TRPC1 channel modulates mechanical stretch-induced bone marrow mesenchymal stem cell proliferation through Ca2+-dependent ERK1/2 activation.

A stimulating effect induced by Bisphenol A bis (diphenylphosphate) in HepG2 cells via vascular endothelial growth factor (VEGF)/VEGF receptor axis.

S-nitrosylation of Dexras1 attenuates fear memory generalization in the infralimbic cortex.

Delivery growth factors by layer-by-layer assembly in nanofibers for enhancing bone defect repairment along with neurogenesis.

Persistent facial and oral discomfort, particularly trigeminal neuralgia (TN), is frequently accompanied by anxiety, which has been closely linked to increased excitability of neurons in the lateral habenula (LHb). However, the mechanisms underlying this hyperexcitability remain unclear. Here, we show that partial transection of the infraorbital nerve (pT-ION) significantly upregulated the expression of transient receptor potential canonical 6 (TRPC6), β isoform of calcium/calmodulin-dependent protein kinase II (βCaMKII), phosphorylated extracellular regulated kinase (p-ERK), and phosphorylated cyclic adenosine monophosphate response element-binding protein (p-CREB) in the LHb. Pharmacological blockade of either TRPC6 or βCaMKII effectively reversed pT-ION-induced mechanical hypersensitivity and anxiety-like behaviors. TRPC6 overexpression in the LHb reproduced the behavioral and electrophysiological phenotypes observed in pT-ION mice, including increased LHb neuronal excitability. In contrast, bilateral knockdown of TRPC6 attenuated both pain- and anxiety-like behaviors and normalized neuronal activity in the LHb. Our study identified TRPC6 as a key mediator of LHb neuronal hyperexcitability, contributing to trigeminal neuralgia-associated pain and anxiety via the βCaMKII/ERK/CREB pathway, and suggests its potential as a target for treatment.

