Liu, S.-P. | bioRxiv Science

bioRxiv2024

Explore the mechanism of Zigui Yichong Formula in reducing the apoptosis of ovarian granulosa cells in premature ovarian insufficiency based on network pharmacology, molecular docking and cell experiments

ObjectiveThis research is conducted with the objective of exploring the underlying mechanism by which the Zigui Yichong Formula (ZGYCF) diminishes granulosa cell apoptosis in the context of premature ovarian insufficiency (POI), utilizing network pharmacology, molecular docking, and cellular experimentation approaches. MethodsThe active constituents and potential therapeutic targets of the 12 medicinal herbs in ZGYCF, which include Rehmannia glutinosa, Cervus nippon, Cornus officinalis, Ligustrum lucidum, Lycium barbarum, Paeonia lactiflora, Astragalus membranaceus, Codonopsis pilosula, Atractylodes macrocephala, Angelica sinensis, Cyperus rotundus, and Glycyrrhiza uralensis, were identified through searches in the TCMSP, BATMAN, HERB, and ETCM databases. Targets associated with the POI condition were gathered from the OpenTargets, DrugBank, and GeneCards databases. Subsequently, a Venn diagram illustrating the compound-target-disease interaction was generated to derive a set of common targets that bridge the gap between pharmacological and pathological targets. A drug-component-target-disease network diagram was created using Cytoscape 3.9.1. Additionally, protein-protein interaction (PPI) networks were built utilizing the STRING database and visualized with Cytoscape to pinpoint key targets within the overlapping target set. Functional annotation and pathway enrichment analyses, including GO and KEGG pathway analyses, were performed using the clusterProfiler package in R 4.2.1 to investigate the underlying mechanisms by which the drug may influence the disease state. The molecular docking of pivotal active constituents with central targets was carried out using AutoDock Tools. Following this, in vitro studies were executed to corroborate the anticipated mechanisms of action of ZGYCF on POI that were inferred from the network pharmacology analysis. ResultsThe selected active components include quercetin, kaempferol, and {beta}-sitosterol. The core targets identified are Tp53, Bcl-2, and Caspase-3. GO functional and KEGG enrichment analyses indicate that these core targets are primarily enriched in the p53 signaling pathway. Molecular docking results show that quercetin, kaempferol, and {beta}-sitosterol have good binding affinity with TP53, Bcl-2, and Caspase-3. Additionally, in vitro experiments demonstrate that ZGYCF medicated serum can reduce ACR-induced apoptosis in KGN cells, increase Bcl-2 expression, and decrease the expression of p53, Bax, Caspase-3, and the Bcl-2/Bax ratio. ConclusionZGYCF exerts therapeutic effects on POI through multiple targets and pathways, and it may reduce ACR-induced apoptosis in KGN cells by modulating the p53 signaling pathway. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=130 HEIGHT=200 SRC="FIGDIR/small/614279v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@1e55e2forg.highwire.dtl.DTLVardef@18a67d0org.highwire.dtl.DTLVardef@1a596d0org.highwire.dtl.DTLVardef@503fde_HPS_FORMAT_FIGEXP M_FIG C_FIG

pharmacology and toxicology↗

bioRxiv2022

Whether the ADP-ribosyltransferase activity of Ta-sro1, a noncanonical PARP protein, contributes to its function in salinity-stress tolerance?

ADP-ribosylation mediated by ADP-ribosyltransferases (ARTs) is an intricate modification that regulates diverse cellular processes including DNA repair, chromatin remodeling and gene transcription responding to stresses. In addition to the canonical poly(ADP-ribose) polymerases (PARPs), plant specific SRO (Similar to RCD One) family also contain the catalytic core of the PARP domain. However, whether the PARP domains in SROs execute the ART function is still under debate. In 2014, we reported a wheat SRO, Ta-sro1, had the ADP-ribosyltransferase activity and enhanced wheat seedling growth and abiotic stress resistance, however, a recent work by Vogt et al. showed that Ta-sro1 without ADP-ribosyltransferase activity. Based on the recent progress on PARPs and SROs in relation to ADP-ribosyltransferase activity, along with our former and recent evolving results, we argued that Ta-sro1 is a non-canonical ADP-ribosyltransferase with the enzymatic activity. Although we have revealed the novel mechanism of Ta-sro1 regulate redox homeostasis and enhance salinity stress tolerance through interacting with TaSIP1, it is of interest to further clarify whether and how the enzymatic activity of Ta-sro1 responsible for the salinity tolerance of wheat. Our study raises some interesting points and caveats that helpful for understanding the research progresses and debates about the enzymatic activity of SROs.

molecular biology↗