Metabonomics Research On Volatile Oils Of Raw Angelica And Its Different Processed Products Intervenein LPS Inflammation Rats Based On LC-Q/TOF-MS

Rats were intraperitoneally injected with lipopolysaccharide(LPS) to establish systemic inflammation model. The anti-inflammatory function of volatile oils of Angelica(VOAS) and its different processed products were evaluated by detecting the liver and lung histopathologic slide and the change of related cytokines and inflammatory medium of rats in control group(Control), inflammation model group(Model), the volatile oils of Angelica(S-VOAS), parching Angelica with wine(J-VOAS), charred Angelica(C-VOAS), parching Angelica with soil(T-VOAS), parching Angelica with oil(O-VOAS) and dexamethasone positive control group(Dex). On this basis, LC-Q/TOF-MS was applied to detect the metabolites in plasma, liver, lung and urine of rats in different group. The method of partial least-squares discriminant analysis(PLS-DA) was used as multivariate analysis to filter different metabolites. Combined the high resolution MS spectrometry in Mass Hunter software with the metabolite database searches: HMDB(http://www.hmdb.ca), Mass Bank(http://www.Massbank.jp), Metlin(http://metlin.Scripps.edu) and KEGG(http://www.genome.jp/k:egg) to initially identify the potential biomarkers. Then, the identified biomarkers were put into Met PA(http://metpa.metabolomics.ca) to build metabolic pathways. By analyzing the relationship among anti-inflammation related metabolites, metabolic pathways and cytokines, we studied the anti-inflammatory mechanism and difference of Angelica and its different processed products on LPS –induced inflammation rats from the overall aspects. The main results were as follows:1. Volatile oils of Angelica and its different processed products and dexamethasone could down-regulate white blood cells(WBC) and neutral grain cell percentage value(NE%) in blood of LPS-induced inflammation rats, hepatocyte fatty degeneration and lung inflammatory cell infiltration, which indicated that volatile oils of Angelica and its different processed products could effectively inhibit inflammation symptoms and protect the liver and lung injury of rats. Compared with control group, the content of TNF-α, IL-6, IL-lβ, IL-2, IL-10 and NO increased significantly(P<0.05) in inflammation model group. Compared with inflammation model group, the content of TNF-α, IL-6, IL-lβ, IL-2, IL-10 and NO decreased significantly(P<0.05) in J-VOAS, C-VOAS, T-VOAS, O-VOAS and S-VOAS. Therefore, the anti-inflammatory mechanism of Angelica and its different processed products was speculated to directly or indirectly inhibit LPS target cells secreting pro-inflammatory cytokines and inflammation medium, promote anti-inflammatory cytokine release.2.The plasma, liver, lung and urine samples were detected by metabolomics technology based on LC-Q/TOF-MS. Combined with multivariate analysis PLS-DA, the samples in control group and inflammation model group completely separated, and J-VOAS, C-VOAS, T-VOAS, O-VOAS and the S-VOAS were close to control group and positive control group, in which J-VOAS and C-VOAS the most nearest. This indicated that inflammation model was replicated successfully and violate oils of Angelica and its different processed products were able to intervene the inflammatory response of rats. In addition, 21, 16, 25 and 13 biomarkers related to inflammation were screened in plasma, liver, lung and urine. These biomarkers were mainly involved in glutamate and glutamic acid ester metabolism, glutathione metabolism, taurine and hypotaurine metabolism, arachidonic metabolism, galactose metabolism, linoleic acid metabolism, glycine, serine and threonine metabolism and biotin metabolism. With the intervention of violate oils of Angelica and its different processed products, the content of these biomarkers returned to normal levels and the disturbed metabolic pathways were improved.3. In the PLS-DA matrix graph and histogram of biomarkers of plasma, liver, lung and urine of rats, the J-VOAS and C-VOAS showed the better anti-inflammatory effect, while there was some difference in J-VOAS and C-VOAS on regulating the abnormal metabolism. C-VOAS showed better regulating effect in glutamate and glutamic acid ester metabolism, glutathione metabolism, taurine and hypotaurine metabolism, valine, leucine and isoleucine metabolism and TCA cycle, which include the malic acid, linoleic acid, glutathione, glutamate, taurine, citric acid, valine, galactose, alanine and biotin. However, J-VOAS showed better regulating effect in phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism, glycine, serine and threonine metabolism, pentose and glucuronate tautomerism, which included tryptophan, pyruvate, glycine, threonine, phenylalanine and tyrosine.To sum up, violate oils of Angelica and its different processed products exerted anti-inflammatory activity by regulating inflammation-related biomarkers, the metabolic pathways and the dynamic balance among cytokines. The anti-inflammatory effect of J-VOAS and C-VOAS was better.