Objective: To investigate the role of Rap1GAP in myocardial hypertrophy and fibrosis and its potential mechanism. Methods: In animal experiments, 20 cardiac-specific Rap1GAP knockout mice (Rap1GAPΔ/Δ) and 20 Rap1GAP homozygous floxed mice (Rap1GAPfl/fl), aged 6-8 weeks, were divided into four groups: Rap1GAPfl/fl+saline group, Rap1GAPΔ/Δ+saline group, Rap1GAPfl/fl+AngⅡ group, and Rap1GAPΔ/Δ+AngⅡ group. Echocardiography was used to assess cardiac function in each group. HE staining was performed to observe the overall structure and cellular morphology of myocardial tissue. Wheat germ agglutinin staining was used to measure the cross-sectional area of cardiomyocytes. Masson staining and Sirius red staining were employed for quantitative analysis of myocardial fibrosis levels and collagen proportion. Western blot was used to detect the protein expression levels of Rap1GAP, α-smooth muscle actin (α-SMA), atrial natriuretic peptide (ANP), β-myosin heavy chain (β-MHC), collagen type Ⅰ, collagen type Ⅲ, transforming growth factor-β1 (TGF-β1), phosphorylated extracellular regulated kinase (p-ERK), extracellular regulated kinase (ERK), phosphorylated c-Jun N-terminal kinase (p-JNK), c-Jun N-terminal kinase (JNK), phosphorylated p38 mitogen-activated protein kinase (p-p38), p38 mitogen-activated protein kinase (p38), phosphorylated nuclear factor κB (p-NF-κB), and nuclear factor κB (NF-κB) in myocardial tissue. In cell experiments, primary cardiomyocytes and cardiac fibroblasts were isolated from 1-3-day-old Wistar rat neonates. Rap1GAP expression was knocked down by transfecting Rap1GAP small interfering RNA (si-Rap1GAP), and cells were divided into si-control (negative control siRNA)+saline group, si-Rap1GAP+saline group, si-control+AngⅡ group, and si-Rap1GAP+AngⅡ group. Rap1GAP was overexpressed by infecting with Rap1GAP adenovirus (Ad-Rap1GAP), and cells were divided into Ad-GFP (GFP empty vector adenovirus)+saline group, Ad-Rap1GAP+saline group, Ad-GFP+AngⅡ group, and Ad-Rap1GAP+AngⅡ group. For inhibitor rescue experiments, cardiac fibroblasts infected with Ad-Rap1GAP or Ad-GFP were treated with 10 μmol/L p38 inhibitor (SB203580) or TGF-β1 inhibitor (pirfenidone), forming Ad-Rap1GAP+AngⅡ+SB203580 group, Ad-Rap1GAP+Ang+pirfenidone group, Ad-GFP+AngⅡ+SB203580 group and Ad-GFP+Ang+pirfenidone group. Immunofluorescence staining was used to detect the expression levels of Rap1GAP, α-SMA, and proliferating cell nuclear antigen (PCNA) in cells. Dihydroethidium staining was employed to measure reactive oxygen species (ROS) levels. Western blot was used to detect the expression levels of target proteins (consistent with animal experiments). Results: In animal experiments, compared with the Rap1GAPfl/fl+AngⅡ group, the Rap1GAPΔ/Δ+AngⅡ group showed larger left ventricular end-diastolic diameter and left ventricular end-systolic diameter, while the cardiomyocyte surface area, myocardial fibrosis ratio and myocardial collagen volume ratio were smaller. Additionally, the expression levels of ANP, β-MHC, collagen type Ⅰ, collagen type Ⅲ, and α-SMA were lower (all P<0.05). Proteomic analysis revealed differences in the protein expression profiles of myocardial tissue between the Rap1GAPΔ/Δ+AngⅡ group and the Rap1GAPfl/fl+AngⅡ group at biological process, cellular component, and molecular function. In cell experiments, compared with the si-Rap1GAP+AngⅡ group, the si-control+AngⅡ group exhibited larger cardiomyocyte cross-sectional area, as well as higher expression levels of ANP, β-MHC, collagen type Ⅰ, collagen type Ⅲ and α-SMA, and a higher proportion of PCNA-positive cells (all P<0.05). Compared with the Ad-GFP+AngⅡ group, the Ad-Rap1GAP+AngⅡ group showed larger cardiomyocyte cross-sectional area, higher expression levels of the aforementioned proteins, and a higher proportion of PCNA-positive cells (all P<0.05). Furthermore, the relative expression levels of p-ERK/ERK, p-JNK/JNK, p-p38/p38, p-NF-κB/NF-κB proteins, and ROS levels in cardiomyocytes or cardiac fibroblasts were higher in the si-control+AngⅡ group than in the si-Rap1GAP+AngⅡ group (all P<0.05), while these indices were higher in the Ad-Rap1GAP+AngⅡ group than in the Ad-GFP+AngⅡ group (all P<0.05). Meanwhile, the ROS level, proportion of PCNA-positive cells, and expression levels of TGF-β1, p-ERK/ERK, p-JNK/JNK, p-p38/p38, collagen type Ⅰ, collagen type Ⅲ, and α-SMA proteins in cardiac fibroblasts were lower in the Ad-GFP+Ang+SB203580 group than in the Ad-GFP+AngⅡ group (all P<0.05). The same trend was observed in the Ad-GFP+AngⅡ+pirfenidone group, with all aforementioned indices lower than those in the Ad-GFP+AngⅡ group (all P<0.05). Conclusions: Rap1GAP may mediate cardiomyocyte hypertrophy by regulating the mitogen-activated protein kinase signaling pathway and the expression of its downstream target NF-κB. Meanwhile, Rap1GAP may promote myocardial fibrosis by inducing ROS production and activating the TGF-β1/mitogen-activated protein kinase signaling pathway.

Extracellular regulated kinase (ERK) signaling is a major driver of cancer development. Mutations accumulated in oncogenes upstream of ERK can promote and sustain tumorigenesis by providing a sustained proliferative signal, helping cells resist death, and inducing angiogenesis. Therapeutic strategies for cancers with dysregulated ERK signaling have focused on inhibiting upstream-mutated oncogenes as a means of depriving tumors of these essential survival cues. While these strategies have demonstrated initial clinical success in patients, they also represent an incomplete understanding of the more nuanced role ERK signaling plays in tumor cell homeostasis. It is now understood that increased ERK signaling can also function as a tumor suppressor by inducing proliferative arrest outside of the framework of oncogene-induced senescence. In this review, we highlight current research describing the vulnerability of cancer cells to ERK hyperactivation induced toxicity and offer insight on how ERK rewiring may be leveraged for the development of new therapeutic strategies for patients.

