Effect Of Flavonoids Extracted From Aurantii Fructus Immaturus On Allergic Asthma

Food allergy is not only an abnormal reaction of the immune system to food,but also a potentially fatal immune response.With the increase of number of people in food allergy,increasingly serious health and economic problems have been taken to individuals,families,and society.Therefore,food allergy has gradually become a public health problem that has attracted considerable attention.How to prevent and treat food allergy has been focused by lots of researchers.The flavonoids of Aurantii Fructus Immaturus(AFI)are a natural source of flavonoids,which could inhibit the formation of thrombus,the spastic contraction caused by histamine,the liver toxicity caused by lipopolysaccharide,and have antioxidant,anti-cancer,anti-tumor and other bioactivities.So far,there is little study on the anti-allergic activity of total flavonoids from AFI.In this work,the flavonoids were extracted from AFI,and the anti-allergic activity mechanism of the flavonoids was discussed through vivo and vitro experiments combined with network pharmacology.Firstly,RBL-2H3 cell degranulation model and OVA induced allergic asthma model in mice were used to explore the anti-allergic activity of flavonoids;Then,the network pharmacological analysis was used to predict the target and possible mechanism of hesperidin,neohesperidin,naringin and narirutin on allergic asthma.The results provided a scientific basis for the application of total flavonoids of AFI in the prevention food allergy.The details were as follows: In vitro,the effect of flavonoids extract from AFI on the morphology of RBL-2H3 cells was investigated by toluidine blue staining;The release levels of histamine and IL-4 of RBL-2H3 cells were measured,and the intracellular calcium influx was observed by fluorescence microscope.The results showed that the flavonoids from AFI in the concentration range of 0-120 ?g/m L could significantly inhibit ?-hex,histamine,IL-4 release and calcium influx of RBL-2H3 cells with concentration dependently.It is proved that the flavonoids from AFI had inhibitory effect on RBL-2H3 cells degranulation.In vivo,OVA was used to induce the mouse asthma model to explore the effect of the flavonoids of AFI on allergic asthma.The effects of the flavonoids on vital signs of mice were investigated by scoring the weight,body temperature,behavior and feces of mice.The effects of t the flavonoids on antibodies and inflammatory factors in mice were investigated by measuring the levels of antibodies in mouse serum and related inflammatory factors in mouse alveolar lavage fluid.The effect of the flavonoids on immune organs of mice was studied by measuring the immune organ index of mice.The lung tissue of mice was stained with HE and PAS to investigate the effect on infiltration of inflammatory cells in mouse lung tissue.The results showed that the flavonoids could inhibit the symptoms of weight loss and hypothermia caused by allergic asthma,significantly reduce the content of antibodies in serum and the level of inflammatory factors in mouse alveolar lavage fluid,and alleviate the infiltration of inflammatory cells in lung tissue,which proved that total flavonoids of AFI had anti allergic activity.Network pharmacological: Hesperidin,neohesperidin,naringin,and narirutin used as key words to inquire and screen of active ingredients.To ascertain the relevant targets of the four bioactive compounds in the extracts,TCMSP and STRING(http://string-db.org)were adopted.Data on allergic asthma related targets were acquired from the Gene Cards.The targets above the median were submitted to the string database for PPI network analysis.Auto-Dock predicts the activities of binding of proteins to flavonoids.The results showed that the key targets of the interaction of hesperidin,neohesperidin,naringin and narirutin and allergic asthma were TNF,CASP 3,STAT3 and so on.The enrichment analysis showed that it was involved in biological processes such as signal transduction.The main signal pathways involved are TNF,PI3K Akt and MAPK pathways.The results of molecular docking showed that the four flavonoids had effects on CASP 3 and TNF-?.It has good binding activity.It is predicted that the four flavonoids may achieve anti-allergic activity by acting on the targets in TNF,PI3K Akt and MAPK pathway.