Akt x BAX x Bcl-2 x PI3Ks

Senegenin is a major bioactive aglycone derived from Polygala tenuifolia Willd. (Polygalaceae), a traditional Chinese medicinal herb ("Yuan Zhi") widely used for the treatment of cognitive impairment and central nervous system disorders. Although senegenin has been reported to exert neuroactive and anti-inflammatory effects, the intracellular mechanisms underlying its regulation of microglia-driven neuroinflammation remain incompletely understood.

This Study aimed to characterize the effects of senegenin on inflammatory responses in lipopolysaccharide (LPS)-stimulated microglia and to elucidate the molecular mechanisms underlying its anti-inflammatory activity, with particular emphasis on Akt and the cGAS-STING pathway.

An integrative network pharmacology strategy was employed to predict senegenin-associated molecular targets and signaling pathways, followed by mechanistic validation in LPS-stimulated N9 microglial cells. Oxidative stress, mitochondrial function, DNA damage, inflammasome activation, and inflammatory signaling were assessed using flow cytometry, immunofluorescence, immunoblotting, and biochemical assay.

Network pharmacology analysis identified 110 overlapping targets, including CASP3, PIK3CA, and HIF1A, with significant enrichment in the PI3K-Akt and HIF-1 signaling pathways. Experimental validation demonstrated that senegenin markedly attenuated LPS-induced reactive oxygen species production, preserved mitochondrial membrane potential (Δψm), and reduced mitochondrial and nuclear DNA damage. These effects were associated with restoration of Akt phosphorylation and coordinated suppression of the cGAS-STING-TBK1-IRF3 signaling axis, as evidenced by reduced phosphorylation of STING, TBK1, and IRF3, along with diminished IFN-β expression. Senegenin further inhibited NF-κB/NLRP3 inflammasome priming and activation, shifted apoptotic signaling toward cell survival (decreased Bax and increased Bcl-2), and reduced the expression of pro-inflammatory mediators, including TNF-α, IL-6, IL-1β, COX-2, and iNOS.

This Study demonstrates that senegenin attenuates LPS-induced microglial inflammatory activation by limiting oxidative stress and DNA damage, thereby suppressing cGAS-STING-dependent inflammatory signaling and associated cell death pathways, potentially through Akt phosphorylation. Although confined to a cellular model, these findings provide mechanistic insight into the ethnopharmacological basis of senegenin and establish a foundation for future in vivo investigations into its neuroprotective relevance in neurological disorders.

To explore the inhibitory effects of mulberry leaf flavonoid extract (MFE) on high glucose-induced diabetic encephalopathy (DE) in a cell model, and to investigate the underlying mechanisms combined with network pharmacology prediction and experimental validation.

A DE cell model was established by chronic high glucose stimulation of HT22 mouse hippocampal neuronal cells. Following intervention with MFE, cell viability, advanced glycation end products (AGEs) content, and ferroptosis-related indices were detected. Network pharmacology was used to screen potential signaling pathways and key targets of MFE, while Western blotting validated the expression of proteins related to the screened pathways and targets.

Network pharmacology screening identified Akt1, ALB, TNF, IL-1B, and IL-6 as the top-ranked targets. Molecular docking confirmed stable binding between major MFE components and these potential targets. MFE treatment markedly upregulated ferritin heavy chain (FTH) and ferroportin (FPN) levels while decreasing transferrin receptor (TFRC) levels, attenuating iron overload in DE cells. It also suppressed AGEs accumulation, which activated the downstream PI3K/Akt/Nrf2/SLC7A11/GPX4 ferroptosis regulatory pathway to mitigate oxidative stress and inhibit ferroptosis. Additionally, MFE activated Akt, leading to p38 MAPK deactivation, reduced Bax/Bcl-2 ratio, and decreased Caspase-3 levels, blocking the apoptosis pathway. It alleviated inflammatory injury by suppressing the p38 MAPK/NF-κB pathway, thereby reducing TNF-α, IL-1β, and IL-6 protein levels.

MFE inhibits ferroptosis by upregulating the PI3K/Akt/Nrf2/GPX4 signaling pathway. This study provides valuable insights into the therapeutic application of MFE for DE.

Premenopausal women diagnosed with cancer may suffer from ovarian toxicity, which affects their quality of life. Cisplatin (CIS) is an effective chemotherapeutic drug used for different types of tumors. However, CIS has been reported to cause ovarian toxicity. Aberration of the PI3K/AKT signaling has been implicated in CIS-induced ovarian toxic effects. Piceatannol (PIC) is a naturally occurring polyphenolic stilbene that has garnered significant research interest due to its known antioxidant, anti-inflammatory, and anti-apoptotic activities. The aim of this study was to investigate the potential protective role of PIC against CIS-induced ovarian toxicity in female rats. Thirty female rats were divided randomly into five groups, and treated for 17 days. (1) control, (2): PIC (10 mg/kg), (3): CIS (6 mg/kg), (4): low dose PIC (5 mg/kg) + CIS (6 mg/kg), and (5): high dose PIC (10 mg/kg) + CIS (6 mg/kg). Pretreatment with PIC significantly prevented the CIS-induced histopathological alterations, inhibited follicles loss, and inhibited the decrease in serum AMH. PIC exhibited antioxidant activity by significantly preventing MDA production and the depletion of GSH and antioxidant enzymes in ovarian tissues in a dose-dependent manner. PIC markedly decreased the immunostaining expression of iNOS, TNF-α, and NF-κB in ovarian tissues. Furthermore, PIC significantly attenuated immune-expression of PTEN and enhanced those of PI3K and phosphor-AKT in ovarian tissues of CIS-treated rats. Finally, PIC modulated mRNA expression of Bcl-2, BAX and CASP3 in favor of anti-apoptosis. PIC protects against CIS-induced ovarian toxicity by exerting antioxidant, anti-inflammatory, and anti-apoptotic effects, primarily through modulation of the PTEN/PI3K/p-AKT axis.