Phosphatidic acid (PA) is increasingly recognized as an important endogenous regulator of cell proliferation and migration, playing relevant roles in physiology and pathology. However, the potential and prominence of extracellular PA in controlling cell functions are not so well established. The present review article has been undertaken to update and discuss the latest findings on extracellular PA as regulator of cell homeostasis, with special attention being paid to its role in the regulation of cell growth and migration. Specifically, exogenous PA potently stimulates myoblast proliferation and lung cancer cell migration, pointing to a critical role of this glycerophospholipid in the regulation of muscle cell regeneration and lung cancer dissemination. Interestingly, both of these actions are mediated through interaction of PA with lysophosphatidic acid (LPA) receptors and the subsequent activation of different signal transduction pathways. In particular, PA induces mitogen-activated protein kinase kinase (MEK)/extracellularly regulated kinases (ERK) 1 and 2, phosphatidylinositol 3-kinase (PI3K)/Akt, focal adhesion kinase (FAK)/Rac1, and Janus kinase-2 (JAK2)/signal transducer and activator of transcription 3 (STAT3). These findings may contribute to a better understanding of muscle cell biology and may help to develop new therapeutic strategies to treat lung cancer dissemination.

To analyze the functional impact of a rare heterozygous variant of SOS1 gene (c.283G>A, p.E95K) identified in a fetus with cervical cystic hygroma and to explore its association with the disease phenotype. A pedigree analysis was carried out to evaluate the co-segregation of the variant with the disease phenotype. Bioinformatic tools were employed to assess the conservation, protein structure and stability. Functional validation was conducted on HEK293T cells using fluorescence quantitative reverse transcription-PCR and Western blotting to measure the expression of SOS1 and phosphorylation levels of extracellular regulated protein kinases (ERK) and c-Jun N-terminal kinase. A literature review of previously reported disease-associated SOS1 variants was also carried out. This study has been approved by the Medical Ethics Committee of West China Second University Hospital, Sichuan University (Ethics No.: 201940). The variant was inherited from the husband of the woman with distinctive facial features and has co-segregated with the phenotype. Bioinformatics analysis indicated that the variant is located in a highly conserved region, and that p.E95K could disrupt key amino acid interactions and protein stability. Multiple bioinformatic predictions consistently suggested the pathogenicity of this variant. Functional assays demonstrated reduced SOS1 protein expression and decreased ERK phosphorylation. This study has revealed the functional impact of the SOS1 c.283G>A (p.E95K) variant, suggesting that it may contribute to the developmental phenotypes through a haploinsufficiency mechanism.

The ethanolic extract of the roots of Murraya exotica (Rutaceae) yielded twenty coumarins, including twelve previously undescribed compounds named exoticoumarins A-L (1-12; two biscoumarins, five coumarin hybrids, and five monomers). Their structures, including absolute configurations, were elucidated by a combination of NMR and HR-ESI-MS analyses, single-crystal X-ray diffraction, ECD exciton coupling, Mo2(OAc)4- and Rh2(OCOCF3)4-induced ECD, comparison of experimental with calculated ECD spectra, and chemical hydrolysis. Anti-inflammatory evaluation in LPS-stimulated RAW264.7 macrophages identified exoticoumarins A and K (1 and 11) as potent inhibitors of nitric oxide (NO) production, with IC50 values of 7.41 and 10.63 μM, respectively. Mechanistic studies revealed that 1 suppressed nitric oxide synthase (iNOS) expression at both transcriptional and translational levels, an effect associated with the inhibition of c-Jun N-terminal kinase (JNK) phosphorylation within the mitogen-activated protein kinase (MAPK) signaling pathways, without markedly affecting extracellular regulated protein kinases (ERK) 1/2 phosphorylation. These findings highlight exoticoumarin A (1) as a promising anti-inflammatory lead derived from M. exotica.

This article is being retracted from Bioscience Reports at the request of the Editor-in-Chief and the Editorial Board. This follows the receipt of a notification from a reader, alerting the Editorial Board to irregularities in the flow cytometry graphs, some of which suggest that the graphs could have been hand-drawn. The authors were contacted regarding the concerns and the retraction but have not yet responded or provided requested raw data. Given the extent of the issues raised, the Editorial Board stand by the decision to retract the article.

