Glyoxal exposure is associated with a spectrum of adverse health outcomes, including arterial injury. While existing evidence confirms that glyoxal targets oxidative stress and the mitogen-activated protein kinase pathway, the precise underlying mechanism of glyoxal-induced arterial damage remains elusive. In this study, we aimed to elucidate the molecular mechanism driving glyoxal-induced arterial injury through a combination of in vitro and in vivo experiments. In vitro, we treated human aortic endothelial cells with glyoxal, then performed cell viability assays, tandem mass tag (TMT)-based quantitative proteomics, parallel reaction monitoring (PRM), glutathione quantification for oxidative stress assessment, western blotting, and quantitative polymerase chain reaction (qPCR) analysis. In vivo, we administered glyoxal to C57BL/6 mice, after which we conducted biochemical, histological, and immunohistochemical assays to evaluate arterial pathological changes. In vitro, the proteomic profiling results were further validated via PRM and western blotting, which confirmed that glyoxal exposure induces energy metabolism dysfunction, aberrant vascular endothelial growth factor receptor (VEGF-R) expression, and signaling pathway inactivation. Consistent with the proteomic results, we detected reduced protein levels of VEGF-R (36.4 Our previous study demonstrated that oxidative stress and MAPK pathways are important targets of glyoxal (GX); however, the specific mechanism is not completely understood at present. Therefore, this study investigates the AI mechanism induced by GX based on the proteomics. These novel mechanisms include INS-R, IGF-R, and VEGF-R damage and systematic changes in the Wnt and ErbB signaling pathways. These injuries correspond to abnormal energy metabolism and function of HAECs, causing aberrant vascular function, inflammation, and abnormal vascular structure.
CircARHGAP10 is significantly downregulated in colorectal cancer (CRC), but its functional role and underlying molecular mechanism in CRC progression remain unelucidated. Here, we first detected circARHGAP10 expression in CRC tissues and adjacent normal tissues, as well as in CRC cell lines and normal intestinal epithelial cells. Functional analyses using CCK-8, EdU, colony formation, wound healing, and transwell assays demonstrated that circARHGAP10 overexpression notably inhibits CRC cell proliferation, migration, and invasion, while circARHGAP10 knockdown promotes these malignant phenotypes. Mechanistically, bioinformatic predictions, dual-luciferase reporter assays, and RNA immunoprecipitation (RIP) assays confirmed that circARHGAP10 acts as a competing endogenous RNA (ceRNA) to sponge miR-29a-5p, thereby upregulating the expression of its downstream target gene LPP. Furthermore, we found that circARHGAP10 modulates LPP, the Wnt/β-catenin signaling pathway and reverses epithelial-mesenchymal transition (EMT) through the miR-29a-5p, as validated by rescue experiments. Collectively, our findings reveal that circARHGAP10 functions as a tumor suppressor in CRC via two independent regulatory branches downstream of the circARHGAP10/miR-29a-5p cascade. On one hand, it restores LPP expression to block the malignant proliferation, migration and invasion of CRC cells; on the other hand, this signaling cascade mitigates excessive Wnt/β-catenin pathway activation and reverses EMT. This newly identified circRNA-centered regulatory network provides a promising novel therapeutic target for CRC intervention.
