Impaired fracture repair is associated with sustained inflammation, mitochondrial dysfunction, and disrupted osteoblast–macrophage communication. Cuproptosis involves copper-dependent aggregation of lipoylated mitochondrial proteins and loss of iron–sulfur proteins, suggesting a potential connection between mitochondrial injury and osteoimmune dysfunction. This study investigated this relationship in an in vitro inflammatory–hypoxic model. Human hFOB 1.19 osteoblasts and THP-1-derived macrophages were studied under delayed-healing-like stress (DHS; 1
The expression and function of E3 ubiquitin ligase WWP1, as well as the post-translational modification of transcription factor KLF5, have rarely been systematically characterized in Crohn’s disease (CD). We established a TNBS-induced CD-like colitis mouse model and an LPS-stimulated intestinal epithelial cell model. Histopathological staining, Western blot, qPCR and immunofluorescence were used to detect the expression and distribution of WWP1, KLF5 and NF-κB pathway-related proteins in colon tissues. Co‑immunoprecipitation (Co‑IP) and ubiquitination‑related signal assays were performed to examine protein co‑precipitation and ubiquitin‑associated signals for KLF5. CCK-8, ELISA, qPCR and Western blot were applied to assess cell viability, inflammatory cytokine secretion and NF-κB activation. A KLF5 overexpression rescue experiment was conducted to indicate the regulatory relationship between WWP1 and KLF5. In colonic tissues of colitis mice, WWP1 expression was increased to 3.46-fold, while KLF5 expression was decreased to 0.47-fold compared with controls. The levels of pro-inflammatory TNF-α and IL-6 were elevated to 2.81-fold and 3.06-fold, accompanied by noticeable NF-κB pathway activation. Co‑IP assays detected both proteins within immunoprecipitated complexes. WWP1 overexpression was accompanied by altered KLF5 ubiquitination signals, changes in KLF5 protein abundance, and exacerbated LPS‑induced inflammation, whereas ectopic KLF5 overexpression partially mitigated this pro‑inflammatory phenotype. Immunofluorescence suggested the co-localization of WWP1 and KLF5 in colonic mucosal cells of mouse colon tissues. This study suggests that WWP1 may aggravate intestinal inflammation in CD‑like colitis through potential modulation of KLF5, accompanied by downstream changes in NF‑κB signaling activation. This work provides additional mechanistic insights into CD‑like colitis and identifies a potential mediator for further pre‑clinical exploration.
This study aimed to investigate the therapeutic effects of Zhixie Capsule (ZXC) on ionizing radiation (IR)-induced intestinal injury (radiation enteritis, RE) and to elucidate the underlying molecular mechanisms, focusing on the PI3K/AKT signaling pathway. A rat model of RE was established by 12 Gy abdominal IR. ZXC was administered daily for 7 days, and its effects were evaluated by monitoring body weight, disease activity index (DAI), colon length, histopathology (H E), and intestinal barrier integrity (Alcian blue staining, FITC-dextran permeability assay, and immunofluorescence for ZO-1 and occludin). Network pharmacology was used to predict key pathways. Inflammatory cytokines (IL-1β, IL-6, TNF-α, IL-10) were measured by ELISA, and pathway protein expression (p-PI3K, p-AKT, p-IKKβ, p-p65) was analyzed by Western blot. A rescue experiment using the PI3K activator YS49 was performed to confirm pathway specificity. In terms of results, ZXC dose-dependently alleviated RE manifestations, including weight loss, colonic shortening, and histological damage, and improved intestinal barrier integrity, as evidenced by increased goblet cell numbers and upregulated expression of tight junction proteins. Network pharmacology predicted the potential involvement of the PI3K/AKT axis. Mechanistically, ZXC suppressed IR-induced PI3K/AKT phosphorylation and downstream NF-κB activation, reducing pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and increasing IL-10 levels. These protective effects were partially reversed by YS49, suggesting that ZXC mitigates RE by inhibiting PI3K/AKT-mediated inflammation and improving mucosal barrier integrity, highlighting its potential as a radiomitigative strategy.
Obesity is a chronic metabolic disorder characterised by dyslipidemia, hyperglycemia, oxidative stress, inflammation and organ dysfunction. The anti-obesity drugs available are effective but adverse effects often limit their long-term use. Hydrocotyle sibthorpioides is a medicinal plant rich in flavonoid and phenolic compounds, with reported antioxidant and metabolic regulatory properties. The present study was aimed at investigating the anti-obesity potential of ethanolic extract of Hydrocotyle sibthorpioides (EEHS) and its phytosomal formulation in a high-fat diet (HFD)-induced obesity model. EEHS was characterised by phytochemical screening, thin-layer chromatography and determination of total phenolic and flavonoid contents. Obese male Wistar rats induced by HFD were treated with EEHS (100–400 mg/kg), EEHS phytosomes (400 mg/kg) or orlistat (25 mg/kg) for seven weeks. Physiological, biochemical, and histopathological parameters were assessed. EEHS exhibited significant anti-obesity effects in a dose-dependent manner, with the phytosomal formulation being the most significant efficacy. Treatment resulted in marked improvement in body weight, fasting blood glucose, lipid profile, liver enzyme levels, liver lipid accumulation, total body fat, TNF-α, antioxidant status and organ weights. Histopathological analysis confirmed reduced hepatic and vascular damage, inflammatory cell infiltration and adipocyte hypertrophy. The phytosomal formulation showed effects similar to those of orlistat and was more effective than the traditional extract. EEHS, especially as a phytosomal preparation, exhibited a potent anti-obesity effect. It ameliorated metabolic, inflammatory, oxidative, and histopathological alterations associated with HFD-induced obesity. The present findings suggest that EEHS may be a promising natural approach for the management of obesity.
Iron-dependent ferroptosis has emerged as an important pathological mechanism in acute lung injury, but the endogenous mechanisms that limit iron accumulation and lipid peroxidation during lung injury remain incompletely understood. We investigated whether DJ1 (Parkinson disease protein 7, encoded by PARK7), a cytoprotective protein induced by oxidative stress, limits iron accumulation and ferroptosis in lipopolysaccharide (LPS)-induced acute lung injury. Experiments were performed in male C57BL/6J mice challenged with intratracheal LPS and in Beas-2B human bronchial epithelial cells treated with the ferroptosis inducer erastin. LPS challenge in mice and erastin treatment in cells increased total iron content and produced changes consistent with ferroptosis, including glutathione depletion, malondialdehyde accumulation, and reduced expression of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11). C11-BODIPY analysis directly confirmed erastin-induced lipid peroxidation, and among the regulated cell death inhibitors tested, only ferrostatin-1 (Fer-1) substantially restored cell viability. DJ1 expression rose under both conditions. Adeno-associated virus-mediated DJ1 overexpression, delivered three weeks before LPS challenge, attenuated iron accumulation and lung injury, whereas DJ1 knockdown worsened both. Co-immunoprecipitation showed that DJ1 was associated with S-adenosylhomocysteine hydrolase (SAHH), an enzyme linked to methionine metabolism and glutathione synthesis. Higher DJ1 levels were accompanied by lower AHCYL1-SAHH association and higher SAHH activity. After treatment with adenosine periodate-oxidized (AdOx), the protection associated with DJ1 overexpression was no longer observed. Fer-1 produced protection similar to that associated with DJ1 overexpression. These findings support a role for DJ1 in limiting iron accumulation and ferroptosis-associated lung injury and are consistent with a contribution of SAHH activity. The precise contribution of the DJ1-SAHH-AHCYL1 relationship requires further mechanistic validation.
