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Background. Xiezhuo Huayu Yiqi Tongluo Formula (XHYTF) consists of 14 Chinese herbal medicines. In this study, we investigated the potential mechanism of XHYTF in the treatment of uric acid nephropathy (UAN) through network pharmacology, molecular docking, and in vivo methods. Methods. Using various pharmacological databases and analysis platforms, information on the active ingredients and targets of Chinese herbal medicine was collected, and UAN disease targets were retrieved using OMIM, Gene Cards, and NCBI. Then common target proteins were integrated. A Drug-Component-Target (D-C-T) map was constructed to screen core compounds and build a protein-protein interaction (PPI) network. Further, Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for common targets, and a Drug-Component-Target-Pathway (D-C-T-P) network diagram was constructed. The molecular docking simulation was performed to verify the binding affinity between core components and hub targets. Subsequently, the UAN rat model was established, followed by the collection of serum and renal tissues. The expression levels of indicators in the serum were determined using an enzyme-linked immunosorbent assay. The pathological changes of renal tissues were detected using H & E staining and Masson staining. The expression of related proteins in renal tissue was detected by western blot. Results. In the study, 216 active ingredients and 439 targets in XHYTF were screened, and 868 targets were identified as being related to UAN. Among them, 115 were common targets. Based on the D-C-T network, quercetin, luteolin, β-sitosterol, and stigmasterol were observed to be the key active ingredients of XHYTF that were effective against UAN. The analysis of the PPI network revealed TNF, IL6, AKT1, PPARG, and IL1β as the 5 key targets. GO enrichment analysis revealed that the pathways were mainly concentrated in cell killing, regulation of signaling receptor activity, and other activities. Subsequently, KEGG pathway analysis revealed that multiple signaling pathways, including the HIF-1, PI3K-Akt, IL-17, and other signaling pathways, were closely related to the action of XHYTF. All 5 key targets were confirmed to interact with all core active ingredients. In vivo experiments indicated that XHYTF significantly reduced blood uric acid and creatinine levels, alleviated inflammatory cell infiltration in kidney tissues, reduced the levels of serum inflammatory factors such as TNF-α and IL1β, and ameliorated renal fibrosis in rats with UAN. Finally, western blot revealed decreased levels of PI3K and AKT1 proteins in the kidney, which confirmed the hypothesis. Conclusion. Collectively, our observations demonstrated that XHYTF significantly protects kidney function, including alleviation of inflammation and renal fibrosis via multiple pathways. This study provided novel insights into the treatment of UAN using traditional Chinese medicines. [ABSTRACT FROM AUTHOR]

Zanthoxylum armatum DC. (Z. armatum) is an herbal medicine with various active ingredients and pharmacological effects. However, modern studies found that Z. armatum is hepatotoxic. The liver is the target organ for toxic effects and an important site for lipid metabolism. The effects of Z. armatum on lipid level and metabolism in the liver are still unclear. This study aimed to analyze hepatic lipid levels, lipid metabolites and metabolic pathways of action of Z. armatum based on lipidomics, to investigate the potential hepatotoxic mechanism of Z. armatum. Different doses (62, 96, and 150 mg/kg) of the methanolic extract of Z. armatum (MZADC) were administered to ICR mice by gavage. The hepatotoxicity of MZADC was assessed by the liver index, serum biochemical measurements, and histopathological examination. Lipid levels measured by the serum lipid index were evaluated in the mice. Lipidomics was used to screen for differential lipid metabolism markers and lipid metabolism pathways in the liver. Western blot analysis was performed to investigate the effects of MZADC on the liver. Liver index values and serum alanine transaminase and aspartate transaminase levels were increased in the MZADC group. Histopathology examination revealed hepatocyte necrosis, watery degeneration of the hepatocytes, and hepatic cord rupture in the livers of mice. Serum levels of low-density lipoprotein cholesterol, cholesterol, and triglycerides were elevated, and high-density lipoprotein cholesterol levels were decreased. Lipidomics screening for markers of differential lipid metabolism in the liver, and altered profiles of differential metabolites indicated that glycerophospholipid metabolism, linoleic acid metabolism, alpha-linolenic acid metabolism, glycosylphosphatidylinositol-anchored biosynthesis, sphingolipid metabolism and arachidonic acid metabolic pathways were significantly associated with MZADC-induced liver injury. Western blots confirmed that the protein expression of LC3, Beclin-1, ATG5, ATG12 and ATG16L1 was decreased, and p62 was increased in the MZADC group. The proportion of p-PI3K/PI3K and p-AKT/AKT was increased. The liver injury induced by MZADC involved many different lipid metabolites and lipid metabolic pathways, which may be related to autophagy. This study provides a new perspective on the hepatotoxicity study of Z. armatum and provides a reference for the safe application of Z. armatum in the medicine and food fields. [Display omitted] • Based on lipidomics, this study revealed hepatotoxicity induced by Zanthoxylum armatum DC. • The metabolites of liver lipid difference induced by Zanthoxylum armatum DC. may serve as potential biomarkers. • Glycerophospholipid metabolism pathways were significantly associated with Zanthoxylum armatum DC. induced liver injury. [ABSTRACT FROM AUTHOR]

Aconitum species, with a medicinal history of 2000 years, was traditionally used in the treatment of rheumatism, arthritis, bruises, and pains. However, many studies have reported that Aconitum species can cause arrhythmia in experimental animals, resulting in myocardial fibrosis and cardiomyocyte damage. Cardiotoxicity is the main toxic effect of aconitine, but the detailed mechanism remains unclear. This study aimed to explore the effects and underlying mechanism of autophagy in H9c2 cardiomyocytes induced by aconitine. H9c2 cells were incubated with different concentrations of aconitine for 24 h, and the intervention sections were pretreated with various inhibitors for 1 h. The effects of aconitine on the oxidative DNA damage, autophagy and viability of H9c2 cells were evaluated by flow cytometry, confocal microscopy, enzyme-linked immunosorbent assay and Western blot. In H9c2 cells, the cell viability declined, LDH release rate, the number of autophagosomes, protein expression levels of LC3 and Beclin-1 increased significantly after 24 h of aconitine incubation. The pretreatment of autophagy inhibitor 3-MA decreased markedly autophagosomes and protein expression levels of LC3 and Beclin-1, which suggested that aconitine could induce cell autophagy. The significant increase of ROS and 8-OHdG showed that aconitine could cause oxidative DNA damage through ROS accumulation. Meanwhile, treatment of aconitine dramatically increased AMPKThr172 and ULK1Ser317 phosphorylation, and Compound C inhibited AMPKThr172 and ULK1Ser317 phosphorylation, which proved that aconitine induced autophagy via AMPK activation mediated ULK1 phosphorylation. Antioxidant NAC significantly reduced LDH, ROS and 8-OHdG, inhibited the phosphorylation of AMPKThr172 and ULK1Ser317, and down-regulated autophagosomes and proteins expression levels of LC3 and Beclin-1. Consequently, the inhibition of oxidative DNA damage and AMPK/ULK1 signaling pathway alleviated the aconitine-induced autophagic death of H9c2 cells. These results showed that aconitine induces autophagy of H9c2 cardiomyocytes by activating AMPK/ULK1 signaling pathway mediated by oxidative DNA damage. The autophagy induced by aconitine in cardiomyocytes is dependent on the activation of the AMPK pathway, which may provide novel insights into the prevention of aconitine-related toxicity. [Display omitted] [ABSTRACT FROM AUTHOR]