RESUMO
JOURNAL/mgres/04.03/01612956-202609000-00005/figure1/v/2026-01-06T135433Z/r/image-tiff Sevoflurane is a new type of halogen inhalation anesthetic gas with a rapid induction and emergence. It is widely used for general anesthesia. Previous studies have demonstrated that sevoflurane postconditioning alleviates cerebral ischemia-reperfusion injury and enhances the tolerance of the brain to ischemia and hypoxia. However, whether sevoflurane postconditioning can reduce cerebral ischemia-reperfusion injury caused by hemorrhagic shock and resuscitation and the underlying mechanism are unclear. The present study established cerebral ischemia-reperfusion injury models through an in vivo hemorrhagic shock and resuscitation method in C57BL/6 mice and an in vitro oxygen-glucose deprivation/reoxygenation method in HT22 cells. After hemorrhagic shock and resuscitation treatment, the mice developed significant spatial learning and memory impairments accompanied by aggravated cerebral infarction, whereas sevoflurane postconditioning significantly improved these effects. After in vitro oxygen-glucose deprivation/reoxygenation, the survival rate of HT22 cells was decreased, the apoptosis rate was increased, the expression of silent information regulatory factor 1 was decreased, and the expressions of hypoxia-inducible factor 1α, NOD-like receptor protein 3, gasdermin D, caspase-1, and interleukin-1ß were increased. Sevoflurane postconditioning inhibited oxygen-glucose deprivation/reoxygenation-induced changes. Following silent information regulatory factor 1 knockdown by small interfering RNA, the cytoprotective effects of sevoflurane postconditioning were significantly attenuated. These findings suggest that the anesthetic gas sevoflurane postconditioning ameliorates hemorrhagic shock and resuscitation-induced cognitive impairment. This may be mediated by the silent information regulatory factor 1/hypoxia-inducible factor 1α/NOD-like receptor protein 3 pathway.
Assuntos
Anestésicos Inalatórios , Disfunção Cognitiva , Pós-Condicionamento Isquêmico , Ressuscitação , Sevoflurano , Choque Hemorrágico , Sevoflurano/farmacologia , Sevoflurano/uso terapêutico , Animais , Choque Hemorrágico/complicações , Camundongos Endogâmicos C57BL , Camundongos , Ressuscitação/efeitos adversos , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/tratamento farmacológico , Anestésicos Inalatórios/farmacologia , Anestésicos Inalatórios/uso terapêutico , Masculino , Apoptose/efeitos dos fármacos , Traumatismo por Reperfusão , Linhagem Celular , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismoRESUMO
Acute kidney injury (AKI) is a global health concern with various etiologies, including ischemia-reperfusion injury, sepsis, and nephrotoxic agents such as cisplatin. Cisplatin-induced nephrotoxicity is associated with lysosomal damage, but the underlying mechanisms remain unclear. This study aimed to elucidate the role of lysosomal damage in cisplatin-induced cell death in tubular epithelial cells (TECs). AKI models were induced using cisplatin both in vivo and in vitro. Mouse kidney morphology and function were assessed using biochemical assays, immunohistochemistry, and histological staining techniques such as HE, PAS, and TUNEL. RNA sequencing analysis and pharmacological interventions were used to investigate the specific mechanism in cisplatin-induced TECs injury. The distribution and expression of lysosomes and calcium (Ca2+) were measured through immunofluorescence staining, Western blotting, and confocal microscopy. RNA sequencing analysis revealed a notable role of Ca2+ signaling pathways in cisplatin-induced nephrotoxicity. We demonstrated that cisplatin exposure induced significant lysosomal abnormalities, including altered distribution, morphology, and increased Ca2+ leakage. This dysregulation of lysosomal Ca2+ homeostasis was closely correlated with TECs apoptosis. Mechanistically, we show that lysosome Ca2+ release activates calcineurin, thereby triggering apoptosis in TECs. Preliminary data indicate that inhibiting lysosomal Ca2+ release through targeting TRPML1 may mitigate cisplatin-induced AKI. Our study reveals a lysosomal Ca2+-calcineurin pathway that contributes to cisplatin-induced nephrotoxicity and offers potential therapeutic targets.