Curcumin, a natural polyphenol derived from turmeric, possesses potent antioxidant properties and represents a potential cardioprotective agent capable of mitigating myocardial oxidative stress associated with rheumatoid arthritis (RA). The aim of the study was to examine the cardioprotective effect of curcumin in an experimental model of rheumatoid arthritis. The study included 104 female Wistar albino rats, divided into 8 groups (n = 13 in each group): (1) CTRL (negative control, healthy rats without treatment), (2) CUR (positive control 1, curcumin 200 mg/kg three times a week for 4 weeks orally), (3) MTX (positive control 2, methotrexate 0.75 mg/kg twice a week for 4 weeks intraperitoneally), (4) RA (positive control 3, rats with Complete Freund’s adjuvant (CFA)-induced RA), (5) RA+pCUR (rats with preventive administration of curcumin + CFA-induced RA), (6) RA+tCUR (rats with CFA-induced RA + therapeutic administration of curcumin), (7) RA + MTX+pCUR (rats with preventive administration of curcumin + CFA-induced RA + methotrexate), (8) RA + MTX+tCUR (rats with CFA-induced RA + therapeutic administration of curcumin + methotrexate). Pro-oxidant levels were determined in coronary venous effluent, antioxidant enzyme activities were assessed in heart tissue homogenates, and histopathological analyses of the synovium and myocardium were performed. Curcumin, particularly in the therapeutic regimen and in combination with methotrexate, significantly reduced pro-oxidant levels and lipid peroxidation. Curcumin also increased superoxide dismutase activity and restored reduced glutathione levels, whereas improvement in catalase activity was observed predominantly in the therapeutic regimens. Histopathological analyses demonstrated that curcumin exerted a protective effect on both synovial and myocardial tissues, attenuating RA-induced structural damage, reducing edema, and preserving myocardial architecture. These findings suggest that curcumin improves myocardial redox homeostasis and may contribute to cardioprotection in experimental RA.
Celastrol (CEL), a quinonic methyl triterpenoid extracted from the rootstock of the plant Tripterygium wilfordii, which shows therapeutic potential in improving inflammatory diseases, tumor, metabolic diseases and neurodegenerative diseases. However, the role of CEL in acute inflammatory pain is still unclear. In our experiment, the inflammatory pain model induced by formalin was used to explore the mechanism and effect of CEL on pain regulation. The results showed that intraperitoneal injection (i.p.) or intragastric gavage (i.g.) of CEL (0.5 mg/kg and 1.0 mg/kg) obviously reduced the time of licking/biting the right hind paw in the formalin test, without affecting normal motor function or voluntary activity. Moreover, both naloxone (NLX, 1.0 mg/kg, co-injected with CEL) and nor-binaltorphimine (nor-BNI, 10 mg/kg, administrated 24 h prior to CEL) significantly antagonized the antinociceptive effect of CEL, indicating that the κ-opioid receptor was involved in the antinociceptive effect of CEL. In addition, CEL significantly up-regulated the gene expression and protein levels of dynorphin mRNA and Oprk1, while did not influence the expression of Penk, Pomc, Oprm1 and Oprd1. The protein expression levels of p-CaMKII, p-ERK and p-CREB were also significantly decreased in the CEL treated group. Moreover, the immunohistochemistry and immunofluorescence results demonstrated the numbers of positive c-Fos and GFAP in the dorsal horn (L4-6) were obviously reduced. However, all of these protein’s expression change induced by CEL were blocked by nor-BNI. Thus, our results indicate that CEL may produce analgesic effect through activating KOR, which subsequently reduce the expression of p-CaMKII/ERK/CREB and inhibit the activation of neural cells in the dorsal horn (L4-6).
Atopic dermatitis (AD) is primarily characterized by dysregulation of the immune system and abnormal inflammation. Dictamni Cortex is an herb used in traditional Chinese medicine to treat damp-heat skin diseases. To excavate the effective component of Dictamni Cortex for treating AD and explore the specific molecular mechanism. First, wogonin was screened from the active constituents of Dictamnus cortex, and CCL2 was identified as a candidate hub gene by constructing an herb-ingredient-gene network. The AD cell model was constructed using human epidermal keratinocytes (HEKs), and the AD mouse model was employed to investigate the therapeutic efficacy of wogonin. The effect of HEKs on neutrophil extracellular trap (NET) formation under pathological conditions was explored through co-culture experiments. The conditional medium containing NETs was used to treat HEKs to investigate AD-associated inflammatory alterations. Through bioinformatics means, miR-4667-5p was predicted as an upstream regulator targeting CCL2 mRNA, and this was validated via RNA pull-down and dual-luciferase reporter assays. CCL2 was significantly upregulated in the AD cell model, which mediated the functional damage of HEKs under AD pathological conditions. Wogonin not only rescued the cell viability of HEKs in a concentration-dependent manner by inhibiting CCL2 but also alleviated the AD-related inflammation in vitro and in vivo. Within the co-culture system, AD model cells recruited neutrophils and induced NET formation, which was suppressed following CCL2 knockdown. Overexpression of miR-4667-5p suppressed CCL2 expression by targeting its mRNA and alleviated inflammation in HEKs induced by NET-conditioned medium. This inflammation was also suppressed by wogonin treatment. The miR-4667-5p antagonist weakened this therapeutic effect of wogonin, which was rescued by the silencing of CCL2. Under AD pathological conditions, HEKs promoted NET formation, which in turn exacerbated inflammation within HEKs. Wogonin treatment rescued this inflammatory damage by modulating the miR-4667-5p/CCL2 pathway.
