Formononetin protects against ox-LDL-induced endothelial dysfunction by activating PPAR-γ signaling based on network pharmacology and experimental validation

Abstract

Formononetin (FMNT), a flavonoid identified from the Chinese herb Astragalus membranaceus, possesses anti-inflammatory or anti-oxidative properties in different human diseases. This study aims to comprehensively elucidate the function of FMNT in atherosclerosis and its underlying mechanisms. Online public databases were used to identify the drug-disease targets. Proteinprotein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were applied to explore the potential targets and signaling pathways involved in FMNT against atherosclerosis. Human umbilical vein endothelial cells (HUVECs) were exposed to oxidized low-density lipoprotein (ox-LDL) to construct an atherosclerosis cell model in vitro. Endothelial cell function was assessed via examining cell proliferation, inflammatory factors, oxidative markers, reactive oxygen species (ROS), and apoptosis. Western blot was performed to detect the expression of cyclooxygenase-2 (COX-2), endothelial nitric oxide synthase (eNOS), cleaved caspase-3, and peroxisome proliferator-activated receptor-γ (PPAR-γ). A total of 39 overlapping target genes of FMNT and atherosclerosis were identified. Through the PPI network analysis, 14 hub genes were screened and found to be closely relevant to inflammation, oxidative stress, and apoptosis. Results of KEGG pathway assays indicated that lots of targets were enriched in PPAR signaling. Functionally, FMNT could protect against ox-LDL-induced inflammatory reaction, oxidative stress, and apoptosis in HUVECs. Moreover, FMNT attenuated ox-LDL-mediated inactivation of PPAR-γ signaling. GW9662, a PPAR-γ antagonist, reversed the inhibitory effect of FMNT on ox-LDL-induced endothelial injury. In conclusion, FMNT alleviates ox-LDL-induced endothelial injury in HUVECs by stimulating PPAR-γ signaling, providing a theoretical basis for employing FMNT as a potential drug to combat atherosclerosis.

Sitagliptin ameliorates hypoxia-induced damages in endometrial stromal cells: an implication in endometriosis

“…HESCs were seeded on a 96-well microplate and incubated for 24 h. Then 10 μM DCFH-DA dissolved in serum-free medium was added into each well, followed by 30 min incubation. After washing with the serum-free medium, a microplate reader (BMG LABTECH, Offenburg, Germany) was utilized to measure the absorbance at 488/525 nm [ 11 ].…”

Mechanisms of Oxidized LDL-Mediated Endothelial Dysfunction and Its Consequences for the Development of Atherosclerosis

“…There are multiple mechanisms resulting in the endothelial injury triggered by oxLDL, including the excessive ROS production, inflammation, and endoplasmic reticulum (ER) stress. In addition, oxLDL induced cytotoxicity can be reversed by several phytochemicals such as kaempferol, safflower yellow A, oxymatrine, and formononetin ( 16 – 18 ).…”

Pharmacological Mechanism of Ganlu Powder in the Treatment of NASH Based on Network Pharmacology and Molecular Docking

“…Network pharmacology is an emerging discipline proposed by professor Hopkins in 2007 [ 17 ]. This approach breaks the previous concept of a single ingredient/single target/disease and can thus be used as a useful tool in the studies of the “multicomponent, multitarget, and multibiological regulation function synergy” model of Chinese herbs [ 18 , 19 ]. In this study, we studied the potential targets of GLP for NASH using network pharmacology and used the maximum degree value to identify the core targets in the gene network that indicate the role of the nodes in network pharmacology.…”