Tandem mass tag-based proteomics analysis reveals the effects of Guri Gumu-13 pill on drug-induced liver injury.

The incidence of drug-induced liver injury (DILI) is second only to viral hepatitis and steatohepatitis in China, and DILI has become a serious public health problem that cannot be ignored. Guri Gumu-13 pill (GRGM) is a traditional Chinese medicine (TCM), which has a protective effect on liver diseases. However, the underlying therapeutic mechanisms of GRGM for DILI are still vague. In this study, the protective effect of GRGM on acetaminophen (APAP)-induced DILI was investigated based on the proteomics clues. The effects of GRGM on APAP-induced DILI in rats were studied using tandem mass tag (TMT)-based quantitative proteomics technology. Besides, western blotting was exerted to verify related proteins. Using the TMT-based quantitative proteomics approach, 237 proteins were identified as regulated in APAP-induced DILI and 58 proteins were regulated by GRGM. The 17 overlapping differentially expressed proteins (DEPs) were identified, and 7 proteins were inversely regulated. Enrichment analysis of KEGG indicated that metabolic pathways, linoleic acid metabolism, and retinol metabolism might be affected in DILI. Next, Cyp2c11, Aldh1a1, and Fads2 were verified with molecular biotechnology. GRGM exerts therapeutic effects through multi-pathways regulation in the treatment of DILI. This work may provide proteomics clues for the continuation of research on DILI treatment with GRGM.

Magnetic solid-phase extraction based on magnetic amino modified multiwalled carbon nanotubes for the fast determination of seven pesticide residues in water samples.

A rapid and simple analytical method based on magnetic solid-phase extraction with magnetic amino modified multiwalled carbon nanotubes with ultra-high performance liquid chromatography-tandem mass spectrometry is reported for the determination of seven pesticides (futriafol, metalaxyl, myclobutanil, napropamide, epoxiconazole, fipronil and diniconazole) in water samples. In this study, magnetic amino modified multi-walled carbon nanotubes were synthesized and selected as a new kind of material to adsorb pesticides in the water samples. Various magnetic solid-phase extraction parameters, such as the amount and type of adsorbent, extraction methods, extraction time, the type and volume of desorption solvent, desorption time and solution ionic strength, were systematically optimized. Under optimum conditions, the method validation results showed good linearity and recoveries. The calibration curves were in the range of 1.0-100 ng mL-1 for napropamide, epoxiconazole, metalaxyl, and fipronil, while they were 5.0-500 ng mL-1 for futriafol, myclobutanil, and diniconazole, with determination coefficients (R2) higher than 0.9909. The limits of quantification were 1.0-5.0 ng mL-1 and the limits of detection were 0.3-1.5 ng mL-1. The recoveries of the seven pesticides ranged from 80.4% to 103.2%. This developed method, which is more convenient and effective in comparison with traditional methods, has been successfully applied for the analysis of pesticides in water samples qualitatively and quantitatively.

Effects of mongolian medicine Terminalia chebula Retz. on 6 CYP450 enzymes in rats.

Terminalia chebula Retz. (TCR) is a medicinal material commonly used in Mongolian medicine. After consulting the literature at home and abroad, current research on TCR focuses on chemical composition, pharmacodynamics, and fingerprints. The pharmacokinetics of TCR has not been reported. Cytochrome P450 (CYP450) is the main drug-metabolizing enzyme, and its activity may be induced or inhibited by certain drugs, resulting in drug interactions in clinical applications. The objective of this study was to establish a high performance liquid chromatography (HPLC) method that can simultaneously detect multiple probe drugs to determine the effect of TCR on the activities of CYP450 enzymes CYP2C19, CYP2E1, CYP2D6, CYP2C9, CYP3A4, and CYP1A2. Wistar rats (male) were divided into 5 groups according to the randomization principle, namely the control group, the positive group, and the high, medium and low dose group. After 15 days of continuous administration, the mixed probe drug was injected into the vein, and then a small amount of blood was collected from the orbital vein at different time points. After the samples were processed, the blood concentration of each probe drug was measured by the established HPLC method. The pharmacokinetic parameters of each probe drug were calculated using DAS software. Compared with the control group, the plasma clearance (CL) of chlorzoxazone and omeprazole decreased, and the maximum plasma concentration (Cmax) and area under the curve (AUC) increased in the TCR group. The pharmacokinetic parameters of theophylline, midazolam, metoprolol, and tolbutamide did not differ significantly. The results indicated that TCR mainly inhibited the activities of CYP2E1 and CYP2C19, but had no effect on the activities of CYP1A2, CYP2C9, CYP3A4 and CYP2D6. Extra care should be taken when drugs metabolized by CYP2C19 and CYP2E1 enzymes are used in combination with TCR, as drug-herb interactions may occur. These results can guide the clinical application of related drugs and provide valuable information for drug interactions. The main component that affects enzyme activity may be tannins in the water extract.