We investigated how aging interacts with high-fat (HFD) and ketogenic (KD) diets to influence functional overload (FO)-induced muscle hypertrophy in C57BL/6J mice. Male mice aged 3, 16, and 24 months were fed regular chow (RD), HFD, or KD for 12 weeks, with FO of the plantaris muscle induced by denervation of the soleus and gastrocnemius during the final 6 weeks (n = 8-10 per group). Muscle hypertrophy was unaffected by diet (p = 0.897) but declined with age (p < 0.001), with a marked reduction already in mature mice. Plasma IGF-1 was higher in adult than mature (p < 0.05) and old (p < 0.001) mice and correlated positively with hypertrophy (r = 0.29, p = 0.01). The weak correlation of hypertrophy with plasma IGF-I levels suggests that IGF-I is only one among many factors playing a role in muscle hypertrophy.
Research increasingly implicates neuroimmune inflammation in schizophrenia, with the epidermal growth factor (EGF) family as a potential link. This review critically evaluates EGF family members as diagnostic, risk, and treatment response markers.The most consistent finding is decreased serum EGF in chronic schizophrenia, confirmed in postmortem brain and independent of antipsychotics. First-episode data are heterogeneous, reflecting differences in medication status, population, and illness stage.Genetic studies of EGF rs4444903 show no consistent disease risk association but demonstrate a replicable, sex-specific effect on age at onset in males. Stronger evidence implicates NRG1 type IV and NRG3 polymorphisms in disease biology.Serum EGF is not a reliable treatment response marker. In summary, EGF alone is not clinically useful. Its value lies in consistent reduction in chronic patients, sex-specific effects on age at onset, and robust links of NRG1 and NRG3 to schizophrenia.
ERK, a core member of the MAPK family, operates through the classical Ras/Raf/MEK/ERK signaling cascade. Abnormal activation of this pathway is closely associated with malignant tumor progression and represents a critical target in anticancer drug development. In this article, we systematically describe the regulatory network of the ERK pathway, including negative feedback mechanisms, cross-pathway interactions, non-coding RNAs, and epigenetic regulation, with a particular focus on ERK's dual role-promoting both cell proliferation and apoptosis-in pituitary adenomas compared to other tumors. Additionally, we explore emerging ERK-targeted therapies and their associated resistance mechanisms. Despite substantial research on ERK, further investigation into its underlying mechanisms is warranted. We propose the construction of computational modeling approaches to analyze these complex regulatory relationships and to develop precise combination therapies aimed at overcoming resistance. Such strategies could enable personalized treatment and offer more effective solutions for managing pituitary tumors and other malignancies.
Endometriosis, a chronic inflammatory condition linked to pelvic pain and infertility, is characterized by immune dysfunction involving dysregulated apoptosis and cell proliferation. This case-control study included 87 infertile women undergoing diagnostic laparoscopy and assessed serum and peritoneal levels of sEGFR, sFas, and sFasL using ELISA. Pain intensity was evaluated via the Visual Analog Scale (VAS). Peritoneal sFasL concentrations were significantly higher in infertile women with pelvic pain (p = 0.012) and correlated with VAS scores (r = 0.261, p = 0.014). Serum and peritoneal sFasL levels were significantly elevated in endometriosis patients with pelvic pain compared to controls (p = 0.003, p = 0.002, respectively). Peritoneal sFasL levels were also associated with dysmenorrhea and dyspareunia. ROC analysis demonstrated that serum sFasL differentiated endometriosis from controls (AUC = 0.797, p < 0.001). These findings suggest that sFasL may serve as a potential noninvasive biomarker for endometriosis. Further research should investigate its regulatory role in endometriosis-related pain.