Assuntos
Injúria Renal Aguda , Cálcio , Cisplatino , Células Epiteliais , Túbulos Renais , Lisossomos , Canais de Potencial de Receptor Transitório , Animais , Cisplatino/toxicidade , Cisplatino/efeitos adversos , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Camundongos , Lisossomos/metabolismo , Lisossomos/efeitos dos fármacos , Cálcio/metabolismo , Túbulos Renais/metabolismo , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/patologia , Homeostase/efeitos dos fármacos , Canais de Potencial de Receptor Transitório/metabolismo , Canais de Potencial de Receptor Transitório/genética , Masculino , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Camundongos Endogâmicos C57BL , Apoptose/efeitos dos fármacos , Antineoplásicos/toxicidadeRESUMO
Efferocytosis is a process in which specialized phagocytes, mainly macrophages, are responsible for recognizing and removing apoptotic cells from the inflammatory site, leading to tissue resolution, and preventing chronic inflammation. Herein we describe in detail a robust efferocytosis assay based on morphological criteria and flow cytometric for the quantification of efferocytic events and characterization of the engulfing efficiency of apoptotic cells by macrophage in vitro and in vivo, which can be used in many applications, including the development of genetic and pharmacological therapeutic targeting inflammation resolution.
Assuntos
Apoptose , Macrófagos , Fagocitose , Animais , Macrófagos/metabolismo , Macrófagos/citologia , Citometria de Fluxo/métodos , Camundongos , Humanos , Inflamação/patologiaRESUMO
Autophagy is an evolutionarily conserved cellular mechanism in eukaryotes that plays an important role in the maintenance of cellular homeostasis. The autophagy process maintains protein homeostasis by recycling damaged organelles and degrading many long-lived or damaged proteins through lysosomes in coordination with the ubiquitin-proteasome system. Cytokines are low molecular weight secreted proteins that regulate a broad range of biological activities. For instance, pro-inflammatory cytokines such as tumor necrosis factor-α (TNFα) induce inflammation, autophagy, and apoptotic cell death. In this chapter, we discuss experimental techniques such as immunoblotting and fluorescence microscopy that can be utilized to measure autophagy in response to TNFα treatment.
Assuntos
Autofagia , Neoplasias , Fator de Necrose Tumoral alfa , Autofagia/efeitos dos fármacos , Humanos , Fator de Necrose Tumoral alfa/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Neoplasias/metabolismo , Neoplasias/patologia , Microscopia de Fluorescência/métodos , Linhagem Celular Tumoral , Immunoblotting/métodos , Apoptose/efeitos dos fármacosRESUMO
Hepatic encephalopathy (HE), which develops as a result of liver failure, is an important neurological disorder involving inflammation and oxidative damage, with apoptosis and autophagy supported mainly by experimental evidence. In the current research, we researched the protective effects of oleuropein (OLE) in a thioacetamide (TAA)-induced HE model, particularly through the PI3K/Akt/mTOR signalling pathway. To execute the planned experimental design, Sprague Dawley rats (n = 28) were divided into four groups: Control, OLE, TAA, and TAA + OLE. OLE was administered orally (50 mg/kg) during a 14-day period, followed by intraperitoneal TAA (50 mg/kg) for 14 days in the TAA groups. Behavioral tests (open field and Y-maze) were used to determine cognitive and anxiety-like disorders in the rats. Oxidative stress indicators (MDA, SOD, and GSH), pro-inflammatory cytokines (IL-1ß, IFN-γ, and TNF-α), autophagic and apoptotic processes (Caspase-3, Bcl-2, Beclin-1, LC3), PI3K/Akt/mTOR pathway proteins, and AQP4 levels were analyzed in the serum and tissue. Histopathological evaluation was used to evaluate tissue damage in the liver and brain. The results indicated that the TAA-activated PI3K/Akt/mTOR pathway was suppressed by OLE, oxidative damage, autophagy, apoptosis, and inflammation were reduced, and behavioral and histological improvements were achieved. These results suggest that OLE offers hepatoprotective effects and ameliorates HE-associated brain injury via the PI3K/Akt/mTOR pathway.