Myocardial fibrosis is characterized by excessive cardiac fibroblasts (CF) proliferation and extracellular matrix deposition. The p53 tumor suppressor is known to inhibit CF proliferation, while the polycomb protein Bmi1 negatively regulates p53. Ellagic acid (EA), a natural polyphenol, has demonstrated anti-fibrotic potential, but its mechanism of action in the heart remains unclear. This study investigated whether EA reduces myocardial fibrosis by inhibiting Bmi1 expression. A rat model of myocardial fibrosis was established using isoproterenol (ISO), and rat cardiac fibroblasts were treated with TGF-β to induce a pro-fibrotic phenotype in vitro. Echocardiography and histology confirmed that EA treatment improved cardiac function and reduced fibrosis in ISO-induced rats. In vitro, EA significantly inhibited TGF-β-induced CF proliferation, migration, and differentiation into myofibroblasts. Mechanistically, EA upregulated p53 and its downstream target p21, leading to G1/S cell cycle arrest. Molecular docking and dynamics simulations predicted that EA binds to Bmi1, and subsequent experiments showed EA treatment reduced Bmi1 protein levels. Using a Bmi1 inhibitor (PTC-209) and overexpression plasmids, we demonstrated that the anti-fibrotic effects of EA are mediated in part through inhibition of Bmi1, which in turn activates the p53 pathway. Ellagic acid reduces myocardial fibrosis, at least in part, through suppression of Bmi1 expression. This suppression is associated with relief of the inhibitory effect of Bmi1 on p53, leading to p53/p21 pathway activation, cell cycle arrest, and subsequent inhibition of cardiac fibroblasts proliferation, migration, and differentiation. These findings suggest that Bmi1 may represent a novel therapeutic target for EA treatment of myocardial fibrosis.
The salivary glands of hematophagous arthropods possess a remarkable molecular diversity, reflecting evolutionary adaptations to blood feeding lifestyle and vector-host interactions. In this study, we characterized the salivary-gland transcriptome of adult female Simulium pertinax, a medically important black fly species widely distributed in South America. Through RNA sequencing and bioinformatics analyses, we identified 15,708 high-quality transcripts, of which 1947 clusters were related to secreted proteins, distributed across 23 distinct functional families, including digestive enzymes, protease inhibitors, antimicrobial peptides, and hemostasis modulators. Phylogenetic analyses of selected hematophagy-associated families, including Kunitz-domain inhibitors, Apyrases, Kazal-type inhibitors, Serpins, and SVEP homologs, revealed a combination of conserved evolutionary relationships and lineage-specific diversification among simuliids. Notably, the SVEP (Simulium Vasodilator Erythema Protein) family was highly represented and appears to be restricted to black flies. Given that its biological role remains largely uncharacterized, we selected a novel SVEP variant for heterologous expression and functional characterization, which was designated as recombinant Simulium pertinax Vasoactive Protein (rSpVaP). This recombinant protein elicited vasodilator activity in the arterioles of the mouse cremaster muscle, as well as significant paw edema. These findings support rSpVaP as a biologically active SVEP-related salivary vasoactive protein in simuliids, underscoring its contribution to facilitating blood-feeding. Taken together, these results highlight the value of combining transcriptomic profiling with functional characterization to uncover the molecular adaptations underlying hematophagy in black flies, enhancing our understanding of vector-host interactions and the evolutionary pressures shaping the sialotranscriptome.