The aim of the study was to evaluate the angiogenic potential and safety of Dabrafenib, a BRAF inhibitor, for wound healing applications using the chick embryo Chorioallantoic Membrane (CAM) assay. Fertilized chicken eggs were incubated, and CAM assay was performed to assess angiogenesis. The number of blood vessels was quantified using Image J software. An irritation test was conducted by applying Dabrafenib to the CAM and observing vascular changes. Additionally, the effects of dabrafenib on the secretion of Vascular Endothelial Growth Factor (VEGF-A) and Fibroblast Growth Factor 2 (FGF2) in HaCaT cell lines were investigated. Dabrafenib demonstrated a dose-dependent pro-angiogenic effect, with a EC50 of 1.196 mM. The irritation test indicated no significant irritation with Dabrafenib. There was a 4.2-fold and 5.6-fold increase from control in the VEGF-A and FGF2 groups, respectively. Dabrafenib demonstrated angiogenic potential and was nonirritating, indicating its safety for topical application.
Significant dysregulation of matrix metalloproteinase (MMPs), their tissue inhibitors (TIMPs) and profibrotic cytokines have been reported in hypertrophic cardiomyopathy (HCM) and associated with adverse cardiac remodeling. We aim to investigate the relationship among MMPs, TIMPs, transforming growth factor-β (TGF-β) and amino terminal propeptide of type III procollagen (PIIINP) in HCM patients, and to explore their associations with clinical outcomes. We recruited 46 HCM patients and 49 controls, and measured biomarkers levels using ELISA. Receiver operating characteristic (ROC) analysis revealed the highest diagnostic sensitivity for MMP-2 (93.3%) and specificity for TGF-β (90%). Significant correlations were found between biomarkers and clinical parameters, including interactions between TGF-β and left ventricular mass, MMP-2 and intraventricular septum, PIIINP and maximal wall thickness. We suggest that TGF-β, with the highest specificity and MMP-2, with high sensitivity, may offer significant potential for identifying patients with HCM and serve as useful biomarkers for this disease.
The majority of colon adenomas harbor genetic mutations in the APC gene. APC mutation leads to changes in Wnt signalling and cell-cell adhesion: as a consequence, intestinal crypt budding increases and the excess crypts accumulate to form adenomas that progress to colon cancer. When cultured with Wnt, R-spondin, EGF, Noggin, myofibroblast conditioned medium and Matrigel, crypts from normal mouse colon mucosa form crypt-producing organoids and can be passaged continuously. Under the same culture and passage conditions, crypts isolated from colon adenomas derived from Apcmin/+ mice typically grow as spheroidal cysts and do not produce crypts. The adenoma organoid growth requires EGF, but not Wnt, R-spondin or Noggin. However, when mouse colon adenoma spheroids are grown for more than 10 days in the presence of EGF, crypt formation occurs. EGF, EREG, β-cellulin, Neuregulin-1 or AREG are sufficient for initiating crypt formation, however, neuregulin-1 is more potent than the other EGF-family members. EGFR and ErbB2 inhibitors both prevent crypt formation in adenoma cultures. Either EGFR:ErbB2 or ErbB3:ErbB2 signalling is sufficient to initiate adenoma crypt budding and elongation. ErbB2 inhibitors may provide a therapeutic avenue for controlling and ablating colon adenomas.
Periodontitis causes severe tissue loss, making predictable regeneration a major challenge. Growth factor-loaded chitosan scaffolds offer a promising, multifunctional approach to enhance tissue regeneration. This review explores their design, optimization, and therapeutic potential, focusing on delivery methods, release kinetics, and biocompatibility. Studies show these scaffolds improve local delivery, sustain growth factor release, and reduce cytotoxicity while offering reinforcement and encouraging cell adhesion. Optimized release kinetics ensure a favorable prognosis, reducing the necessity for recurrent administration. Growth factor-loaded chitosan scaffolds create a biomimetic environment that supports periodontal healing, representing an innovative strategy for advancing periodontal therapy. Further research on scaffold design, growth factor combinations, and long-term outcomes is essential to refine this approach for more effective, predictable regeneration.
