Regularity Of Compound Herbal Formula For Pneumonia Treated By Professor Wang Shou-chuan And Anti-RSV Effect Of Jinxin Oral Liquid And Baicalein Based On Metabonomics

BackgroundTraditional Chinese medicine has great advantage of treating pediatric pneumonia.Professor Wang treats pediatric pneumonia well,with high safety and low side effects.Thus,medication rules in treating pneumonia should be analyzed.ObjectiveTo study Professor Wang Shou-chuan’s medication rules in treating pneumonia through data mining method.Methods249 cases of prescription information from 119 patients treated by Professor Wang Shou-chuan were collected for cluster and association analysis with SPSS 19.0 and Clementine 12.0 software with the purpose of mining Professor Wang’s academic experiences in treating pneumonia.Results(1)Cluster analysis is divided into four categories,Cl:Ephedra,Bitter almonds,Radix Scutellariae,Polygonum cuspidatum,Semen lepidii,Perilla Fruit and Radix peucedani;C2:Mulberry leaves,Morus alba,Bile Arisaema,loquat leaves,Flos Magnoliae and Rhizoma paridis;C3:Radix Platycodonis,Citron fruit,Fritillaria thunbergii,Earthworm,Periostracum cicada,Fructus Trichosanthis,Radix Asteris,Radix stemonae,Gypsum and salvia miltiorrhizae;C4:Astragalus membranaceus,atractylodis macrocephalae,Saposhnikovia divaricata,Os Draconis Preparata,Concha Ostrear Preparata.(2)Association rules analysis:from the relationship between the two drugs,the Radix peucedani and Ephedra,Ephedra and Bitter almonds,Mulberry leaves and Morus alba,Radix peucedani and Bitter almonds,Perilla Fruit and Semen lepidii,Perilla Fruit and Radix peucedani,Radix peucedani and Radix Scutellariae,Polygonum cuspidatum and Radix Scutellariae,Bile Arisaema and Radix Scutellariae,Bile Arisaema and Radix peucedani,Morus alba and Radix Scutellariae,and other commonly used drugs are combined.From the three drugs-related point of view,the basic embodiment of the treatment of pneumonia are releasing and suppressing lung qi,clearing heat and resolving phlegm.Drugs that can release lung qi include Ephedra and Mulberry leaves,and that can suppress lung qi include Bitter almonds,Morus alba,Folium eriobotryae,Radix peucedani,Semen lepidii and Perilla Fruit,and that clearing heat drugs include Radix Scutellariae and Polygonum cuspidatum,and drugs resolving phlegm include Bile Arisaema and Bitter almonds.ConclusionMedication rules in treating pneumonia of professor Wang can be revealed preliminarily using the cluster analysis and association rules analysis.The core drugs contain Ephedra,Bitter almonds,Mulberry leaves,Morus alba,Radix Scutellariae,Polygonum cuspidatum,Semen lepidii,Perilla Fruit,Bile Arisaema,Radix peucedani,with compatible drugs Radix Platycodonis,Citron fiuit,Fritillaria thunbergii,Fructus Trichosanthis,Radix Asteris,Radix stemonae and Gypsum to clear heat and resolve phelgm to supress qi of lung and relieve cough。BackgroundPneumonia is a common respiratory disease in pediatrics,and can cause serious damage to children’s health,which is listed as one of three important pediatric diseases around the world by the world health organization.It is estimated that viral infection accounted for about 1/3 in children with acute lower respiratory infection,and is on the rise.As one of the most important pathogens,respiratory syncytial virus can cause infant capillary bronchitis,bronchial pneumonia,and interstitial pneumonia,etc.At present,the pathogenesis of RSV infection still remains not fully clear,and safe and effective drugs or vaccines still lack.With the characteristic of its treatment based on syndrome differentiation,soft effect and less side effects,traditional Chinese medicine has certain advantages in the treatment of children’s respiratory disease.Professor Wang has lead our team engaged in traditional Chinese medicine treatment of pneumonia for almost twenty years,and developed the Jin xin oral liquid which is safe and effective and can open the inhibited lung-energy,clear heat-evil,dissipate phlegm and relieve cough and asthma.Extensive clinical and experimental studies have shown that it can cure the RSV infection and regulate immune function.As an important effective component of Jinxin oral liquid,baicalein has shown a broad spectrum antiviral activity.However,the most experimental studies may have many defects,such as single,independent,asynchronous indicators.Therefore,systematic and holistic research technical approach should be introduced for comprehensive analysis.Metabonomics is the important component of systems biology.When buoyed by a variety of external factors,organisms represent response and regulation which can be detected instanly,sensitively and really using metabonomics.As a kind of system research method,the changes of endogenous metabolites after stimulation or disturbance can be detected to explore overall the