Mannose has emerged as a promising metabolic modulator capable of impairing tumor growth in phosphomannose isomerase (PMI)-deficient cancers. However, the molecular adaptations underlying the response of melanoma cells to mannose remain poorly understood. Here, we compared the phenotypic, proteomic, and phosphoproteomic responses of normal melanocytes (Melan-A) and metastatic melanoma cells (B16F10) cultured in glucose- or mannose-containing media. Both cell types exhibited similar phenotypic responses to mannose, including reduced growth, altered lactate production, and increased accumulation of acidic vesicles. Despite these shared phenotypes, proteomic and phosphoproteomic analyses revealed markedly distinct molecular adaptations. Mannose induced modest proteomic changes in normal melanocytes but extensive remodeling in metastatic melanoma cells, accompanied by profound phosphoregulatory remodeling. Substrate enrichment analysis based on curated signaling interactions identified phosphosubstrate signatures associated with predominantly homeostatic regulatory programs in normal melanocytes, whereas metastatic cells exhibited phosphosubstrate signatures associated with stress-responsive, survival, cell-cycle, and ubiquitin-mediated regulatory programs. Collectively, our findings demonstrate that similar phenotypic responses to mannose arise from fundamentally distinct molecular programs, highlighting cellular context as a key determinant of the adaptive response to mannose-induced metabolic stress in melanoma.
Cryptorchidism, characterized by the failure of one or both testes to descend into the scrotum, is the most prevalent urogenital malformation in male infants and an important risk element of male infertility and testicular cancer. The purpose of this study was to investigate the molecular mechanisms underlying cryptorchidism-induced testicular damage, with a focus on the role of USP35. Here we established a unilateral experimental cryptorchidism model of Sprague-Dawley rats by resetting the left testis to the abdominal cavity and suturing the inguinal canal, thus preventing its natural descent. The findings indicated that apoptosis and oxidative stress (OS) levels were increased in the undescended testis (UDT) compared to the descended testis (DT) group. Subsequent mRNA sequencing of DT and UDT tissues revealed that ubiquitin specific peptidase 35 (USP35) expression was downregulated in the UDT, implying that USP35 may be an important factor in the pathogenesis of cryptorchidism. To further investigate, an oxidative damage model was constructed using H2O2-treated GC-1 spermatogonia. USP35 expression was reduced in H2O2-exposed cells, while overexpression of USP35 attenuated H2O2-induced oxidative damage by inhibiting apoptosis and OS. Mechanistically, USP35 interacted with nuclear factor erythroid 2-related factor 2 (NRF2) and stabilized its protein expression via deubiquitination. Impairment of NRF2 function abolished the protective effect of USP35 against oxidative damage in H2O2-treated GC-1 spermatogonia. Overall, these results highlight the significance of the USP35-NRF2 axis in protecting spermatogonia from oxidative injury, suggesting that USP35 may be a potential therapeutic target against cryptorchidism-induced early germ cell damage.
Melanoma, particularly driven by activating mutations in the BRAF gene, has witnessed significant therapeutic advances through targeted therapies such as BRAF and MEK inhibitors. However, the development of acquired resistance remains a major clinical challenge, limiting long-term treatment efficacy. The tumor microenvironment (TME), especially tumor-associated macrophages (TAMs), plays a pivotal role in mediating immune evasion, angiogenesis, and drug resistance. Despite extensive research into TAM polarization and function, the precise molecular mechanisms underlying their contribution to therapy resistance remain incompletely understood. Here, we investigated the regulatory role of IGF1-IGF1R signaling in THP-1-derived macrophages and its impact on the sensitivity of co-cultured melanoma cells to the BRAF inhibitor PLX4032. We observed markedly upregulated IGF1 expression in both melanoma cells and THP-1-derived macrophages. Either IGF1R knockdown or pharmacological blockade of IGF1–IGF1R signaling in macrophages restrained the polarization of THP-1-derived macrophages. Both in vitro cellular assays and in vivo xenograft experiments verified that A375 cells pre-co-cultured with IGF1R-deficient macrophages were more sensitive to PLX4032 relative to those pre-co-cultured with IGF1R-proficient macrophages. Transcriptomic analysis reveals that IGF1-IGF1R signaling in THP-1-derived TAMs significantly regulates extracellular matrix (ECM) organization. Collectively, our results offer a new insight into IGF1-IGF1R signaling in THP-1-derived macrophages regulating ECM organization in TME and its potential role in indirectly modifying the tumor cells responsiveness to PLX4032.
