Network Pharmacology-based Study On The Material Basis From The Herbs Of Mahuang-Fuzi-Xixin Decoction On Treating Allergic Rhinitis

BACKGROUDAllergic rhinitis(AR),is nasal mucosa of allergic diseases,and can cause a variety of complications.It is a variant of the susceptible individuals exposed to the allergen or allergens,known as chronic inflammation of the nasal mucosa.Clinically,nasal congestion,nasal itching,watery nose,sneezing are the main features.its incidence relates to the disorders of body-specific mechanism.Guangdong is the high incidence of allergic rhinitis;the incidence rate is about 15%.Studies have shown that the incidence of global AR is gradually increased in the past 30 years.AR can indirectly increase the incidence of sinusitis,otitis media,and bronchial asthma.Therefore,it attracts widespread attention.In western medicine,desensitization therapy,nonspecific therapy,nerve block therapy is the main methods to treat allergic rhinitis.Non-specific treatment mainly used antihistamines,sugar corticosteroids,decongestants,mast cell stabilizers,and some others,like leukotriene inhibitors,cytokine modulators and antagonists of inflammatory mediators.Currently,glucocorticoid steroids and antihistamines are the main drugs in the treatment of AR.These drugs can only control the symptoms of AR,cannot achieve radical purpose and have obvious side effects.The nasal corticosteroids can cause nasal dryness,local side effects of nasal septum perforation,bleeding and worse,can cause systemic side effects,such as the hypothalamus-pituitary-adrenal axis suppression,inhibiting the growth of children,cause cataracts and glaucoma and other eye complications.Side effects of antihistamines is more obvious,such as the patient can cause drowsiness,headache,dizziness,decreased ability to sense and manipulate and cause side effects in the cardiovascular system.Traditional Chinese Medicine treatment,with respect to Western medicine,is safe,reliable,little side effect,lasting effect.Therefore,it is believed by the majority of people.In recent years,many researchers began to study traditional Chinese medicine treatment of allergic rhinitis.Mahuang-Fuzi-Xixin Decoction is a classic decoction in Shan Han Lun written by Zhang Zhongjing.It includes three herbs,there are Mahuang,Fuzi and Xixn..Over the years,it is proved clinically,Mahuangfuzixixin Decoction have a significant effect in the treatment of allergic rhinitis.Therefore,the research group of Mahuang-Fuzi-Xixin Decoction studied systematically on the prescription medicinal ingredient active site and the role of anti-allergic rhinitis,found Aconite base and Asarum volatile oil can significantly reduce the histamine content in the blood of the allergic rhinitis model,guinea pig,and also improve local infiltration nasal inflammation;In addition,the active site and the active ingredients of Mahuang-Fuzi-Xixin Decoction herbs can control the inflammation by inhibiting excessive activation of macrophages.Total alkaloids ephedra extract,aconite total alkaloids extract,Asarum decoction,ephedrine,pseudoephedrine,methylephedrine,mesaconitine and Aconitine can simultaneous inhibition the release of IL-4,TNF-?and ?-amino hexosaminidase.But the existing single ingredient or component model cannot effectively expressed pharmacodynamic study medicine multi-component,multi-target,the overall effect of multi-channel,Mahuang-Fuzi-Xixin Decoction herbs studied the material basis pending anti-allergic rhinitis.As a method based on network pharmacology bioinformatics and systems biology,and has been applied to many areas of medical research,such as basic research effective substance,the role of research,traditional Chinese medicine regularity of compatibility mechanism,network pharmacology particularly is suitable to reflect and explain the Chinese multi-component-multi-target effect relationship.In this study,a network pharmacology of Mahuang-Fuzi-Xixin Decoction anti-allergic rhinitis herbs systematic study of the role,forecast Mahuang-Fuzi-Xixin Decoction herbs anti-allergic rhinitis,the main active ingredient groups,mechanism of action and possible indications;then IgE mediated RBL-2H3 cells for the study,the prediction of Mahuang-Fuzi-Xixin Decoction medicinal substances active ingredients preliminary verification;according to preliminary studies,methyleugenol is Asarum