Assuntos
Apoptose , Autofagia , Comportamento Animal , Encefalopatia Hepática , Glucosídeos Iridoides , Iridoides , Estresse Oxidativo , Animais , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Ratos Sprague-Dawley , Serina-Treonina Quinases TOR/metabolismo , Autofagia/efeitos dos fármacos , Autofagia/fisiologia , Ratos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Glucosídeos Iridoides/farmacologia , Glucosídeos Iridoides/uso terapêutico , Masculino , Fosfatidilinositol 3-Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Encefalopatia Hepática/induzido quimicamente , Encefalopatia Hepática/metabolismo , Encefalopatia Hepática/tratamento farmacológico , Tioacetamida/toxicidade , Inflamação/metabolismo , Inflamação/tratamento farmacológico , Comportamento Animal/efeitos dos fármacos , Iridoides/farmacologia , Iridoides/uso terapêuticoRESUMO
BACKGROUND: Sepsis-associated acute respiratory distress syndrome (ARDS) is a severe inflammatory lung disorder with high mortality. Bruceine A (BA), a quassinoid from Brucea javanica, exhibits anti-inflammatory and immunomodulatory activities, but its role in ARDS is unclear. OBJECTIVES: This study evaluated the protective effects of BA in lipopolysaccharide (LPS)-induced ARDS and explored its underlying mechanisms. METHODS: Thirty-six C57BL/6 mice were randomized into four groups: Control, LPS, LPS+BA and LPS+dexamethasone (Dex). Lung injury was assessed by histopathology, wet/dry weight ratio and TUNEL assay. Cytokine levels (TNF-α, IL-6, IL-1ß, IL-10) were measured by ELISA. Macrophage polarization markers (iNOS, COX-2, Arg-1, YM1, CD206) and NF-κB pathway proteins were evaluated using immunohistochemistry and Western blotting. RESULTS: BA significantly alleviated LPS-induced lung injury, reducing edema, tissue damage and alveolar apoptosis. It suppressed proinflammatory cytokines while enhancing IL-10. BA shifted macrophage polarization from proinflammatory M1 toward anti-inflammatory M2 phenotypes. Furthermore, BA inhibited NF-κB activation, evidenced by reduced phosphorylated p65 and restored IκBα levels. These effects were comparable to Dex. CONCLUSION: BA protects against LPS-induced ARDS in mice by modulating cytokine release, promoting M2 macrophage polarization and suppressing NF-κB activation. These findings suggest BA as a promising natural immunomodulatory agent for inflammatory lung diseases.
Assuntos
Anti-Inflamatórios , Lesão Pulmonar , Macrófagos , NF-kappa B , Quassinas , Síndrome do Desconforto Respiratório , Sepse , Animais , Camundongos Endogâmicos C57BL , Sepse/complicações , Sepse/tratamento farmacológico , Sepse/metabolismo , Síndrome do Desconforto Respiratório/tratamento farmacológico , Síndrome do Desconforto Respiratório/metabolismo , Síndrome do Desconforto Respiratório/patologia , Síndrome do Desconforto Respiratório/etiologia , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Lipopolissacarídeos , Quassinas/farmacologia , Quassinas/uso terapêutico , Masculino , Citocinas/metabolismo , Anti-Inflamatórios/farmacologia , Modelos Animais de Doenças , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pulmão/metabolismo , Apoptose/efeitos dos fármacos , Lesão Pulmonar/tratamento farmacológico , Lesão Pulmonar/metabolismoRESUMO
BACKGROUND: Paclitaxel is used in oral cancer treatment, but drug sensitivity remains a concern. CHFR has been implicated in tumor regulation, yet its role in modulating paclitaxel sensitivity in oral cancer requires further investigation. OBJECTIVES: This study aimed to evaluate the effect of CHFR on enhancing the drug sensitivity of paclitaxel in oral cancer cells. METHODS: A rat oral tumor model was established, followed by paclitaxel intervention. Observations included tongue tissue morphology, immune function, cell cycle, apoptosis, and the expression levels of NF-κB and CHFR proteins and mRNAs. RESULTS: The modeling success rate was 100%, with visible tongue masses and ulceration. CHFR protein expression increased in the CHFR mimic group. The high-dose paclitaxel group showed the highest immune indices, increased G0/G1 phase cell proportion, and significantly decreased tumor cell viability. The CHFR mimic group exhibited the smallest tumor volume, marked tumor cell death, and active proliferation. CHFR downregulated NF-κB expression; CHFR mRNA was higher, and NF-κB mRNA lower, compared to the high-dose paclitaxel and CHFR mimic groups. CONCLUSION: CHFR enhances paclitaxel sensitivity in oral cancer cells by downregulating NF-κB, effectively inhibiting tumor cell activity and suppressing tumor progression.