Cognitive impairment (CI) is prevalent in clinical patients with heart failure (HF). However, there are no effective pharmacological interventions for CI treatment. Astragaloside IV (AS-IV) is the principal constituent of Astragalus Radix and effective in HF, but its action on CI after HF has not yet been studied. Cognitive function was evaluated in behavioural experiments on male rats with HF. Transcriptomics and network pharmacology were used to identify potential pathways. Western blots, quantitative real-time polymerase chain reaction (qRT-PCR), Elisa, and electron microscopy were used to validate changes in identified pathways, brain inflammatory cytokines, postsynaptic membrane glutamate receptors and levels of apoptosis. Molecular docking and molecular dynamics simulation confirmed the binding ability of AS-IV and selected core proteins. AS-IV improved cardiac function and disordered cardiac muscle structure in rats with HF. The advanced glycation end-products-receptor of advanced glycation end-products (AGE-RAGE) signalling pathway was the core pathway. AS-IV reduced the expression levels of RAGE, extracellular regulated protein kinases 1/2 (ERK1/2) and nuclear factor kappa-B (NF-κB). AS-IV also attenuated the expression of proinflammatory factors, increased the levels of anti-inflammatory factors, and up-regulated postsynaptic membrane glutamate receptor expression. Molecular docking and molecular dynamics simulation showed that AS-IV had favourable binding energy with RAGE. AS-IV enhances cognitive function of HF rats by inhibiting the RAGE-ERK1/2-NF-κB signalling pathway to reduce inflammation and apoptosis and thereby improve synaptic function. AS-IV emerges as a promising candidate for the prevention and treatment of HF-CI.

Endometrial epithelial cell proliferation is essential for establishing endometrial receptivity during embryo implantation. In ruminants, embryonic interferon-tau (IFNT) mediates receptivity establishment by activating endometrial interferon-stimulated genes. This study investigates how IFNT-induced interferon alpha-inducible protein 6 (IFI6) regulates bovine endometrial epithelial cells (bEECs) proliferation. We identified the JAK2-STAT3 (rather than STAT1) as the primary signaling axis driving IFNT-mediated IFI6 expression. Functional experiments revealed that IFI6 knockdown markedly impaired bEECs proliferation and suppressed extracellular-regulated protein kinase (ERK1/2) phosphorylation, while IFNT supplementation reversed these defects. Notably, pharmacological activation using RO8191 (an IFN receptor agonist) restored STAT1/3 signaling and ERK1/2 activation in IFI6-knockdown bEECs, normalizing their proliferation rates. Mechanistically, IFI6 functions upstream of c-Jun/c-Fos, the major functional form of activating protein-1 (AP-1), and its knockdown disrupted ERK1/2-dependent regulation by preventing c-Jun/c-Fos heterodimerization and nuclear translocation. Our findings reveal that IFI6 maintains proliferative balance in bEECs by functioning as a positive feedback modulator of the IFNT-activated Janus kinase signal transducer and activator of transcription (JAK-STAT) pathway, which subsequently triggers the ERK1/2-c-Jun/c-Fos signaling axis. These results provide novel insights into IFNT-mediated receptivity regulation and highlight IFI6 as a potential diagnostic marker for early pregnancy loss, as well as a therapeutic target for enhancing implantation success.

We investigated the effects of hydrogen sulfide (H2S) and sonic hedgehog (SHH) on the proliferation, autophagy, and apoptosis of human microvascular endothelial cells (HCMEC/D3). We also explored the regulatory relationship between cystathionine-β-synthase (CBS) and the SHH pathway. Human microglia cells (HMC3) were stimulated under hypoxia to secrete H2S and SHH proteins, which were then co-cultured with HCMEC/D3 cells. The relationship between H2S and SHH was investigated by inhibiting the CBS or SHH pathways. Vascular endothelial growth factor (VEGF) and H2S levels were detected using ELISA. The mRNA and Protein levels of VEGF, Beclin-1, light chain 3 (LC3), Cysteine aspartic acid protease-3(caspase-3), CBS, SHH, extracellular regulated kinase 1/2 (ERK1/2) and phospho-ERK1/2 (P-ERK1/2) were determined by RT-PCR and western blot. The results indicated that H2S secretion by HMC3 increased during hypoxia, with both CBS and SHH proteins being up-regulated. The inhibition of CBS resulted in decreased levels of H2S and SHH in HMC3. When the SHH pathway is inhibited, H2S secretion levels remain unaffected. H2S and SHH proteins increased VEGF, P-ERK1/2, Beclin-1, and LC3 expression while reducing caspase-3 expression in HCMEC/D3 cells. H2S secretion by HMC3 promotes the proliferation and regeneration of HCMEC/D3 by regulating SHH protein and alleviating hypoxic injury.