IGF1R, a receptor tyrosine kinase, is crucial for cell growth, proliferation, and differentiation. Overexpression of IGF1R is linked to cancer and resistance to therapies, making it a target for drug development. Previously, we identified an allosteric binding pocket in IGF1R, which could be exploited for the development of IGF1R specific small molecule inhibitors. A recent study linked M1054I variant to growth defects and microcephaly. In this study, we functionally characterized M1054I to assess its effects on kinase activity, downstream signaling, and receptor structure. Replacing methionine with isoleucine at position 1054 leads to complete loss of kinase activity. Consequently, activation of downstream signaling molecules is significantly reduced in cells carrying the M1054I mutation. Structural analysis reveals that the mutation causes conformational changes in key conserved regions, particularly in the activation loop, compromising IGF1R's structural integrity and function. Additionally, the roles of residues Y987 and K1033 in regulating IGF1R functions were examined.
myocardial ischemia-reperfusion injury (MI/RI) threatens people's lives. Midkine (MDK) has a protective effect against MI/RI. DNA methyltransferase 1 (DNMT1) is closely associated with MI/RI. Mouse cardiomyocytes were isolated to establish a model of myocardial ischemia-reperfusion. Subsequently, the viability, apoptosis, and inflammatory factors of the cardiomyocytes treated under different conditions were measured. Western blotting was employed to examine the expression of related proteins. Chromatin immunoprecipitation (CHIP) and dual-luciferase reporter gene assays were conducted to verify the relationships among the DNMT1, MDK, and Notch2/Hes1 signaling pathways. The expression of MDK was reduced in the MI/RI cell model, while Notch2/Hes1 was activated. Overexpression of MDK mitigated the injury associated with MI/RI. Methylation modification of DNMT1 downregulated MDK, thereby mediating the Notch2/Hes1 pathway. The methylation modification of DNMT1 on MDK regulated the progression of MI/RI. The methylation modification of MDK by DNMT1 likely downregulates the Notch2/Hes1 pathway, thereby influencing MI/RI.
During the formation of hypertrophic scars (HS), there is often a notable abnormal proliferation and differentiation of cells, especially fibroblasts, but it remains ambiguous whether a causal relationship exists between fibroblast growth factor receptors (FGFRs) and hypertrophic scar. This study explored the causal impact of FGFR1, FGFR2, FGFR3, and FGFR4 on HS utilizing a two-sample Mendelian randomization (MR) analyses. The elevated expression of FGFR1 and FGFR 4 emerged as two potential risk factors against HS in the inverse-variance weighted analysis. Conversely, FGFR2 and FGFR3 exhibited no significant causal relationship with hypertrophic scars. Rigorous analyses including assessments of heterogeneity, genetic horizontal pleiotropy, and leave-one-out sensitivity collectively affirmed the stability and reliability of the findings in this study. Taken together, the elevated expression of FGFR1 and FGFR 4 act as two key regulatory factors in preventing the formation of HS and serves as a crucial modulator in impeding scar formation.
GATA proteins have been proposed as regulators of adipogenesis. In particular, GATA2 and GATA3 have been demonstrated to function in preadipocytes, and their behavior in response to extracellular signals allows preadipocytes to transition into adipocytes in mammals. However, few reports have discussed the extracellular stimulators of both GATAs. In this study, we investigated whether growth hormone (GH) acts as an extracellular ligand of both GATAs, with special attention paid to GATA3 expression in adipocytes. GATA3 expression increased at the transcriptional level with STAT5B activation in a dose-dependent manner, and this was reversed by a STAT5-specific inhibitor. Furthermore, we found that the functional STAT5B consensus sequences that function as the binding site in the GATA3 promoter region were located at -117 bp from the transcription starting site. These results suggest that GH induces GATA3 gene expressions via the GHR/JAK/STAT5 pathway to control adipocyte proliferation and differentiation.