Assuntos
Antineoplásicos Fitogênicos , Resistencia a Medicamentos Antineoplásicos , Neoplasias Bucais , Paclitaxel , Paclitaxel/farmacologia , Animais , Neoplasias Bucais/tratamento farmacológico , Neoplasias Bucais/patologia , Neoplasias Bucais/metabolismo , Neoplasias Bucais/genética , NF-kappa B/metabolismo , NF-kappa B/genética , Linhagem Celular Tumoral , Apoptose/efeitos dos fármacos , Antineoplásicos Fitogênicos/farmacologia , Ratos , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Masculino , Humanos , Sobrevivência Celular/efeitos dos fármacos , Ratos Sprague-Dawley , Ciclo Celular/efeitos dos fármacosRESUMO
BACKGROUND: Luffa cylindrica flower has been used to treat haemorrhoids and breast hyperplasia, but the mechanism is unclear. OBJECTIVES: In this study, the antiproliferation activity of L. cylindrica flower extract (LCFE) was tested in breast cancer cells. METHODS: Following LCFE treatment, the CCK-8 assay, Hoechst staining and wound healing assay were used to evaluate cell proliferation, apoptotic cell morphology, and cell migration, respectively. qRT-PCR and western blot were used to analyze the expression of genes and proteins involved in the apoptosis and autophagy pathways. LC-MS was performed to characterize chemical constituents of LCFE. RESULTS: CCK-8 and wound healing assays revealed that LCFE suppressed the proliferation and migration of MCF-7 and MDA-MB-231 cells. In these two breast cancer cells, the extract treatment induced apoptotic morphological changes. LCFE treatment induced apoptosis by upregulating ZFP36 and BNIP3 expression, while downregulating Bcl-2 expression in MCF-7 cells. LCFE increases ZFP36 expression while decreasing BMP4 expression in MDA-MB-231 cells, promoting apoptosis. Meanwhile, LCFE treatment induced autophagy by increasing VMP1 expression and activating LC3 in MCF-7 cells. It also triggered autophagy by decreasing TBC1D14 expression, increasing ATG5 and VAMP8 expression, and activating LC3 in MDA-MB-231 cells. CONCLUSION: LCFE exerts an anti-tumor effect by activating apoptosis and autophagy processes in breast cancer cells, while having low cytotoxicity for normal breast cells, highlighting the potential of LCFE as a natural agent for cancer treatment.