Homeobox C9 (HOXC9) is aberrantly expressed in multiple malignancies; however, its functional role in esophageal squamous cell carcinoma (ESCC) remains elusive. This study investigated the expression, function, and underlying molecular mechanisms of HOXC9 in ESCC. HOXC9 was evaluated via immunohistochemistry in 118 ESCC and paired normal tissues. Stable cell lines with HOXC9 knockout, knockdown, and overexpression were established in KYSE70 and KYSE150 cells. Cell proliferation, apoptosis, mitochondrial membrane potential, and in vivo xenograft growth were assessed. Mechanistic studies were performed using RNA-seq, Western blotting, and PI3K inhibitor LY294002 (30 µM). IHC analysis revealed significantly elevated HOXC9 expression in ESCC versus paired adjacent normal tissues (high expression rate: 57.6
To explore the expression changes of fatty acid translocase (CD36) and carnitine palmitoyltransferase 1a (CPT1A) in rat cardiomyocytes after high-cervical spinal cord injury (SCI), and to clarify their relationship with Fatty acid metabolism disturbance and acute myocardial damage. Thirty-six male Sprague–Dawley rats were randomly assigned to control, sham-operation, and high-cervical SCI groups. A C7 contusion model was established by a modified Allen weight-drop method, with SCI rats sampled at 4, 12, 24, and 48 h post-injury. Myocardial ultrastructure was examined by transmission electron microscopy. The GSE45006 spinal cord transcriptomic dataset was reanalyzed for Cd36 and Cpt1a expression at 1 and 3 days after SCI. Cd36 mRNA and CPT1A protein levels in cardiac tissue were quantified by qRT-PCR and Western blotting, respectively. TEM showed progressively worsening myocardial ultrastructural injury in SCI rats from 4 to 48 h, manifesting as mitochondrial swelling, myofilament disarray, and intracellular edema. Bioinformatics analysis of injured spinal cord tissue showed that Cd36 was significantly upregulated at both 1 and 3 days after SCI, whereas Cpt1a was upregulated at 1 day and returned toward the sham level by 3 days. In cardiac tissue, Cd36 mRNA was significantly elevated at 4 h and reduced at 48 h, and CPT1A protein showed a parallel biphasic trend over the same period in the SCI group. No significant differences were detected between the control and sham groups. Dynamic dysregulation of CD36/CPT1A is associated with myocardial fatty acid metabolic disturbance after high-cervical SCI and may be involved in acute myocardial injury. CD36 and CPT1A may serve as potential targets for the prevention and treatment of SCI-induced myocardial dysfunction. High-cervical SCI is associated with biphasic dysregulation of CD36/CPT1A, paralleling myocardial fatty acid metabolic disturbance and acute injury. CD36 and CPT1A are potential therapeutic targets for SCI-induced myocardial dysfunction. Created with BioGDP.com
Osteoporosis is a bone disorder characterized by low bone mass density and impaired microarchitecture that is accompanied by multiple metabolic disorders, especially metabolic dysfunction-associated steatotic liver disease (MASLD). Inulin has been proposed as a treatment for MASLD, but its role in MASLD-related bone loss has not been determined. This study aimed to investigate the effect of inulin on MASLD-induced bone loss and the underlying mechanisms. To establish a MASLD model, C57BL/6 mice were fed a high-fat high-fructose high-glucose diet for 16 weeks. Inulin treatment was administered by incorporating 5
