1.Analysis of Chemical Composition in Puerariae Thomsonii Stem Base and Investigation of Its Effect on Liver Metabolites in Mice Based on UPLC-Q-TOF-MS/MS
Lingbang MENG ; Huan YU ; Xiaoying WU ; Yijing GUO ; Fei QU ; Qianfeng GONG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):174-182
ObjectiveTo systematically compare the chemical compositional differences between Puerariae Thomsonii stem base(PTSB) and Puerariae Thomsonii Radix(PTR), and to explore the potential hepatoprotective effects of PTSB by liver metabolomics. MethodUltra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to analyze the chemical compositions of PTSB and PTR. Twenty Kunming mice aged 6-8 weeks, half male and half female, were randomly divided into the blank group(sterile water) and PTSB group(1.95 g·kg-1), with 10 mice in each group, and the drug was administered by gavage for 14 d, and the body mass was weighed once a day. After the last administration, mice were anesthetized, organs such as heart, liver, spleen, lungs and kidneys were collected, and the organ index was calculated. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), total cholesterol(TC) and triglyceride(TG) in the serum of mice from each group, the morphological changes of heart, liver, spleen, lung and kidney tissues were observed by hematoxylin-eosin(HE) staining, and the regulation of PTSB for the hepatic metabolic profiles of mice was analyzed by UPLC-Q-TOF-MS/MS, then the differential metabolites between the blank group and PTSB group were designated, and the metabolic pathways was enriched by Kyoto Encyclopedia of Genes and Genomes(KEGG). ResultA total of 19 common chemical constituents were identified from PTSB and PTR, all of which were the main pharmacodynamic substances of PTR. The pharmacodynamic results showed that PTSB could control the growth of body mass of mice and reduce the contents of TC, TG, ALT and AST in serum of mice. HE staining observations and organ indexes showed that there was no significant effect of PTSB on all major organs at the highest clinically equivalent dose. A total of 38 differential metabolites were identified by metabolomics, of which 35 were up-regulated and 3 were down-regulated. These differential metabolites were mainly compounds such as amino acids, fatty acids, vitamins, steroids, nucleosides, pyrimidines and alkaloids. Three key metabolic pathways, including tyrosine metabolism, vitamin B6 metabolism and tryptophan metabolism, were screened by metabolic pathway analysis. ConclusionPTSB has a similar chemical composition to that of PTR, and it may regulate the metabolism of amino acids and vitamins through the flavonoids and isoflavonoids, thus exerting a potential hepatoprotective effect. This study provides an experimental reference for the clinical application and product development of PTSB.
2.Dynamic Changes of Volatile Components During Processing Process of Jianchangbang Characteristic Ginger Processed Xingpo Pieces Based on HS-GC-MS
Yuxun CHEN ; Wanai XU ; Jingyu WU ; Meiqi LU ; Lingbang MENG ; Cheng XU ; Hailian ZHENG ; Xian ZHANG ; Na ZHANG ; Qianfeng GONG ; Huan YU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):127-134
ObjectiveTo reveal the influence of Jianchangbang characteristic processing method on the change process of volatile components and the processing mechanism of reducing toxicity and increasing efficiency of Magnoliae Officinalis Cortex(MOC) by studying the changes in the composition and content of volatile components during the processing of ginger processed Xingpo pieces. MethodSamples of raw products, ginger juice moisturized products and stir-fried and heap moisturized products of MOC were taken according to the set time points, and headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to determine the contents of volatile components in the samples, and the relative content of each component was obtained by peak area normalization. Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were performed on the sample data using SIMCA 14.1 software, and the differential components during the processing were screened with variable importance in the projection(VIP) value>1 as the indicator. ResultA total of 68 volatile components were identified in the samples, among which some of the chemical components with similar structures showed similar trends of changes, and there was also the phenomenon of interconversion between compounds. Compared with the raw products, the contents of 42 components in ginger juice moisturized products increased, while the contents of 25 components decreased, 19 components were unique, and 4 components were unique to the raw products. Compared with ginger juice moisturized products, MOC in the early stage of piling had three unique components, and the contents of 11 components such as cyclosativene and (+)-α-pinene increased, and the contents of 5 components such as tricyclic terpene and α-curcumene decreased, and ginger juice moisturized products had four unique components. Compared with the early stage of piling, in the later stage, the contents of 8 components such as (+)-α-pinene and camphene significantly increased, while the contents of 6 components such as linalool and α-selinene significantly decreased. During the processing of MOC, there were significant changes in the chemical composition of the samples before and after 20 days. The differences between ginger juice moistening and the early stage of piling, the early stage and the later stage of piling could be clearly distinguished. ConclusionDuring the preparation process of ginger processed Xingpo pieces, the addition of ginger juice can reduce the contents of stimulating components, and the contents of active components continue to increase in several stages, such as the addition of ginger juice, frying and heap moisturizing, the quality of the decoction pieces may change significantly at about 20 d of processing. This study can provide a research basis for exploring the processing mechanism of ginger processed Xingpo pieces.