Assuntos
Antineoplásicos Fitogênicos , Apoptose , Autofagia , Neoplasias da Mama , Flores , Luffa , Extratos Vegetais , Humanos , Autofagia/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/isolamento & purificação , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Neoplasias da Mama/metabolismo , Flores/química , Feminino , Células MCF-7 , Proliferação de Células/efeitos dos fármacos , Luffa/química , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Movimento Celular/efeitos dos fármacos , Linhagem Celular TumoralRESUMO
BACKGROUND: Spinal cord injury (SCI) is a severe condition causing sensory, motor, and autonomic dysfunctions, with over 759,000 patients and 66,374 new cases yearly in China. Secondary injury, driven by inflammation and apoptosis, hinders neurorestoration, making treatment challenging. Schisantherin B (SCHB), an active component of the traditional Chinese medicine Schisandra chinensis, has anti-inflammatory and anti-apoptotic effects in cerebrovascular and neurodegenerative diseases but its role in SCI remains unstudied. OBJECTIVES: This study aimed to investigate SCHB's therapeutic effects on SCI and clarify its underlying molecular mechanism, focusing on the PI3K/AKT signaling pathway. METHODS: In vitro, H2O2-induced PC12 cell apoptosis models were treated with different SCHB concentrations; cell viability (microplate reader), apoptosis (flow cytometry, immunofluorescence for cleaved caspase-3), and PI3K/AKT activation (immunofluorescence) were detected. In vivo, mouse SCI models (12.5g weight-drop contusion) received 15mg/kg SCHB intravenously; motor function (Basso Mouse Scale, footprint analysis), tissue damage (HE/Nissl staining), apoptosis (TUNEL staining), inflammation (ELISA for TNF-α/IL-1ß/IL-6/IL-10), and PI3K/AKT activation (Western blot, immunofluorescence) were assessed. RESULTS: SCHB (25µM in vitro) increased PC12 cell viability (66.15% vs. 40.86% in H2O2 group), reduced apoptosis (5.84% vs. 10.81%), and upregulated PI3K/AKT proteins. In mice, SCHB improved BMS scores (21/28 days post-injury), increased stride length/width, reduced spinal cord cavity size, preserved motor neurons, lowered pro-inflammatory cytokines (TNF-α/IL-1ß/IL-6), elevated IL-10, and activated the PI3K/AKT pathway. CONCLUSION: SCHB exerts therapeutic effects on SCI by inhibiting inflammation and apoptosis via activating the PI3K/AKT signaling pathway, supporting its potential as a candidate for SCI treatment.
Assuntos
Ciclo-Octanos , Lignanas , Fármacos Neuroprotetores , Fosfatidilinositol 3-Quinase , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Transdução de Sinais , Traumatismos da Medula Espinal , Animais , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/enzimologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células PC12 , Transdução de Sinais/efeitos dos fármacos , Ratos , Apoptose/efeitos dos fármacos , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Ciclo-Octanos/farmacologia , Lignanas/farmacologia , Modelos Animais de Doenças , Sobrevivência Celular/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Masculino , Medula Espinal/efeitos dos fármacos , Medula Espinal/patologia , Medula Espinal/enzimologia , Fosfatidilinositol 3-Quinase/metabolismo , Peróxido de HidrogênioRESUMO
Mitochondria are key players in regulating cellular metabolism. Short noncoding RNAs, or microRNAs (miRNAs), have become significant modulators of gene expression and cellular functions. Recent research has demonstrated the presence and activity of mitochondrial miRNAs (MitomiRs) in various cell types, including blood cells. The role of MitomiRNAs in metabolic reprogramming throughout blood cell formation, including their biosynthesis, function, and possible therapeutic implications, was discussed in this chapter. The discovery of mitochondrial microRNAs (MitomiRs) transformed our understanding of gene regulation in these critical organelles. These MitomiRs were discovered as they were influencing mitochondrial gene expression, with considerable effects on various cellular functions like oxidative stress responses, production of energy, and apoptosis during blood cell development.
Assuntos
Células Sanguíneas , MicroRNAs , Mitocôndrias , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Mitocôndrias/genética , Mitocôndrias/metabolismo , Estresse Oxidativo/genética , Animais , Células Sanguíneas/metabolismo , Células Sanguíneas/citologia , Regulação da Expressão Gênica , Apoptose/genética , Metabolismo Energético/genética , Reprogramação Celular , Reprogramação MetabólicaRESUMO
Protein tyrosine phosphatase nonreceptor 18 (PTPN18) is widely expressed in breast cancer (BC) cell lines. Additionally, high levels of PTPN18 facilitate an improved overall survival and prognosis in patients with BC. However, the effects and mechanisms of PTPN18 in BC remain unclear. In the present study, it was found that PTPN18 serves a tumor suppressor role in BC cells by promoting apoptosis, inhibiting proliferation and metastasis and inducing cell cycle arrest. Bioinformatics analysis showed that PTPN18 was significantly negatively correlated with the cell cycle and downregulated cyclin E expression, which was consistent with the experimental results. Subsequent coimmunoprecipitation assay results showed that PTPN18 could bind to cyclin E and promote its degradation through the ubiquitinproteasome pathway. Moreover, the addition of cyclin E2 did not reduce the binding of PTPN18 to cyclin E1. In the present study, the signaling pathways involved in cell cycle regulation were further investigated and it was found that PTPN18 may regulate the expression levels of cyclindependent kinase (CDK) inhibitor 1A and CDK inhibitor 1B proteins through phosphatidylinositol 3kinase/protein kinase B signaling pathway, which leads to cell cycle arrest and tumor inhibition in BC. Thus, analysis of the tumor suppressor mechanism of PTPN18 not only helps us to understand its biological function but also provides a theoretical basis for the development of new therapeutic strategies for BC.