Rheumatoid arthritis (RA) is characterized by persistent synovial inflammation, oxidative imbalance, and progressive joint destruction. Celastrol (CEL), a bioactive triterpenoid derived from Tripterygium wilfordii, exhibits anti-inflammatory and antioxidant activities, but its mechanism of action in RA remains incompletely defined. This study investigated whether the Nrf2/HO-1/GPX4 pathway is involved in the protective effects of CEL in RA. Collagen-induced arthritis (CIA) rats were treated with CEL or methotrexate for 4 weeks. Arthritis severity, serum inflammatory cytokines, histopathological injury, oxidative stress, and pathway-related proteins were evaluated. In vitro, TNF-α-stimulated fibroblast-like synoviocytes (FLS) were used to assess proliferation, migration, invasion, apoptosis, inflammatory responses, and ROS/Fe²⁺ accumulation. tBHQ and Brusatol were used as pharmacological pathway-modulating probes in functional and rescue experiments. CEL significantly reduced paw swelling, arthritis scores, and serum IL-1β, IL-6, and TNF-α levels in CIA rats, and attenuated synovial hyperplasia, cartilage damage, collagen loss, and osteoclast activation. CEL also increased Nrf2, HO-1, and GPX4 expression while reducing ROS and Fe2+ accumulation in ankle-joint-derived single-cell suspensions. In TNF-α-stimulated rat FLS, CEL suppressed proliferation, migration, and invasion, increased apoptosis under inflammatory stimulation, and alleviated inflammatory and oxidative stress responses. These protective effects were pharmacologically enhanced by tBHQ and attenuated by Brusatol co-treatment. These findings suggest that CEL alleviates synovial inflammation and joint injury in experimental RA in association with Nrf2/HO-1/GPX4 pathway activation and suppression of TNF-α-stimulated rat FLS activation, supporting further investigation of CEL as a candidate antioxidant-oriented therapeutic agent for RA.
Osteoporosis is a prevalent chronic bone disease characterized by excessive osteoclast-mediated bone resorption. Liensinine (LIE), a plant-derived alkaloid, has shown anti-inflammatory potential but its role in bone resorption remains rarely investigated. This study aimed to predict the anti-osteoclastogenic mechanism of LIE via network pharmacology and evaluate the therapeutic potential of endothelial cell-derived extracellular vesicles loaded with LIE (LIE@EVs). In this study, extracellular vesicles derived from endothelial cells were isolated, identified, and utilized to encapsulate LIE. The physicochemical properties and internalization of LIE@EVs were assessed. Core pathways of LIE were predicted using network pharmacology and molecular docking. Furthermore, both in vitro and in vivo experiments were conducted to evaluate its inhibitory effects on RANKL-induced osteoclastogenesis and bone resorption. LIE@EVs exhibited characteristics of extracellular vesicles and enhanced delivery of LIE to osteoclasts. Functional assays demonstrated that LIE@EVs significantly inhibited osteoclast differentiation and bone resorption compared to free LIE. Network pharmacology identified the NF-κB and calcium signaling pathways as the potential targets of LIE. Mechanistically, LIE@EVs effectively suppressed the activation of the NF-κB signaling pathway and calcium oscillations, ultimately downregulating osteoclast-specific gene expression. These findings exhibit the potential therapeutic significance of LIE@EVs in bone loss diseases and warrant further exploration in clinical applications for osteoporosis treatment.
This study developed an engineered Lactobacillus plantarum-derived vesicle system for oral delivery of interleukin-22 mRNA (IL-22 mRNA) and evaluated its effects on epithelial barrier repair in DSS-induced colitis. The vesicle formulation, termed P-OMVs@IL-22 m, was prepared from L. plantarum protoplasts through lysozyme treatment, ultrasonic disruption, OptiPrep density-gradient purification, and incubation-based mRNA loading. The formulation achieved an IL-22 mRNA encapsulation efficiency of 72.43 ± 6.60 Mouse-based schematic of oral P-OMVs@IL-22 m administration in DSS-induced colitis, gastrointestinal retention, epithelial uptake, IL-22 expression, STAT3 activation, restoration of tight junction proteins and MUC2, and attenuation of intestinal inflammation.
