Active Components of Pueraria lobata through the MAPK/ERK Signaling Pathway Alleviate Iron Overload in Alcoholic Liver Disease.

OBJECTIVE: To delve into the primary active ingredients and mechanism of Pueraria lobata for alleviating iron overload in alcoholic liver disease. METHODS: Pueraria lobata's potential targets and signaling pathways in treating alcohol-induced iron overloads were predicted using network pharmacology analysis. Then, animal experiments were used to validate the predictions of network pharmacology. The impact of puerarin or genistein on alcohol-induced iron accumulation, liver injury, oxidative stress, and apoptosis was assessed using morphological examination, biochemical index test, and immunofluorescence. Key proteins implicated in linked pathways were identified using RT-qPCR, western blot analysis, and immunohistochemistry. RESULTS: Network pharmacological predictions combined with animal experiments suggest that the model group compared to the control group, exhibited activation of the MAPK/ERK signaling pathway, suppression of hepcidin expression, and aggravated iron overload, liver damage, oxidative stress, and hepatocyte death. Puerarin and genistein, the active compounds in Pueraria lobata, effectively mitigated the aforementioned alcohol-induced effects. No statistically significant disparities were seen in the effects above between the two groups receiving drug therapy. CONCLUSION: This study preliminarily demonstrated that puerarin and genistein in Pueraria lobata may increase hepcidin production to alleviate alcohol-induced iron overload by inhibiting the MAPK/ERK signaling pathway.

Integrating network pharmacology and experimental verification to explore the mechanism of puerarin against oliguria in acute alcoholism.

Purpose: This study was designed to evaluate the pharmacological mechanisms of puerarin against oliguria in acute alcoholism via network pharmacology analysis combined with experimental verification. Methods: First, this study established an acute alcoholism rat model, compared the changes in urine volume in each group, and observed the therapeutic effect of puerarin by H&E staining, biochemical, RT-qPCR, and immunohistochemical analyses. Second, puerarin-related targets were searched in TCMS, PubChem, CNKI, Wanfang, PubMed, and GeenMedical Academic databases. Also, potential disease targets were obtained from the GeneCards, MalaCards, and NCBI-gene databases and genes with puerarin target gene intersections were screened out. The interaction network for co-predicted targets was obtained using the STRING database, and the core targets were imported into Cytoscape for visualization using DAVID Bioinformatics Resources 6.8. The essential genes were subjected to the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathway enrichment analyses to predict related biological processes and significant signaling pathways. Finally, molecular docking was used to examine the interaction of puerarin with key targets, and the core targets were validated further by RT-qPCR and Western blotting. Results: Compared to the model group, the urine volume of the rats was significantly increased after puerarin treatment, and the levels of anti-diuretic hormone (ADH) and aquaporin 2 (AQP2) expression were decreased. Searching the intersection of puerarin and acute alcoholism targets yielded 214 potential targets, 837 biological processes, and 185 signaling pathways involved. The molecular docking results indicated a good affinity between puerarin and key targets (cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), cAMP-response element-binding protein (CREB), and c-Fos). RT-qPCR and Western blotting further verified that puerarin could down-regulate the expression of cAMP/PKA/CREB/c-Fos. Conclusion: This study identified the potential targets of puerarin against oliguria in rats with acute alcoholism using network pharmacology and animal experiments. The mechanism may be closely related to the cAMP signaling pathway.

Detection of Gegen Adulteration Using Multiple Fingerprints Coupled With Chemometric Strategy.

BACKGROUND: The root of Pueraria montana var. lobata (gegen) is a well-known traditional Chinese medical herb, which is prone to be accidentally contaminated with Pueraria montana var. thomsonii, Pueraria wallichii, and Pueraria peduncularis due to the morphological character similarity. These adulterations might cause quality confusion and safety issues. OBJECTIVE: In this study, the screening technique to detect adulteration in gegen was developed using multiple fingerprints and chemometrics. METHOD: A range of gegen samples and possible known adulterants including Pueraria montana var. thomsonii, Pueraria wallichii, and Pueraria peduncularis were collected. FT-IR and HPLC coupled with different chemometric techniques, including similarity analysis (SA), hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least-squares discriminant analysis (OPLS-DA), provide the qualitative chemometric models for gegen adulteration detection. RESULTS: FT-IR and HPLC combined with OPLS-DA successfully differentiated authentic gegen from adulterants. Both FT-IR and HPLC units can be used as alternative methods to traditional methods. The HPLC showed better performance in identifying samples than FT-IR. CONCLUSIONS: The use of FT-IR and HPLC coupled with chemometrics could potentially be the proper selection method for the early quality evaluation of gegen. This method can be used to combat fraud in the herbal industry in the future. HIGHLIGHTS: FT-IR and HPLC combined with chemometrics analysis were developed to discriminate between Pueraria lobata (gegen) and adulterants. The multiple fingerprints combined with multivariate methods were successfully applied to the study of the gegen and its adulterants. The chemometrics analysis using SA and OPLS-DA indicate significant differentiation in the chemical composition of these species. This research provides important chemotaxonomic references in species identification.