1.Analysis of Pharmacodynamic Material Basis and Action Mechanism of Anemarrhenae Rhizoma-Phellodendri Chinensis Herb Pair in the Treatment of Type 2 Diabetes before and after Salt Processing Based on Serum Pharmacochemistry and Net-work Pharmacology
Xiangqi MENG ; Chunling ZHANG ; Yaoxia JIA ; Jinjie LEI ; Yangmiao XIA ; Rui TAN ; Lingying YU
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(5):479-493
OBJECTIVE To analyze the serum chemical composition of rats after intragastric administration of water extract of crude Anemarrhenae Rhizoma-crude Phellodendri Chinensis and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis based on liquid chromatography-mass spectrometry(LC-MS)technology,predict the effect of salt processing on the treatment of type 2 dia-betes in Anemarrhenae Rhizoma-Pheellodendri Chinensis combined with network pharmacology,and preliminarily verify it through in vitro experiments.METHODS Rats were continuously intragastrically administered with crude Anemarrhenae Rhizoma-crude Phello-dendri Chinensis drug pair and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis drug pair water extract twice,with an in-terval of 1 h.After 60 min of the last administration,the blood was taken from the abdominal aorta,and the protein was precipitated by methanol.After dissolution,the chromatographic column was Shim-pack GIST C18(4.6 mm×150 mm,5 μm);the mobile phase A was 0.1%formic acid water,and the mobile phase B was 0.1%formic acid-acetonitrile;gradient elution,positive and negative ion full scan mode,mass scan range 100-1 500 m/z.Combined with the secondary spectrum of the database and literature,the blood compo-nents of crude Anemarrhenae Rhizoma-crude Phellodendri Chinensis drug pair and salted Anemarrhenae Rhizoma-salted Phellodendri Chinensis drug pair were analyzed and identified.The disease targets of type 2 diabetes were retrieved,and the protein interaction net-work analysis,GO and KEGG pathway enrichment analysis were performed on the intersection targets of blood components and diseases.The"blood components-targets"network diagram was constructed,and the selected core components and core targets were verified by molecular docking using AutoDock software.In the verification experiment,HepG2 cells were used as the experimental ob-ject,and the insulin resistance model was induced by high insulin and high glucose.CCK8 method was used to test the effect of Rhizo-ma Anemarrhenae-Phellodendri Chinensis on cell proliferation before and after salt processing.Western blot was used to detect the ex-pression of PI3K-AKT signaling pathway-related proteins.RESULTS 15 prototype components and 1 mangiferin metabolic compo-nent were identified in the serum of rats.17 prototype components and 1 mangiferin metabolite were identified in the rat serum of the water extract of Anemarrhenae Rhizoma-Phellodendri Chinensis after salt processing.The contents of mangiferin,berberine and 3-isobutylglutaric acid in the blood components after salt-processing were higher than those in the raw products.According to the results of KEGG and GO,the treatment of type 2 diabetes may be related to the transcriptional regulation of RNA polymerase,inflammatory re-sponse,AGE-RAGE,PI3K-AKT pathway and insulin resistance.Cell experiments showed that the ratio of p-PI3K/PI3K,p-AKT/AKT and GLUT4 protein expression could be up-regulated before and after salt processing,and the effect of salt processing group was better than that of the crude group.CONCLUSION This experiment preliminarily revealed the components of Anemarrhenae Rhizo-ma-Phellodendri Chinensis drug pair entering the blood before and after salt exposure,and suggested that ferulic acid,berberine,ber-berrubine,mangiferin,mTOR,SIRT1,EGFR and PPARA may be the main components and targets of Rhizoma Anemarrhenae-Phel-lodendri Chinensis after salt processing to enhance the therapeutic effect of type 2 diabetes.The mechanism may be to enhance the role of PI3K-AKT and other related signaling pathways,providing an important reference for the pharmacodynamic material basis and clini-cal application of Anemarrhenae Rhizoma-Phellodendri Chinensis before and after salt processing.
2.GC-MS Analysis of Effect of Different Processing Conditions on Irritation of Magnoliae Officinalis Cortex
Pingjun CAI ; Mengqin QIANG ; Yangmiao XIA ; Yaoxia JIA ; Xiangqi MENG ; Jinjie LEI ; Fang LIU ; Lingying YU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(16):197-206
ObjectiveTo investigate the key compounds affecting the irritation and to clarify the effect of heating and the addition of ginger juice as the auxiliary material during the processing on the irritation of Magnoliae Officinalis Cortex(MOC) by comparing the irritation and composition of volatile oil in MOC and its different processed products. MethodVolatile oil in raw products, water-processed products, ginger-dried products, ginger-fried products(the amounts of ginger were 10%, 50%, respectively) of MOC were extracted by steam distillation and subjected to rabbit eye irritation experiment, and the volatile components of each sample were detected by gas chromatography-mass spectrometry(GC-MS). Principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to analyze the data of each sample by SIMCA 14.1. The relative contents of different processed products were compared two by two with those of and raw products or ginger-fried products, and the markers that might be related to the irritation were sorted out according to the principles of variable importance in the projection(VIP) value >1 and P<0.05, and the factors influencing the differences in irritation were analyzed. ResultCompared with the blank group, the administration groups all had irritation to the eyes of rabbits, and the degree of irritation was in the order of raw products>water-processed products>ginger-dried products>ginger-fried products(10%)>ginger-fried products(50%). The results of PCA and OPLS-DA showed that there were differences in the volatile oil from raw products and different processed products. According to VIP value>1 and P<0.05, and combined with the results of eye irritation experiment, ten volatile compounds related to irritation changes were screened out. Among them, cis-cinnamaldehyde was only detected in raw products, the relative contents of β-caryophyllene, (+)-delta-cadinene, α-humulene, γ-muurolene, (-)-isoledene and citral all increased to different degrees, the contents of p-cymene, 1(10)-4-cadinadien-15-ol and β-eudesmol all decreased to different degrees. ConclusionThe irritation of MOC is reduced after heating and processing with ginger juice, and the synergistic effect of both is more effective for reducing irritation. Among the differential markers associated with changes in irritation, the increase in the relative content of citral is closely related to the addition of ginger juice, while the decrease in the relative contents of cis-cinnamaldehyde, p-cymene, 1(10)-4-cadinadien-15-ol is related to heating, and the changes of other components may be related to the synergistic effect of heating and ginger juice.