Assuntos
Neoplasias da Mama , Ciclina E , Proteínas Tirosina Fosfatases não Receptoras , Humanos , Neoplasias da Mama/patologia , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Feminino , Ciclina E/metabolismo , Ciclina E/genética , Regulação Neoplásica da Expressão Gênica , Proliferação de Células , Linhagem Celular Tumoral , Proteínas Tirosina Fosfatases não Receptoras/metabolismo , Proteínas Tirosina Fosfatases não Receptoras/genética , Transdução de Sinais , Apoptose , Prognóstico , Regulação para Baixo , Genes Supressores de Tumor , Células MCF-7 , Pontos de Checagem do Ciclo Celular , Proteínas OncogênicasRESUMO
Kidney injury is a common complication of deep hypothermic circulatory arrest (DHCA) in patients undergoing cardiac surgery. However, the role of competitive endogenous RNA (ceRNA) networks in mediating DHCA-induced kidney injury has not been fully elucidated. In the present study, we aimed to systematically analyze ceRNA networks and identify a novel lncRNA, MSTRG.16386.1, that promotes DHCA-induced kidney injury by regulating the miR-466b-5p/Sprouty RTK signaling antagonist 2 (Spry2) axis. Kidney injury induced by DHCA was confirmed using a rat model of cardiopulmonary bypass (CPB) with or without DHCA. In addition, lncRNA and mRNA sequencing of kidney tissues revealed 309 specific shared differentially expressed mRNAs (DEGs) and 439 differentially expressed lncRNAs (DELs) in the kidneys of CPB+DHCA rats compared with those of SHAM and CPB rats. Differentially expressed miRNAs (DEMs) were predicted by coexpression and binding site analysis. A ceRNA network consisting of 12 DEGs, 6 DELs, and 11 DEMs was constructed, and the top ranked RNAs were Sprouty RTK Signaling Antagonist 2 (Spry2), MSTRG.16386.1, and miR-466b-5p, which were verified using qRT-PCR. We found that MSTRG.16386.1 overexpression resulted in NRK-52E cell apoptosis, which was suppressed by the interaction with miR-466b-5p. In addition, we demonstrated that Spry2 is a key target of MSTRG.16386.1 and miR-466b-5p. Rescue experiment results revealed that the downregulated expression of Spry2 protected NRK-52E cells against apoptosis mediated by MSTRG.16386.1 overexpression or an miR-466b-5p inhibitor. Our findings provide novel insights into ceRNA regulation of the MSTRG.16386.1/miR-466b-5p/Spry2 axis in DHCA-induced kidney injury by the induction of apoptosis.