Lysophosphatidic acid (LPA) receptor signaling contributes to the regulation of cancer cell functions. Lactic acid serves not only as an important energy source for cancer cells but also activates various signaling pathways that influence cancer cell behavior. To investigate the roles of LPA receptors in the malignant behavior of pancreatic cancer PANC-1 cells, PANC-LA1 and PANC-LA5 cells were generated by culturing PANC-1 cells with 1 or 5 mM lactic acid, respectively, for approximately 3 months. LPAR1, LPAR3, and LPAR5 expression levels expression levels were elevated, whereas LPAR2 expression was reduced in PANC-LA1 and PANC-LA5 cells compared with PANC-1 cells. PANC-1 cell growth was inhibited by LPA. Conversely, PANC-LA1 cell growth remained unchanged in response to LPA, and LPA increased PANC-LA5 cell growth. The motility of PANC-LA1 and PAN-LA5 cells was markedly elevated compared with that of PANC-1 cells. LPA further enhanced PANC-LA1 and PAN-LA5 cell motility but did not affect PANC-1 cell motility. In the presence of LPA, AM966 inhibited, whereas TC LPA5 4 stimulated PANC-LA5 cell growth and PANC-LA1 cell motility. Although GRI-977,143 increased PANC-LA5 cell growth, it reduced PANC-LA1 cell motility. (2S)-OMPT increased PANC-LA5 cell growth and PANC-LA1 cell motility. When cells were cultured in serum-free media, the viability of PANC-LA1 and PANC-LA5 cells was higher compared with that of PANC-1 cells. PANC-LA5 cell growth and PANC-LA1 cell motility were inhibited by AZD3965 and galloflavin. These findings indicate that LPA receptor signaling is involved in the regulation of PANC-1 cell behavior induced by long-term lactic acid exposure, with LPA1 and LPA3 acting as major positive regulators of proliferation and motility following long-term lactic acid exposure, whereas LPA2 and LPA5 exert distinct regulatory effects.
Although Angiotensin-(1-7) [Ang-(1-7)], a non-classical peptide of the renin-angiotensin system (RAS), is widely recognized for alleviating cardiovascular stress through activation of the Mas receptor. we identified a largely Mas receptor-independent pathway that expands its therapeutic potential in cardiac diseases. Our study demonstrated that Ang-(1-7) stimulated cardiomyocytes to release small extracellular vesicles (sEVAng-(1-7)), which exhibit enhanced cardioprotective effects compared with Ang-(1-7) administration alone. In both wild-type C57BL/6J mice and cardiac-specific Mas receptor knockout (cMas-KO) mice subjected to myocardial ischemia/reperfusion injury (MIRI), we found that enriched sEVAng-(1-7) preserved cardiac function by attenuating cardiomyocyte apoptosis and oxidative stress following MIRI. These protective effects remained evident even when Mas receptor signaling was pharmacologically inhibited or genetically ablated. Moreover, in wild-type mice with MIRI, enriched sEVAng-(1-7) provided significantly greater therapeutic efficacy in improving cardiac function than direct administration of Ang-(1-7). In AC16 human cardiomyocytes subjected to hypoxia/reoxygenation (H/R), enriched sEVAng-(1-7) enhanced Akt2 phosphorylation, increased sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) activity, maintained calcium homeostasis, suppressed endoplasmic reticulum stress signaling (CHOP and p-JNK), and protected AC16 cells against H/R-induced apoptosis. sEVAng-(1-7) also attenuated oxidative stress under these conditions. Importantly, the protective effects of sEVAng-(1-7) remained after Mas receptor knockdown. Separately, Akt2 knockdown (Akt2KD) substantially weakened these benefits. These findings identify sEVAng-(1-7) as an promising therapeutic candidate, offering an effective, largely Mas-independent mechanism to combat MIRI with cardiomyocyte protection.
