BAX x Bcl-2 x COX-2

Hepatocellular carcinoma (HCC) is a major public health problem, with a poor prognosis in patients with advanced disease. We aimed to investigate the cytotoxic activity of coumestrol against human hepatocellular carcinoma HepG2 cells. Crystal violet (CV) assay was performed for cell cytotoxicity assessment on Vero cells (normal Kidney) and HepG2 cells. Apoptosis was analyzed by Annexin V/FITC staining. The relative expression of BAX, bcl-2, NF-Kβ, PCNA, MMP2, Caspase-3, Caspase-9, COX2, and MMP9 was detected using quantitative real-time PCR (qPCR). In addition, immunohistochemical analysis (IHC) of Bax, and PCNA were established. Results indicated that coumestrol induced significant toxicity in HepG2 cells. Annexin V-FITC staining assays revealed that coumestrol-induced cytotoxicity in HepG2 cells was mediated through apoptosis stimulation. The apoptosis in HepG2 cells was mediated through caspase-activation. Cell invasion was inhibited by coumestrol in HepG2 cells via inhibition of MMP-2 and MMP-9 expressions. IHC confirmed the strong expression of Bax and nuclear expression of PCNA in treated cells. To the best of our knowledge, limited studies have investigated the impact of coumestrol. The study showed that coumestrol exhibited cytotoxic and apoptotic effects against HepG2 cells, accompanied by inhibition of invasion-related gene expression.

Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality, largely due to multidrug resistance (MDR). Pioglitazone (PIO) and simvastatin (STAT), originally used for metabolic disorders, also modulate oncogenic pathways. This study evaluated a nanotechnology-based co-delivery approach to improve their therapeutic efficacy against chemoresistant HCC.

Lipid nanoparticles (LNPs) encapsulating PIO and STAT (PSLNPs) were fabricated and functionalized with chitosan (CPSLNPs) or folate-conjugated chitosan (FCPSLNPs). Physicochemical properties (size, zeta potential, polydispersity) were characterized. Hemolysis and MTT assays assessed safety and cytotoxicity in HepG2 cells. qPCR quantified IL-1β, IL-6, BCL2, and BAX expression. Molecular docking examined drug interactions with key proteins involved in proliferation, ferroptosis, inflammation, and MDR. All formulations exhibited nanoscale size and uniform dispersion. Chitosan modification increased zeta potential, and stability studies confirmed consistent physicochemical properties. Hemolysis assays demonstrated biocompatibility. Compared with free drugs, PSLNPs and their functionalized forms significantly enhanced cytotoxicity in HepG2 cells. Gene expression analysis showed downregulation of IL-1β, IL-6, and BCL2, with upregulation of BAX, indicating enhanced apoptosis. Docking studies revealed STAT strongly interacted with SDH, GST, HDAC1, and JNK3, while PIO showed higher affinity for COX-2, GPX4, HO-1, and MMP-9, supporting complementary multi-target effects.

Functionalized PSLNPs improved the delivery and efficacy of PIO and STAT, overcoming MDR by modulating multiple oncogenic pathways. These results highlight their promise as a multi-targeted nanotherapeutic strategy for HCC treatment.

Sodium arsenite (SA) is a potent carcinogenic compound which is evident to prompt multiple organs damage including the heart. Azaleatin (AZA) is a novel therapeutic agent with excellent biological properties.

The current investigation was executed to evaluate the mitigative efficacy of AZA against SA induced sub-chronic cardiotoxicity in rats through the evaluation of biochemical and histological parameters.

Thirty-two rats were apportioned into control, SA (10 mg kg^-1), SA (10 mg kg^-1) + AZA (25 mg kg^-1), and AZA (25 mg kg^-1) exposed group. Biochemical parameters were evaluated through different techniques including qRT-PCR, ELISA and already reported standard assays. Histological analysis was performed to validate the tissue damage. The curative potential of AZA was re-validated through in-silico approaches.

SA intoxication upregulated the expression of nuclear factor-kappa B (NF-κB), myeloid differentiation primary response 88 (MyD88), janus kinase 1 (JAK1), toll-like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), signal transducer and activator of transcription 3 (STAT3), interleukin-1β (IL-1β), interleukin-6 (IL-6), & cyclooxygenase-2 (COX-2) while increasing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Moreover, the enzymatic activities glutathione peroxidase (GPx), glutathione reductase (GSR), glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), heme-oxygenase-1 (HO-1) and glutathione (GSH) were lowered whereas the levels of Troponin I, ProBNP, C-reactive protein, LDH, troponin-T, BNP and CPK were exacerbated following the SA exposure. Furthermore, SA intoxication promoted the levels of Bax, Caspase-9, and Caspase-3 while lowering the levels of Bcl-2. Cardiac tissues showed impaired histology after SA administration. Nonetheless, AZA therapy excellently ameliorated cardiac toxicity owing to its anti-inflammatory, antioxidative and anti-apoptotic potential.

SA intoxication induced severe cardiac toxicity via disrupting biochemical and histological parameters while AZA excellently restored cardiac health profile. Collectively, AZA may serve as a cardioprotective agent in counteracting SA induced sub-chronic cardiotoxicity.