Plasma accumulation of the gut microbial metabolite 4-ethylphenylsulfate (4EPS), derived from dietary amino acid, tyrosine, has been associated with cardiovascular, renal, metabolic, and neurological disorders. AngII (angiotensin II) infusion increases circulating 4EPS in mice, suggesting a potential mechanistic role. We hypothesized that 4EPS modulates AngII-regulated pathophysiology and disease progression by directly inhibiting AT1R (angiotensin II type 1 receptor). This hypothesis was tested by combining AT1R pharmacology, cell signaling assays, ex vivo vascular studies, an AngII-induced aortic aneurysm growth model, and plasma proteomics analysis. in vitro, 4EPS reduced the binding of both AngII and the antagonist candesartan to AT1R and suppressed AngII-induced calcium signaling. Ex vivo, 4EPS attenuated AngII-mediated vasoconstriction. In vivo, high-fat diet-fed ApoE-null mice coinfused with AngII and 4EPS showed significant blunting of blood pressure elevation and a marked reduction in aortic aneurysm-related mortality compared with mice infused with AngII alone. Analysis of aortic remodeling revealed increased elastin preservation and decreased thickening of the intimal and medial layers in 4EPS-treated animals. Plasma proteomics indicated alterations in actin-cytoskeletal signaling pathways consistent with reduced activation of ERK (extracellular-regulated kinase) 1/2, filamin-A, and proteins involved in vascular smooth muscle cell motility. These findings identify 4EPS as a benign, endogenous AT1R antagonist that diminishes AngII-mediated hemodynamic and vascular pathology. By suppressing cytoskeletal signaling associated with vascular remodeling, 4EPS provides significant protection against hypertension and aortic aneurysm progression in mice, revealing a previously unrecognized protective role for a gut microbial metabolite in modulating renin-angiotensin system activity.

Poly(ADP-ribose) polymerase (PARP) inhibitors (PARPis) are an important therapy for high-grade serous ovarian cancer (HGSOC). However, PARPi resistance frequently emerges, necessitating previously unrecognized approaches to improve HGSOC responses. Here, we showed that the anaplastic lymphoma kinase (ALK) inhibitor brigatinib enhances PARPi activity in HGSOC cells by disrupting an adaptive survival mechanism orchestrated by Fos-related antigen 1 (FRA1) in response to PARPi. This effect of brigatinib occurred through an ALK-independent pathway, wherein brigatinib induced a dual blockade of focal adhesion kinase (FAK) and EPH receptor A2 (EPHA2) tyrosine kinases, leading to the suppression of protein kinase B (Akt) and extracellular-regulated kinase (ERK) signaling accompanied by disruption of a phosphorylation event crucial for FRA1 protein stability. Moreover, in HGSOC patient-derived xenograft (PDX) models, brigatinib and PARPi combination therapy induced tumor regression and improved overall survival compared with PARPi alone, particularly in models with high FAK and EPHA2. These findings support dual targeting of FAK and EPHA2 as a strategy to achieve effective and durable PARPi responses and identify a promising biomarker-based combinatorial approach using brigatinib and PARPi for HGSOC, particularly the subset characterized by high FAK and EPHA2.