Peritoneal dialysis (PD) is a renal replacement therapy for patients with end-stage renal disease, but long-term treatment often leads to peritoneal fibrosis, compromising dialysis efficacy. This study aimed to elucidate the role of ferroptosis in peritoneal fibrosis (PF) and to clarify the regulatory function of the voltage-dependent anion channel 2 (VDAC2). Mesothelial cell subpopulations were identified from the single-cell dataset GSE130888, followed by differential gene expression analysis to screen ferroptosis-related genes. In vitro and in vivo models were used to validate the involvement of ferroptosis in PF. siRNA-mediated VDAC2 knockdown, combined with the ROS scavenger NAC, was applied to assess its role in regulating mitochondrial ROS during ferroptosis and fibrosis. Single-cell analysis revealed ferroptosis-related gene alterations in both short-term (ST) and long-term (LT) PD, with VDAC2 expression positively associated with dialysis duration. Erastin significantly reduced MET‑5 A cell viability, increased apoptosis, elevated ROS and lipid peroxidation, decreased GSH levels, and upregulated VDAC2, ACSL4, and fibrosis‑related proteins. These effects were reversed by Fer‑1. In vivo, Erastin exacerbated peritoneal dysfunction, collagen deposition, and tissue iron accumulation in mice, and caused complete collapse of cellular architecture, dissolution of organelles, nuclear envelope rupture, and chromatin fragmentation, whereas Fer‑1 improved peritoneal pathology and iron deposition. VDAC2 knockdown restored MET‑5 A cell viability, reduced ROS and lipid peroxidation, increased GSH levels, and downregulated ferroptosis- and fibrosis-related proteins; co‑treatment with NAC further enhanced these protective effects. VDAC2 promotes peritoneal mesothelial cell injury and fibrosis by driving mitochondrial ROS-dependent ferroptosis. Targeting VDAC2 or inhibiting ferroptosis may provide a potential therapeutic strategy to delay peritoneal dialysis-associated fibrosis.
Early life overnutrition can promote lasting metabolic and cardiovascular dysfunction. This study evaluated the effects of post-weaning exercise training on cardiac mitochondrial function, oxidative status, and mitochondrial-related gene expression in rats. After overnutrition period, male Wistar rats were assigned to sedentary or trained groups. The exercise protocol consisted of moderate-intensity treadmill running for 4 weeks. Exercise capacity was assessed before and after the intervention through a progressive running test to determine maximal running velocity (Vmax). Body weight was monitored and after euthanasia, fresh mitochondrial fractions were isolated from the left ventricle by differential centrifugation. Mitochondrial respiration was measured using a Clark-type oxygen electrode. Citrate synthase activity, swelling, reactive oxygen species (ROS) production, lipid peroxidation (MDA), protein carbonyls, redox status (NAD/NADH and GSH/GSSG), total thiol content, and the expression of PGC-1α, TFAM, FIS1, OPA1, and UCP2 were evaluated. Data were analyzed using Student’s t-test and two-way ANOVA followed by Tukey’s post hoc test. Exercise training increased Vmax (p = 0.014), indicating improved exercise capacity. In left ventricular mitochondria, training enhanced respiratory efficiency and citrate synthase activity (p = 0.028), reduced ROS production (p = 0.035), and attenuated oxidative damage, as shown by lower MDA (p = 0.011) and protein carbonyl levels (p = 0.022). However, mitochondrial swelling analyses did not differ between groups. Antioxidant defenses were strengthened, with an increased GSH/GSSG ratio (p = 0.009) and preserved thiol content (p = 0.041). Exercise also upregulated PGC-1α, TFAM, and FIS1 expression. Post-weaning exercise improves cardiac mitochondrial function, reduces oxidative stress, and modulates mitochondrial dynamics in rats exposed to early life overnutrition.
