Quality Markers Between Crude And Wine-processed Scutellaria Baicalensis Based On "Components-targets-metabolism"

Scutellariae radix(SR)is one of the most popular traditional Chinese medicines with the effects of heat-clearing and detoxification,wine processing increased the inclination and direction of SR’s actions,thereby strengthening its efficacy in clearing the upper-energizer lung damp heat.In this study,to clarify the difference of its material basis,the chemical components were studied between crude Scutellariae Radix(CSR)and wine-processed Scutellariae Radix(WSR).Then,to explore the processing mechanism of SR from an overall perspective,the metabolomics was adopted to investigate the effects of CSR and WSR on pneumonia to clarify the levels of potential biomarkers and the differences in the mechanism of them.Finally,combined with network pharmacology,a strategy of "component-target-metabolism" was proposed to predict potential quality markers before and after Scutellariae Radix based on the correlation analysis.Details are shown as follows:1.The chemical components of CSR and WSR were identified by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS).According to the information of retention time,accurate molecular weight and secondary fragment ion,a total of 53 chemical compounds were detected.Principal component analysis(PCA)and partial least-squares discriminant analysis(PLS-DA)were performed to discriminate the chemical profiles of CSR and WSR.Then,taking the content of the chemical markers as an importance index,a contribution function was introduced to screen chemical markers.The results show that 17 compounds were identified as discriminatory chemicals between CSR andWSR,and 10 components,namely,baicalein,skullcapflavone II,wogonoside,baicalin,scutevulin-7-O-glucuronide,hispidulin-7-O-glucuronide,oroxylin A-7-O-glucuronide,hispidulin,chrysin-7-O-glucuronide,and skullcapflavone I,were identified as chemical markers with an accumulative content contribution of 88.75%.In addition,this work revealed the best wine-processed time was 18 min by investigating the changes of chemical markers in SR during processing.2.Pneumonia model was established by oral-pharyngeal inhalation of lipopolysaccharide(LPS).To explore the effects of CSR and WSR,the contents of interleukin-6(IL-6),interleukin-8(IL-8),tumor necrosis factor alpha(TNF-α),and the contents of nitric oxide(NO)were detected.The results show that WSR exhibited a more remarkable effect on improving pneumonia than CSR by reducing the levels of inflammatory factors,including NO,TNF-α,IL-6,and IL-8.Next,On the basis of UPLC-QTOF/MS technology,an unequal curative effect was revealed by nontargeting metabolomics.Sixteen biomarkers were discovered in the plasma of LPS-induced rats.After WSR treatment,the disturbed metabolic profiling,tryptophan,alpha-linolenic acid,pregnenolone,9(S)-HPODE,and retinoate were restored to control-like levels.The levels of phytosphingosine,sphinganine,SM(d18:1/22:1(13Z)),and palmitoleic acid in the CSR group were closer to those in the normal control group.Pathway analysis indicated that CSR acted on pneumonia by regulating the abnormal sphingolipid metabolism pathways.However,an WSR-mediated cure of pneumonia was linked primarily to reversing the abnormality of retinol metabolism pathways and tryptophan metabolism pathways.3.The "Component-target-pneumonia" network was constructed,and 127 related protein targets were obtained.The relevant targets were analyzed by Go enrichment.It was found that the biological process of Go enrichment also included the three inflammatory signaling pathways involved in the metabolomics study.Among which the targets of tryptophan metabolism pathway were CYP1B1,CYP1A1,CYP1A2,MAOA and MAOB.The related targets of retinol metabolism pathway were CYP1A1,CYP1A2,UGT1A1,CYP2C9 and CYP3A4.The related targets of sphingolipid metabolism pathways were NOS3,ADORA3,TNF,MAPK14,ABCC1,PIK3 CA,TP53,MAPK1,NFKB1 and MAPK3.Combining the results of network pharmacology,metabolomics and correlation analysis,oroxylin A-7-O-glucuronide baicalin were regarded as the Q-markers of CSR;baicalein and wogonin were considered as the Q-markers of WSR.In summary,this work investigate the changes of material basis and therapeutic effect of CSR and WSR with modern technology.Elucidated the processing mechanism of Scutellariae Radix.Predicted potential quality markers before and after Scutellariae Radix based on the correlation analysis.From the perspective of holistic of traditional Chinese medicine,taking the excavation of quality markers of different processed products as the ultimate objective,to establish a quality control and evaluation system and provides ideas for the quality control of decoction pieces of Chinese medicine.