The efficacy of paclitaxel (PTX), an important chemotherapy drug in non-small-cell lung cancer (NSCLC) cells, is limited by its resistance. APG-1252 has an inhibitory role in cancer and can inhibit myeloid cell leukemia 1 (MCL-1) to enhance chemotherapy drugs' antitumor effect. Therefore, this research further investigated whether APG-1252 could enhance the anticancer role of PTX by regulating MCL-1. NSCLC cells were treated with drugs. Cell viability was determined using the cell counting kit-8 assay. The cell apoptosis was examined using flow cytometry and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling staining. The abilities of cells to migrate, invade, and proliferate were examined using transwell and colony formation assays, as needed. The expression levels of extracellular regulated protein kinases (ERK), phosphorylated ERK (p-ERK), and MCL-1 were quantified by Western blot. APG-1252 and PTX decreased the NSCLC cells' viability. APG-1252 and PTX induced cell apoptosis and inhibited the cells from migrating, proliferating, and invading. APG-1252 and PTX suppressed the expression of p-ERK and MCL-1. Besides, APG-1252 enhanced the anticancer role of PTX in NSCLC cells. Ro 67-7476, a p-ERK agonist, had an opposite role to the combination of APG-1252 with PTX in NSCLC cells. Additionally, Ro 67-7476 abolished the anticancer role of the combination of APG-1252 with PTX in the biological functions of NSCLC cells. APG-1252 enhanced the anticancer role of PTX in NSCLC cells by suppressing the ERK/MCL-1 pathway. This work provided the theoretical basis for the APG-1252 application.

Aberrant phosphorylation of receptor tyrosine kinases (RTKs) is commonly associated with tumorigenesis and metastasis. However, developing specific molecular tools to regulate RTKs phosphorylation and downstream signaling remains challenging. Herein, we develop a programmable and modular membrane-anchored DNA artificial receptor that enables the spatioselective modulation of RTK-related cellular signaling. As a proof of concept, the designed artificial receptor is anchored on a cell membrane with high stability to regulate and monitor the dimerization state of a typical RTK, fibroblast growth factor receptor (FGFR). Real-time observation of the dynamic regulation process is achieved based on single-particle tracking, providing an in-depth perspective of the modulation mechanism from the single-receptor level. Additionally, the artificial receptor system can effectively inhibit the activation of FGFR signaling even in the presence of natural ligands, which reduces protein kinase B and extracellular-regulated kinase 1/2 phosphorylation, further resulting in the cytoskeletal reorganization and upregulation of the pro-apoptotic protein expression. The proposed artificial receptor provides a novel chemical tool to regulate and visualize RTK-associated signaling pathways with tailored stoichiometry and spatial control, providing deep insights into the systematic investigation of receptor-mediated cellular signaling.

To explore the antioxidative effects of gypenosides (Gyps) in orbital fibroblasts (OFs) derived from patients with thyroid-associated ophthalmopathy (TAO) and to validate their therapeutic efficacy in vivo using an animal model. Bioinformatics analyses were performed to screen potential genes and signalling pathways underlying the effect of Gyps in OFs. OFs were isolated from orbital connective tissues of both TAO patients and non-TAO controls. CCK-8 assay was used to detect cell proliferation. Oxidative stress was evaluated by measuring reactive oxygen species (ROS) and superoxide dismutase (SOD) levels. Quantitative reverse transcriptive-polymerase chain reaction (RT-qPCR), ELISA, and western blotting were employed to ascertain the effects of Gyps on H2O2-induced oxidative stress, inflammation, fibrosis, and autophagy. Furthermore, a TAO mouse model was established. Gyps were administered by intraperitoneal injection (50 mg/kg/day for 4 weeks) to evaluate their protective effects against oxidative stress, inflammation, and fibrosis in orbital tissues. Bioinformatic analysis revealed that the identified genes were primarily enriched in metabolic and oxidative stress-related pathways. In vitro experiments demonstrated that Gyps significantly reduced H2O2-induced ROS generation, increased SOD levels, and suppressed the expression of inflammation-, fibrosis-, and autophagy-related markers. These effects were associated with the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2)/extracellular-regulated kinase (ERK)/heme oxygenase 1 (HO-1) pathway. In vivo animal experiments further confirmed that Gyps treatment effectively alleviated oxidative injury, inflammatory cell infiltration, and collagen deposition in the orbital tissues of TAO model mice. Gyps exert significant antioxidative, anti-inflammatory, and anti-fibrotic effects by activating the Nrf2/ERK/HO-1 signalling pathway in both in vitro and in vivo TAO models.

Histamine induces vascular endothelial cell proliferation via the histamine H1 receptor-extracellular regulated protein kinase 1/2-cyclin D1/cyclin-dependent kinase 4/6 axis.