the main active ingredient in the volatile oil(content accounted for 22.68%),from the cells and protein levels of methyl eugenol in-depth study of IgE-mediated degranulation of RBL-2H3 cells and its mechanism.From the perspective of the material basis of systematic research on Mahuang-Fuzi-Xixin Decoction herbs anti-allergic rhinitis to explore,clear Mahuang-Fuzi-Xixin Decoction prescription medicine multi-component,multi-target,multi-channel mechanism for the development of the treatment of allergic rhinitis prescriptions provide the basis for new drugs.METHODS1.Network Pharmacology-based study from the herbs of Mahuang-Fuzi-XixinDecoction on treating allergic rhinitis.(1)From Taiwan herbal medicine database(http://tcm.cmu.edu.tw/)and Chinese medicine,pharmacology data platform(http://sm.nwsuaf.edu.cn/lsp/tcmsp.php)and other database,by query and lots the literature review,the chemical composition of the 3 kinds of medical material of Mahuangfuzixixin Decoction was collected.(2)Select the oral bioavailability(oral bioavailability,OB)?30%and drug-like compound(drug-like,DL)?0.1 as potential active ingredients.(3)Use the potential target for drugs forecasting model(DTpre)to predict the target of the active ingredient.(4)Using Cytoscape Software build a "candidate ingredients-candidate target network,""potential component-a potential target for the network," "target-Disease Network",and characteristics of the network(including network degrees,referral number,etc.),predict the material basis of the herbs of Mahuang-Fuzi-Xixin Decoction on treating allergic rhinitis.2.Verify the prediction of ingredients of the herbs of Mahuang-Fuzi-Xixin Decoction.Active ingredients using MTT assay predicted value of RBL-2H3 cells Activity;active ingredient detected by ELISA forecast for IgE-mediated degranulation of RBL-2H3 cells release PGE2,influence LTB4,IL-4,and TNF-?.3.The effect of methyleugenol on IgE-mediated degranulation of RBL-2H3 cells and mechanisms of action.The effect of methyleugenol on RBL-2H3 cells increased the activity MTT assay;detecting methyleugenol for IgE-mediated degranulation of RBL-2H3 cells release of ?-hexosaminidase;and by Western blot detection of methyleugenol for IgE-mediated RBL-2H3 cells arachidonic acid metabolism pathway p-cPLA2/cPLA2,COX-2,p-5-LO/5-LO and IgE/Fc?RI mediated p-Lyn/Lyn,p-Syk/Syk and MAPK family of p-JNK/JNK,p-ERK/ERK,p-P38/P38 protein expression situation.Result1?Network Pharmacology-based study on the herbs from Mahuang-Fuzi-Xixin Decoction on treating Allergic Rhinitis:Through database queries and literature review,367 compounds were collected from herbs of Mahuang-Fuzi-Xixin Decoction,of which 234 from Mahuang,65 from Fuzi,68 from Xixin.The active ingredient screening results show that,in which 54 compounds(14.7%)while meeting the oral bioavailability(OB?30%)and drug-likeness(DL?0.10)conditions,finally 60 compounds from 3 kinds of herbs from Mahuang Fuzi Xixin Decoction were classified as candidate compounds(including ephedrine,pseudoephedrine and other 6 chemical composition have been reported due to possess anti-allergy or regulate immune function,so further targets retain their forecast).The results show that the prediction target,43 candidate compounds with 280 candidate target interact.The compound-the interaction between targets imports Cytoscape construct biological networks through network analysis:a)"candidate ingredients-candidate target" network consists of 322 nodes(43 candidate compound with a target of 280 candidates)and 944 connections.There are 30 compounds and 3 there is a strong interaction between the target protein,which 13 molecules of quercetin,kaempferol,luteolin,methyleugenol,ephedrine,pseudoephedrine and other with six or more target proteins are present action;b)"potential component-a potential target for"network consists of 78 nodes(41 potential compound with 37 potential targets)and 233 connections.And allergic rhinitis-related targets betweenness the top 12 were:cyclooxygenase-2(COX-2,T210),cyclooxygenase-1(COX-1,T209),heat shock protein 90(HSP90,T119),muscarinic acetylcholine receptor M1(CHRM1,T164),?