The Multi-targets Networks Construction And Effect On Severe Asthma Induced By PM2.5 Of San-ao Decoction And Its Added Formulae

Objective:This study is to comb the understanding of asthma in Chinese medicine,the development and research status of San-ao Decoction.This study is to establish the multi-targets networks of the active components of San-ao Decoction and its added formulae,and compared to the network of asthma to reveal the potential targets of San-ao Decoction and its added formulae.Then PM2.5 is used as the stimulating factor in OVA-sensitized and challenged mice to exacerbate asthmatic effects.On the basis of this model,this paper will investigate the effects and mechanisms of San-ao Decoction and its added formulae for treating asthma.Method:1.The origin of disease,pathogenesis,syndrome differentiation and treatment in Chinese medicine literature is explored.The high-frequency drugs in modern asthma formulae is calculated.Development process of San-ao Decoction is reviewed.Then the clinical curative effects,active components and effect mechanisms of San-ao Decoction and its added formulae are combed in modern literature.2.The multi-targets networks of San-ao Decoction and its added formulae for treating asthma are construct by network pharmacology.Then the potential targets of San-ao Decoction and its added formulae are explored.3.The air pollutant fine particulate matter(PM2.5)is introduced into OVA-sensitized and challenged mice.PM2.5 is administrated at two doses,1.6mg·kg-1 or 8.0mg·kg-1 in OVA-challenged stage to establish 28 and 42 day asthma models.Then the changes in airway resistance(RI)and lung compliance(Cdyn)in response to acetylcholine chloride(ACH)are evaluated,and blood,bronchoalveolar lavage fluid(BALF)and lung tissue samples are taken.Number of eosinophils in blood and various leukocytes in BALF are determined.Lung protein is extracted and probed by immunoblot for TRPA1,TRPV1 expression.Levels of IL-13,SP,PGD2 and NGF in BALF are measured by ELISA.The ideal model are used to explore the effects and mechanisms of San-ao Decoction and its added formulae for treating asthma.4.On the basis of the construct of multi-targets networks and the exacerbating asthma model induced by OVA plus PM2.5,the effects of San-ao Decoction and its added formulae on PM2.5-induced asthma mice are investigated in improving airway inflammation,airway hyperresponsiveness and lung histopathology.Levels of IL-13,SP,PGD2 and NGF in BALF are measured by ELISA.The expressions of TRPA1,TRPV1,TRPV4 and TRPM8 mRNA in lung tissue are detected by q-PCR.Relevant TRPs protein expressions are detected by Western blot.Result:1.In modern cold-asthma and hot-asthma formulae,the higher frequency drugs were ephedra sinica,semen armeniacae amarum,glycyrrhiza uralensis and so on.According to the records by ancient doctors of different dynasties,San-ao Decoction and its added formulae were applied with cough and asthma.The main compatibility for San-ao Decoction are adding herbs with the function of stimulating sweat secretion,reducing phlegm,relieving cough and asthma,clearing heat,interior-warming,astringent and qi regulating.San-ao Decoction and its added formulae mainly intervened in asthma from the aspects of inhibition of inflammation,regulation of immunity,regulation of nerve-receptor and airway responsiveness in the modern researches.2.Network pharmacology study showed that 11 active components in San-ao Decoction involved 74 targets and 47 pathways,16 active components in Wu-ao Decoction involved 98 targets and 71 pathways,asthma involved 82 targets and 52 pathways.MAPK(Mitogen-activated protein kinase)was the core target for San-ao Decoction.MAPK and GTPase were the core targets groups for Wu-ao Decoction.And-TNF(Tumor necrosis factor)was the core target for asthma.Targets groups of neuronal ligand-receptor interaction pathway were the potential targets for asthma.San-ao Decoction,Wu-ao Decoction and asthma showed the characteristics of multi-targets,multi-channels and integrated regulation.More enzymes proteins and more widely targets and channels were found in San-ao Decoction and Wu-ao Decoction comprared to those in asthma.3.PM2.5 exacerbated the changes in inflammatory cells,especially EOS in blood and BALF,the degree of change in RI and Cdyn in response to ACH,and lung histopathology,as indicated by inflammatory cell infiltration,thickened bronchial smooth muscles and bronchial mucosa damage.In addition,higher expression of TRPA1,TRPV1 protein,IL-13,SP,PDG2 and NGF were observed in mice exposed to PM2.5 along with OVA.All effects were most pronounced in PM2.5 mice in the 42 day asthma model.4.San-ao Decoction and Wu-ao Decoction could improve the symptoms in asthma mice induced by OVA plus PM2.5,especially in the high dose groups of the two formulae,such as reducing the number and ratio of inflammatory cells(especially EOS)in blood and BALF,reducing lung airway hyperresponsiveness,improving lung injury.In addition,the high expression of TRPA1,TRPV1 mRNA and protein,IL-13,SP,PDG2 and NGF were decreased in San-ao Decoction and Wu-ao Decoction.Compared to OVA plus PM2.5 group,the high dose of San-ao Decoction significantly reduced the expression of TRPA1 protein,IL,SP,PDG2 and NGF(P<0.01).The expression of TRPV1 protein,IL-13,SP,NGF(P<0.01)and TRPA1 protein and PDG2(P<0.05)were significantly decreased in the high dose group of Wu-aoDecoction.Conclusion:1.This paper explored the origin of disease,pathogenesis,syndrome differentiation and treatment in Chinese medicine literature.In modern cold-asthma and hot-asthma formulae,the higher frequency drugs are ephedra sinica,semen armeniacae amarum,glycyrrhiza uralensis and so on.According to the records by ancient doctors of different dynasties,San-ao Decoction and its added formulae were applied with cough and asthma.The main compatibility for San-ao Decoction are adding herbs with the function of stimulating sweat secretion,reducing phlegm,relieving cough and asthma,clearing heat,interior-warming,astringent and qi regulating.The modern literature showed that San-ao Decoction and its added formulae mainly intervened in asthma from the aspects of inhibition of inflammation,regulation of immunity,regulation of nerve-receptor and airway responsiveness.2.Network pharmacology study showed that San-ao Decoction,Wu-ao Decoction and asthma had the characteristics of multi-targets,multi-channels and integrated regulation.More enzymes proteins and more widely targets and channels were found in San-ao Decoction,Wu-ao Decoction compared to those in asthma.This paper showed that 11 active components in San-ao Decoction involved 74 targets and 47 pathways,16 active components in Wu-ao Decoction involved 98 targets and 71 pathways,and asthma involved 82 targets and 52 pathways.MAPK was the core target for San-ao Decoction.MAPK and GTPase were the core targets groups for Wu-ao Decoction.And TNF was the core target of asthma.Targets groups of neuronal ligand-receptor interaction pathway were the potential targets for asthma.It showed the relationship of commonness and individuality between the basic and added formulae.The potential targets of nerve-receptor pathway in asthma would become the new research direction of San-ao Decoction and Wu-ao Decoction.3.On successful establishment of asthma model induced by OVA plus PM2.5 and the network screening results of the two formulae,this study investigated the effects and the potential nerve-immunological mechanisms of San-ao Decoction and Wu-ao Decoction.They could improve the symptoms of asthma mice,especially in the high dose groups of the two formulae,such as reducing the number and ratio of inflammatory cells(especially EOS)in blood and BALF,reducing lung airway hyperresponsiveness,improving lung injury.San-ao Decoction and Wu-ao Decoction could regulate TRP channels and the release of neurotransmitters.San-ao Decoction was focused on inhibition of TRPA1 channel,while Wu-ao Decoction had the effect on inhibition of TRP A1 and TRPV1 channels.