Background: Ginsenoside Re (Re) has been shown to activate small-conductance calcium-activated potassium (SKCa) current in human coronary artery endothelial cells (HCAECs). We aimed to investigate whether Re increased SKCa current via glucocorticoid receptor (GR), its non-genomic pathway phosphoinositide 3-kinase-protein kinase B (PI3K-Akt/PKB), and endothelial nitric oxide synthase (eNOS), and whether SKCa mediated Re-induced increase in nitric oxide (NO), prostacyclin (PGI2), epoxyeicosatrienoic acid (EET), and/or hydrogen peroxide (H2O2).
Methods: Whole-cell patch clamp technique was employed to study Re-activated HCAEC currents, using specific inhibitors of the proposed mediating pathway. NO and H2O2 were assayed with colorimetric methods; PGI2 and EET were investigated using ELISA. eNOS phosphorylation was assessed using Western blot analysis.
Results: Re (1 μM) significantly increased HCAEC whole-cell current at +80 mV to 173.73 ± 43.90 % (mean ± SD). Apamin (SKCa blocker) could virtually eliminate Re-induced current and apamin-insensitive current could not be increased by Re, while blockers of other endothelial potassium channels did not produce the same effects. Moreover, antagonists of GR, PI3K, Akt/PKB, and eNOS effectively prevented Re's action. Re-induced eNOS phosphorylation and NO production could be prevented by blockers of SKCa, GR, or Akt/PKB, but Re-induced PGI2 production could not be prevented by apamin, while EET and H2O2 were not increased by Re.
Conclusion: Re enhances SKCa current and NO production via GR-PI3K-Akt/PKB and eNOS activation; in turn, SKCa current is essential for Re-increased NO. However, Re-induced PGI2 release is independent of SKCa current. These findings could facilitate further research about ginseng effects on coronary artery and possible use in cardiovascular diseases.
Background: Hepatic fibrosis (HF) continues to be a significant global health concern, substantially contributing to morbidity and mortality due to the absence of effective therapeutic options. This study examines the pharmacological effectiveness and underlying mechanisms of Notoginsenoside R2 (R2) in mitigating HF, aiming to find a new multifunctional candidate for therapeutic application.
Methods: An integrative methodology utilizing network pharmacology, molecular docking, and experimental validation was implemented. In vitro models (HSC-T6), in vivo systems (zebrafish), and microinjection of morpholinos were employed to corroborate the antifibrotic effects of R2 and transcription 3 (STAT3)-dependent processes.
Results: Network pharmacology identified 32 common targets between R2 and HF, with a particular emphasis on pathways critical for the activation of HSCs. Molecular docking confirmed strong interactions between R2 and signal transducer and activator of STAT3. In vitro, R2 inhibited HSCs proliferation and decreased the expression of α-SMA, COL-I, Desimin and TIMP1. In vivo, R2 mitigated thioacetamide-induced fibrosis in zebrafish, leading to decreased collagen deposition and suppression of pro-inflammatory cytokines. Mechanistically, R2 induced senescence in HSCs via the STAT3 pathway, characterized by increased expression of cyclin-dependent kinase inhibitor 2A (CDKN2A/p16) and cyclin-dependent kinase inhibitor 1A (CDKN1A/p21), as well as components of the senescence-associated secretory phenotypes (SASPs).
Conclusion: This study identified R2 as a regulator of STAT3 with dual antifibrotic effects: reduction of the inflammatory microenvironment and induction of senescence. These findings position R2 as a viable treatment candidate for HF, necessitating additional clinical investigation.
Background: Colorectal cancer is the third most common cancer worldwide and the fourth leading cause of cancer death. Protopanaxatriol (PPT), one of the main active metabolic ginsenosides of ginseng, has been found to have neuroprotective and anti-inflammatory effects, but its role in regulating colon cancer development remained unclear.
Purpose: We sought to confirm the inhibitory effect of PPT on colon cancer cells and elucidate its target and mechanism.
Methods: MTT assay, colony formation, invasion, migration assays, cell apoptosis, and cell-cycle analysis were performed. Quantitative real-time PCR, Western blotting, bioinformatic analysis using the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology, and cellular thermal shift assay experiments were also employed.
Results: PPT can inhibit the cloning, migration, invasion, and proliferation of colon cancer cells. PPT can increase the number of apoptotic bodies, promote the expression of the apoptotic proteins caspase-9 and caspase-3, induce apoptosis, and inhibit cell proliferation. In addition, by regulating cyclin, PPT can increase the expression of p21 and p27 proteins and inhibit the expression of the cyclin D1 protein, thereby inhibiting the G1/S transformation in the cell cycle. We further demonstrated that PPT can target AKT, reduce its protein expression, and reduce tumor progression and the expression of inflammatory factors caused by AKT high expression (TNF-α, IL-1β, and IL-6), thereby playing a role in inhibiting colon cancer progression.
Conclusion: We are the first to demonstrate that the ginsenoside PPT can inhibit the activity of colon cancer cells by directly binding AKT.
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