Assuntos
Injúria Renal Aguda , Parada Circulatória Induzida por Hipotermia Profunda , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas de Membrana , MicroRNAs , RNA Longo não Codificante , Animais , MicroRNAs/genética , MicroRNAs/metabolismo , Ratos , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Masculino , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/genética , Injúria Renal Aguda/patologia , Ratos Sprague-Dawley , Parada Circulatória Induzida por Hipotermia Profunda/efeitos adversos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Transdução de Sinais , Humanos , Apoptose , RNA Endógeno Competitivo , Proteínas do Tecido NervosoRESUMO
OBJECTIVE: The inflammation-responsive NLRP3 inflammasome and nuclear factor-kappa B (NF-κB) dependent signaling pathways are critically connected with inflammatory conditions and disorders such as colorectal cancer (CRC). Where phytochemicals may be added as preventive natural supplements to control CRC. In this study, we investigated whether astilbin (AST) interacted with anti-inflammatory NLRP3 and NF-κB-dependent molecular events in CRC. BACKGROUND: The network of growth signaling redox-sensitive transcription factor NF-κB interacting with the NLRP3 inflammasome in CRC progression needed targeted research. AST is a flavonol reported for anti-inflammatory, immune-suppressive, and antioxidant properties, which are sought to assess CRC growth inhibition through NF-κB and NLRP3. METHODS: AST was applied to HCT116 and HT-29 cells of human origin to examine cell survival, apoptosis, progression, and DNA fragmentation to determine the analysis of gene and protein expression and molecular mechanisms. RESULTS: AST inhibits the proliferation of HCT116 and HT29, and both cell lines depend on IC50 values of 128 and 144 events. AST caused induction of apoptosis in CRC cells via intrinsic mechanism involving caspase-9 and caspase-3 activation and caused arrest of the G2/M phase cell cycle. AST (100 µm) inhibited colony formation abilities to 54% and wound healing to 62% in both cell lines. In the azoxymethane/dextran sodium sulfate (AOM/DSS)-induced colitis-associated colon cancer mice, the total tumor count was reduced from 14 to 4.3 by the AST 20 mg/kg group, significantly reducing the large tumor count. AST (20 mg/kg) suppressed the colonic inflammation as shown by decreased expression of NF-κB (1.8-fold) and NLRP3 (1.5-fold) against the control (1.0-fold). AST restored colon length and histopathological changes caused by AOM/DSS. AST inhibited the production of COX-2, INOS, and pro-inflammatory cytokines and chemokines, especially interleukin-6 (IL-6), IL-1ß, and IL-10, by approximately 50% at 20 mg/kg. AST suppressed intestinal tissue ASC and IL-1ß NLRP3 and NF-kB by approximately 1.3 times compared to control. CONCLUSIONS: AST inhibited colorectal carcinoma growth by blocking the expression of NLRP3 and NF-κB and inducing an apoptotic cascade and suppressing iNOS-COX2 and IL-1ß as regulators of inflammation and growth signal. This study advocates the application of phytopharmaceutical supplements for the management of colorectal carcinoma.
Assuntos
Anti-Inflamatórios , Colite , Neoplasias Colorretais , Flavonóis , Inflamassomos , NF-kappa B , Proteína 3 que Contém Domínio de Pirina da Família NLR , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Humanos , Animais , NF-kappa B/metabolismo , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Neoplasias Colorretais/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Inflamassomos/metabolismo , Flavonóis/farmacologia , Flavonóis/uso terapêutico , Camundongos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Células HT29 , Células HCT116 , Colite/tratamento farmacológico , Colite/patologia , Colite/metabolismo , Colite/induzido quimicamente , Apoptose/efeitos dos fármacos , MasculinoRESUMO
Lung adenocarcinoma (LUAD), a major subtype of non-small-cell lung cancer, exhibits high incidence and mortality rates. Radiotherapy is a critical treatment modality for LUAD, yet its efficacy is often compromised by radiotherapy resistance. (-)-Epicatechin (EC), a natural flavanol derived from Fagopyrum cymosum, has been reported to enhance radiosensitivity in various cancers. However, its specific role and underlying mechanisms in LUAD radiotherapy resistance remain unclear. In this study, we established radiotherapy-resistant cell lines A549R and NCI-H520R by repeatedly irradiating A549 and NCI-H520 cells with gradient doses of X-rays. Furthermore, a xenograft tumor model was constructed by subcutaneously inoculating A549R cells into the left dorsal region of nude mice. The results demonstrated that the forkhead box M1 (FOXM1)-a key oncoprotein implicated in lung cancer proliferation, invasion, and therapy resistance-was significantly upregulated in LUAD tissues and radiotherapy-resistant A549R cells. Knockdown of FOXM1 enhanced radiosensitivity in A549R cells, promoted radiation-induced apoptosis, and suppressed cell proliferation. EC effectively potentiated the radiotherapy response of A549R cells in vitro and attenuated radiotherapy resistance while inhibiting tumor growth in vivo. Mechanistically, EC downregulated ALKBH5, an m6A demethylase known to participate in cancer biology by regulating mRNA demethylation, thereby promoting m6A methylation of FOXM1 mRNA and subsequently suppressing FOXM1 expression, ultimately mitigating radiotherapy resistance in LUAD. This study reveals a novel mechanism by which EC enhances radiosensitivity in LUAD via the ALKBH5/FOXM1 axis, offering a potential therapeutic strategy to overcome radiotherapy resistance in LUAD.