-2 adrenergic receptor(ADRB2,T38),dipeptidyl peptidase 4(DDP4,T84),inducible nitric oxide synthase(NOS3,T182),trypsin 1(PRSS1,T260),tumor necrosis factor(TNF,T262),the transcription factor p65(RELA,T254),5-lipoxygenase(ALOX5,T31)and interleukin 4(IL-4,T146).Network ranking before these targets is 12,just order some adjustments.c)"a potential target-disease" Network Connections 28 targets and 138 kinds of diseases,138 kinds of diseases are divided into tumor diseases accounted for 26.8%,respiratory diseases accounted for 10.1%,cardiovascular disease accounted for 9.42%.2?Prediction of ingredients Mahuang-Fuzi-Xixin Decoction medicinal substances to verify:the model group compared with the normal group,PGE2,LTB4,IL-4 and the release of TNF-? were raised,the difference was statistically significant(P<0.05).Compared with the model group,the ingredients are able to inhibit the release of PGE2,the difference was statistically significant(P<0.05);ephedrine,pseudoephedrine,methylephedrine,methyleugenol,kaempferol,quercetin,luteolin,cotton flavin,apigenin,rutin can inhibit the PGE2,IL-4,the release of TNF-?,the difference was statistically significant(P<0.05);in addition kaempferol,quercetin,luteolin,methyleugenol,cotton flavin,apigenin,rutin can inhibit the release of LTB4,the difference was statistically significant(P<0.05).3?Effect of methyleugenol of IgE-mediated degranulation of RBL-2H3 cells and its mechanism study:Methyleugenol in a concentration range 0-100?M had no effect on the RBL-2H3 cells increase;model group compared with the control group,?-hexosaminidase release were raised,and the difference was statistically significant(P<0.05);25-100?M concentration range of methyleugenol in a concentration-dependent inhibition can IgE-mediated degranulation of RBL-2H3 cells release ?-hexosaminidase,the difference was statistically significant(P<0.05);the model group compared with the normal group,the regulation of inflammation products when 25-1 00?M methyleugenol concentration range of IgE-mediated protein p RBL-2H3 cells-cPLA2,COX-2,p-5-LO,p-Syk,p-JNK,p-ERK,p-P38 display certain regulatory role,and in a dose-dependent manner.Conclusion1?Network pharmacology studies showed the drug,quercetin,kaempferol,luteolin,ephedrine,pseudoephedrine,methyleugenol,cotton Flavin,norepinephrine salsoline and other 13 with six or more of the target protein is present role they are in a very important position,"potential ingredient potential targets," the network may be Mahuang-Fuzi-Xixin Decoction anti-allergic rhinitis medicine Pharmacodynamic material basis.Predicted active molecules mainly adrenergic receptor(adrenergic receptors,ADRs),the biosynthesis of muscarinic receptor(Muscarinic acetycholine receptor)and other neurotransmitters,PGE2,LTB4,IL-4 and other inflammatory mediators key protease(eg,COX-2,5-LOX,etc.)as well as the presence of heat shock protein related targets 90(HSP 90),nitric oxide synthase(NOS),DDPIV,tumor necrosis factor(TNF),trypsin-1,etc.immunomodulatory interaction,thereby explain Mahuang-Fuzi-Xixin Decoction prescription medicines,mainly through anti-inflammatory,immune and other effects to the treatment of the molecular mechanisms of allergic rhinitis;In addition,the network also studied pharmacology Mahuang-Fuzi-Xixin Decoction potential indications and partial results verified by the literature.2?After activation of mast cells can release a variety of inflammatory mediators(such as leukotrienes,tryptase,prostaglandins and interleukins,etc.)to activate the secondary effects downstream target cells-basophils and neutrophils,and ultimately lead to flow tears,nasal itching,sneezing and stuffy nose and other acute allergic reactions.Verification results show that ephedrine,pseudoephedrine,methylephedrine,methyl eugenol,kaempferol,quercetin,luteolin,cotton Flavin,apigenin,rutin for IgE-mediated RBL-2H3 cells PGE2,IL-4,the release of TNF-? is inhibited;additional kaempferol,quercetin,luteolin,methyl eugenol,cotton flavin,apigenin,rutin can inhibit the release of LTB4,indicating that these ingredients may as Mahuang-Fuzi-Xixin Decoction anti-allergic rhinitis medicine pharmacodynamic substances.3?25-100?M concentration range of methyl eugenol may be in a concentration-dependent inhibition of IgE-mediated release of the iconic media ?-hexosaminidase cells after activation RBL-2H3,which exert anti-allergic effect may be related to the regulation of molecular mechanisms arachidonic acid metabolism pathway p-cPLA2/cPLA2,COX-2,p-5-LO/5-LO,IgE/Fc?RI mediated p-Syk/Syk and MAPK family of p-JNK/JNK,p-ERK/ERK,protein expression p-P38/P38 related.