mechanism of action of drug.Thus,Metabonomics is adopted to explain the mechanism of the antiviral effect of Jinxin oral liquid and baicalein.PurposeTo detect the metabolic characteristic and the changes of endogenous metabolites in plasma,urine,bronchoalveolar lavage fluid,spleen tissue of BALB/c mice and cell with the GC-MS and UPLC/MS technology,to explore the machanism of the antiviral effect of Jinxin oral liquid and baicalein.MethodBALB/c mice were challenged intranasally to establish the RSV pneumonia models and treated by Jin xin oral liquid and baicalein respectively.Plasma,urine,bronchoalveolar lavage fluid,spleen tissue were collected.lung tissue pathological changes were observed.Cell were cultured and infected by RSV for 1.5 hour,then treated by baicalein for 24 hour and quenched with liquid nitrogen.water-methanol-chloroform mixture to extract intracelluar metabolites.After preprocessed,plasma,urine,spleen tissue and cell samples were detected with GC-MS technology,and bronchoalveolar lavage fluid were detected with UPLC/MS.PCA and OPLS-DA analysis were conducted with SIMCA-P.Potential biomarkers were selected and related metabolic pathways were analyzed to explain the mechanism of antiviral effect of Jinxin oral liquid and baicalein.Result1.Pathological changes in lung tissues of mice infected with RSV were mainly as interstitial pneumonia,multifocal inflammatory cell infiltrated,which maily were mononuclear cells and lymphocytes,along with mild hyperemia(+)and thickening of alveolar walls.After administration of Jinxin oral liquid and baicalein,the congestion,edema and inflammatory infiltration in lung tissue alleviated to some extent.2.Metabonomics of Jinxin Oral Liquid(1)Plasma:OPLS-DA showed that the plasma of three groups were separated completely without overlap,and the the parameters R2Y=0.934,Q2=0.889.Six potential biomarkers were identified and selected,among which were lactic acid,urea,glutamine,1,5-andro-sorbitol,glucose,arachidonic acid.(2)OPLS-DA showed that the plasma of three groups were separated completely without overlap,and the the parameters R2Y=0.85,Q2=0.746.Four potential biomarkers were identified and selected,among which were lactic acid,glycine,Stearic acid and palmitic acid.(3)OPLS-DA showed that the plasma of three groups were separated completely without overlap,and the the parameters R2Y=0.959,Q2=0.895.Six potential biomarkers were identified and selected,among which were L-proline,L-glutamic acid,valine,urea,hypoxanthine,glucose.(4)OPLS-DA showed that the plasma of three groups were separated completely without overlap,and the the parameters R2Y=0.972,Q2=0.924.Six potential biomarkers were identified and selected,among which were L-carnitine,propionyl carnitine,adenosine,dihydrogen nerve sheath ammonia alcohol,leukotriene D5 and glucuronic acid estrone.3.Metabolomics of baicalein(1)OPLS-DA showed that the cell of three groups were separated completely without overlap,and the the parameters R2Y=0.974,Q2=0.917.Five potential biomarkers were identified and selected,among which were serine,L-aspartic acid,L-glutamic acid,citric acid,and glycine.(2)OPLS-DA showed that the plasma of three groups were separated completely without overlap,and the the parameters R2Y=0.912,Q2=0.84.Six potential biomarkers were identified and selected,among which were lactic acid,urea,glycine,glutamine,1,5-andro-sorbitol and glucose.(3)OPLS-DA showed that the urine of three groups were separated completely without overlap,and the the parameters R2Y=0.917,Q2=0.767.Four potential biomarkers were identified and selected,among which were lactic acid,malic acid,palmitic acid.and stearic acid.(4)OPLS-DA showed that the BALF of three groups were separated completely without overlap,and the the parameters R2Y=0.954,Q2=0.828.Five potential biomarkers were identified and selected,among which were glutamate,carbonyl carnitine,adenosine,glycosidase acyl sheath ammonia alcohol,leukotriene D5.Conclusion1.The mechanism of antiviral effect of Jinxin oral liquid and baicalein could be explored with UPLC/MS and GC-MS technology.2.RSV infection could cause metabolic disorder in the plasma,urine,BALF and spleen tissues of mice,mainly including amino acids,organic acids,and fatty acids.3.RSV infection could be treated by Jinxin oral liquid and baicalein.4.Jin xin oral liquid and baicalein could regulate the metabolic disorder caused by RSV infection.The antiviral effect of Jinxin oral liquid is mainly concerned with glutamate metabolism,alanine,aspartic acid,and glutamic acid metabolism,valine,leucine,and isoleucine,and Arachidonic acid metabolism,glycine,serine and threonine metabolism,arginine and proline metabolism,and pyruvate metabolism.Baicalein is mainly related with glutamine metabolism,alanine,glutamic acid and aspartic acid metabolism,glycine,serine and threonine metabolism.