Optic nerve (ON) injury leads to retinal ganglion cell (RGC) degeneration and axonal atrophy. Wnt ligands are embryonic growth factors that regulate cellular differentiation and survival. We recently demonstrated that canonical and non-canonical Wnt signaling induces RGC survival and axonal regrowth after optic nerve crush (ONC) injury in mouse. Here, we investigated whether the non-canonical Wnt5a ligand induces pro-regenerative inflammation after ONC. Mice were intravitreally injected with Wnt5a or saline during ONC and retina tissue was collected for QPCR and immunofluorescence. We demonstrated that expression of arginase 1, a marker of anti-inflammatory microglia, was upregulated by Wnt5a in injured retinas, whereas iNOS, a marker of neurotoxic microglia, was suppressed. Wnt5a also induced time-dependent changes in pro-inflammatory genes Gal3, TNFα, P2RY12 and IL-6 and the anti-inflammatory gene IL-27. These results indicate that Wnt5a is an immunomodulatory ligand in the retina after ONC injury.
Vascular endothelial growth factor (VEGF) plays a crucial role in maintaining renal homeostasis. However, the precise impact of VEGF on ferroptosis in acute kidney injury (AKI) remains incompletely understood. This study aims to investigate the effects of VEGF on ferroptosis in the model of AKI and to elucidate the underlying mechanisms. We used C57BL mice and HK-2 cells to construct sepsis-associated AKI models. We assessed renal function, cell viability, and tissue levels of iron, malondialdehyde (MDA), and glutathione (GSH) in mice. Intracellular reactive oxygen species (ROS), mitochondrial membrane potential, and electron microscopy-detected cellular changes were also measured. Western blotting analyzed key ferroptosis-related proteins (SLC7A11, GPX4) and components of the ERK1/2-NRF2-GPX4 pathway. VEGF treatment significantly reduced oxidative stress by lowering ROS and MDA levels while increasing GSH. Additionally, VEGF165 activated the ERK1/2-NRF2 pathway, mitigating ferroptosis.
To identify distinct patterns of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-binding protein 3 (IGFBP-3) in girls with central precocious puberty (CPP), and to compare IGF-1 and IGFBP-3 levels in patients with CPP and precocious thelarche (PT). A literature search of PubMed, Embase, and Scopus databases was done to identify observational studies. Newcastle-Ottawa Scale (NOS) was used to assess the quality of the studies. Pooled weighted mean difference (WMD) and 95% confidence intervals (CI) were reported. Fourteen studies were included. As opposed to age-matched or similar-aged pre-pubertal girls, patients with CPP had significantly higher levels of IGF-1 and IGFBP-3. CPP was associated with higher levels of IGF-1 approaching statistical significance but similar levels of IGFBP-3 compared to PT. Distinct hormonal patterns, such as elevated IGF-1 and IGFBP-3, were identified in patients with CPP, suggesting a potential diagnostic value of IGF-1 and IGFBP-3 levels.
Systemic inflammatory response syndrome (SIRS) commonly considered as the first step in the sepsis cascade. To evaluate the diagnostic value and potential mechanism of serum MANF in sepsis-associated lung injury (SALI), 15 adult SALI patients and 15 age- and sex-matched SIRS patients were enrolled to measure serum MANF levels by sandwich enzyme-linked immune-sorbent assay and the laboratory indexes including C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) were collected. MANF and IL-6 levels showed significant differences between groups (P < 0.05), and the area under the curve (AUC) values of MANF and IL-6 were 0.822 and 0.815. The combination of MANF with IL-6 exhibited improved predictive accuracy for SALI. Furthermore, we found that there is a protein interaction network between MANF and all of these overlap targets between ferroptosis-related genes (FRGs) and SALI by bioinformatics analysis. MANF could be a promising biomarker for the differential diagnosis of SALI and SIRS.
