Rapid Discovery Of Heteropappus Meyendorffii Capitulum Anti-Inflammatory Active Components Based On Spectral-Effect Relationship

Objective:Liquid chromatography-tandem mass spectrometry(LC-MS/MS)was used to determine the constituent absorbed into blood of each polar solvent extract of Heteropappus meyendorffii.The pharmacodynamic study was carried out by establishing a rat model of bacterial pneumonia.Combined with the results of the two methods,the spectrum-efficiency related statistical methods was used to quickly find the "antifebrile" active components in Heteropappus meyendorffii,and the mechanism of action was studied according to the metabolomics technology.Methods:(1)Using the bacterial pneumonia model,96 male rats with SPF-grade SD were divided into 8 groups,including blank group,model group,95% ethanol extract administration group,petroleum ether extract administration group,chloroform extract administration group,ethyl acetate extract administration group,n-butanol extract administration group and roxithromycin positive control group,and the contents of TNF-α,IL-1 and IL-6 were used as the investigation indicators to evaluate the anti-inflammatory activities of different polar solvent extracts of Heteropappus meyendorffii;(2)UHPLC-Q-EXACTIVE-MS/MS technology was used to establish an analysis method for different polar solvent extracts of Heteropappus meyendorffii and drug-containing serum samples after oral administration of Heteropappus meyendorffii in rats.To analyze and compare the chromatograms of control rat serum,contains medicated serum,and different polar solvent extracts of Heteropappus meyendorffii,and according to the retention time,precise molecular weight,secondary fragment ions and other information of each compound,combined with databases and relevant literature,the constituent absorbed into blood in rats after oral administration of different polar solvent extracts of Heteropappus meyendorffii were determined.(3)The Partial least-squares regression analysis method was used to analyze the correlation between the 151 common peak areas of different polar solvent extracts of Heteropappus meyendorffii head tidbits and the three evaluation indexes of anti-inflammatory,and the spectrum-effect relationship of their anti-inflammatory effects was studied.(4)Ultra-performance liquid chromatography tandem quadrupole Orbitrap mass spectrometry was used for analysis,and endogenous differential metabolites with VIP>1 and p<0.05 were screened according to their mass spectra compared with online databases,the change trend of differential metabolites after administration was observed,and finally pathway enrichment analysis was carried out to explain their biological significance.Results:(1)HE results showed that compared with the blank group,the model group could see abnormal lung tissue structure and a large number of inflammatory cell infiltration,and the above symptoms could be alleviated by giving extracts of various polar parts of Heteropappus meyendorffii;The levels of TNF-α,IL-1 and IL-6 in rat serum:Compared with the blank group,the serum concentrations of TNF-α,IL-1 and 1L-6 in the model group were significantly increased(P<0.05),and compared with the model group,the concentrations of TNF-α,IL-1 and IL-6 in the serum of 95% ethanol extract administration group,petroleum ether extract administration group,chloroform extract administration group,ethyl acetate extract administration group,n-butanol extract administration group and positive control group decreased significantly(P<0.05);(2)UHPLC-Q-Exactive-MS/MS technology was used to identify 626 constituents absorbed into blood,of which 151 components had a degree of fit greater than 80% and a deviation value less than 5.Terpenoids accounted for(16.4%),followed by alkaloids(12.5%),phenylpropanoids(12.5%),flavonoids(6.6%),phenols(4.6%)and quinones(2.6%);(3)Based on the spectrum-effect relationship to study the content of various chemical components and their anti-inflammatory effects,pharmacodynamic studies have shown that the main anti-inflammatory components in Heteropappus meyendorffii are indole-3-carbaldehyde,guanine,benzoic acid,succinic acid,ethyl caffeate,isozineepidine,adipic acid,galactobionate,resveratrol,ceteroline,cetanedioic acid,p-cresol,heptanedioic acid,piperidicalic acid,etc;(4)Metabolomics results showed that after administration of extracts from polar parts of Heteropappus meyendorffii in pneumonia model rats,the differential metabolites in the body showed different degrees of callback,which were related to valine,leucine and isoleucine biosynthesis,glycine,serine and threonine metabolism,and the n-butanol extract group had the most differential metabolites that could be moderated.Conclusions : From the pharmacodynamic results,the extracts of various polar parts of Heteropappus meyendorffii had therapeutic effects on rats with bacterial pneumonia model.Serum Pharmacochemistry were used to study the medicinal substances of Heteropappus meyendorffii,and the research ideas of application spectrum-effect relationship were used to elaborate the basis of anti-inflammatory active substances of Heteropappus meyendorffii.On this basis,metabolomics was used to explore the molecular mechanism of anti-inflammatory effect of Heteropappus meyendorffii,and it was found that Mongolian medicine Heteropappus meyendorffii regulated amino acid metabolism disorders to exert anti-inflammatory effect.