the red-orange pigment was in a dose-dependent reduction in LPS-induced microglia iNOS.COX-2 protein expression and NO.PGE2 release;it has dose-dependent inhibition of TNF-?,IL-1?,and IL-6 gene expression;it can inhibit the MAPK phosphorylation levels including three proteins ERK1?JNK and p38 in MAPK;it can inhibit NF-?B nuclear translocation.This paper study suggested that in flavonoid-rich Citrus aurantium the red-orange pigment was more important than other flavonoid aglycone.Red-orange pigment inhibited LPS-stimulated activation of microglia secrete inflammatory cytokines TNF-?,IL-1? and IL-6 and NO,and by inhibiting NF-?B and MAPKs two signaling pathways play a role in anti-neurogenic inflammation which provides a strong scientific experimental basis,to further study the anti-inflammatory activity of flavonoids in Citrus aurantium and its mechanism of action...