Anti-angiogenics, inhibitors of pathological blood vessel growth, are an important class of targeted agent for the treatment of common cancers and ocular conditions. However, efficacy is compromised by the absence of biomarkers to guide patient selection or inform the management of resistance. We describe an assay for modified endothelial cell (EC) responses to the VEGF-A-neutralizing monoclonal antibody bevacizumab as part of a biomarker discovery program. ECs are transduced by lentivector expressing an experimental or non-silencing shRNA, each co-expressed with a different fluorescent protein. A 1:1 mixed cell population is then cultured with bevacizumab or control antibody under VEGF-A-dependent conditions. A normalized ratio of surviving cells, obtained by flow cytometry analysis, reflects EC resistance or sensitization to bevacizumab mediated by the experimental shRNA. With reagents prepared, the protocol takes 10 days and rigorously quantifies the impact of gene perturbation on the EC response to bevacizumab or other targeted anti-angiogenics.
Pancreatic endocrine cells are categorized in to 5 types (alpha, beta, delta, pancreatic polypeptide cells and epsilon), which expresses glucagon, insulin, somatostatin, pancreatic polypeptide, and ghrelin, respectively. Several studies including lineage tracing in Ins2Akita diabetic mice have been done to investigate the identities of pancreatic endocrine cells which concludes, alpha cells have enormous plasticity, which enables them to be reprogrammed by specific transcription factors into insulin secreting beta like cells. Gene therapy has provided the beneficial outcome. Pdx1, MaFA and PAX4 (the transcription factors) in alpha cells can be over expressed which results in reprogramming the targeted alpha cells into beta cells. This trans-differentiation may be induced by infusing an adeno-associated virus (AAV) loaded with distinct transcription factors in the duct of pancreas. Several researches have demonstrated the successful restoration of enhanced insulin secretion in diabetes induced mice. Additionally ductal neurogenin3 (Ngn3), Sglt2 inhibitors, Igfbp1, GLP1 and several clinical and non-clinical agents has been postulated as a basis of beta cell neogenesis. Alpha cell owing to its high plasticity, on prolonged exposure to GABA reprogrammed into beta-like cell due to downregulation of Arx expression by GABA. The various approaches for beta cell neogenesis open a new window towards the establishment of novel gene therapy accession to treat diabetes. However, broad studies are still needed to improve and optimize this treatment methodology. The potentiality of endogenous pancreatic alpha cell to beta cell conversion methods and its outcomes are invigorating. This accomplishment is presently being under trial in non-human primates.
Oxaloacetate (OA) is a pivotal endogenous metabolite. Within our investigation, we ascertained that OA functions as an agonist for the epidermal growth factor receptor (EGFR), a key protagonist in the genesis of diverse tumours. We substantiated that escalating concentrations of OA initially enhanced the cellular viability of several cancer cells, followed by subsequent attenuation, which is similar to the effect of EGF. Furthermore, the protein phosphorylation profile in HepG2 cells exposed to OA closely paralleled that induced by epidermal growth factor (EGF). Additional findings underscored the capability of OA to induce the generation of EGFR dimers. Finally, our observations revealed that OA governs the activation of AKT and Erk, the typical downstream signalling proteins of EGFR. We postulate that the endogenous metabolite OA can function as either an agonist or inhibitor of EGFR at specific concentrations to modulate tumour proliferation, and provide new insights into the regulation of EGFR activation.
Activated Akt and loss of phosphatase and tensin homolog (PTEN) tumour suppression aid chemo- and radio-resistance in glioblastoma stem cells (GSC), contributing to treatment failure in glioblastoma. In this study, sixteen GSC lines were generated from 66 individual glioma samples, in gliomasphere culture conditions. Thirteen of 16 GSC lines expressed hyperphosphorylated Akt (Ser473); Akt phosphorylation did not correlated with EGFR expression. An LDH colorimetric assay was used to measure the in vitro cytotoxicity of eight of these lines. Akt X (20 µM) proved more effective at inducing in vitro GSC cytotoxicity (range: 22-73%) over 48 hours than triciribine (20 µM) (0-27%), although both agents inhibited Akt phosphorylation as detected by western blot analysis. A statistically significant correlation between PTEN loss (western blot) and the extent of Akt X-induced cytotoxicity was found (p = 0.03). Akt inhibition reduces in vitro proliferation of treatment-resistant GSC lines, especially in PTEN-deficient lines, warranting further translational investigation in glioblastoma.