Assuntos
Adenocarcinoma de Pulmão , Homólogo AlkB 5 da RNA Desmetilase , Catequina , Proteína Forkhead Box M1 , Neoplasias Pulmonares , Tolerância a Radiação , Proteína Forkhead Box M1/genética , Proteína Forkhead Box M1/metabolismo , Humanos , Animais , Catequina/farmacologia , Tolerância a Radiação/efeitos dos fármacos , Tolerância a Radiação/genética , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/radioterapia , Adenocarcinoma de Pulmão/metabolismo , Adenocarcinoma de Pulmão/tratamento farmacológico , Adenocarcinoma de Pulmão/patologia , Camundongos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/radioterapia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/tratamento farmacológico , Homólogo AlkB 5 da RNA Desmetilase/genética , Homólogo AlkB 5 da RNA Desmetilase/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Linhagem Celular Tumoral , Camundongos Nus , Ensaios Antitumorais Modelo de Xenoenxerto , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Células A549RESUMO
Multiple myeloma (MM) remains an incurable hematologic malignancy, necessitating novel therapeutic strategies. This study investigates the clinical significance of adiponectin receptors and the anti-myeloma efficacy of their agonist, AdipoRon. Bioinformatic analysis of GEO datasets (GSE124489, GSE187009) revealed significant downregulation of ADIPOR1 and ADIPOR2 in MM patients. Low expression of ADIPOR1 correlated with poor prognosis. Functionally, AdipoRon exerted potent anti-proliferative effects on MM cell lines (U266, RPMI8226) in time- and dose-dependent manners. Mechanistic studies demonstrated that AdipoRon induced mitochondrial apoptosis, evidenced by increased cleavage of PARP and Caspase-9, and triggered G0/G1 cell cycle arrest. At the signaling level, AdipoRon activated the AMPK pathway while concurrently suppressing AKT phosphorylation. The critical role of AMPK was confirmed through pharmacological approaches: the AMPK activator AICAR mimicked AdipoRon's effects, whereas the AMPK inhibitor Compound C partially reversed them. Further investigation identified acetyl-CoA carboxylase (ACC) as a key downstream effector, with ACC inhibition (TOFA) recapitulating AdipoRon's anti-MM effects. Specifically, AdipoRon preferentially suppressed ACC1 expression and subsequently downregulated CPT1A, indicating disruption of fatty acid metabolism. These findings establish that AdipoRon suppresses MM progression through AMPK-driven metabolic reprogramming and apoptosis induction, positioning adiponectin receptor agonism as a promising therapeutic strategy for multiple myeloma.
Assuntos
Proteínas Quinases Ativadas por AMP , Apoptose , Mieloma Múltiplo , Piperidinas , Receptores de Adiponectina , Humanos , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/patologia , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/genética , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Receptores de Adiponectina/genética , Receptores de Adiponectina/metabolismo , Receptores de Adiponectina/agonistas , Proteínas Quinases Ativadas por AMP/metabolismo , Proteínas Quinases Ativadas por AMP/genética , Piperidinas/farmacologia , Acetil-CoA Carboxilase/genética , Acetil-CoA Carboxilase/metabolismo , Acetil-CoA Carboxilase/antagonistas & inibidores , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Linhagem Celular Tumoral , Transdução de Sinais/efeitos dos fármacos , Reprogramação MetabólicaRESUMO
ObjectiveThe study aimed to explore the inhibitory effect of stevioside on colorectal cancer and its molecular mechanism.MethodsColorectal cancer cells were selected for functional testing, including the following groups: 0 µM stevioside, 1 µM stevioside, 2.5 µM stevioside and 5 µM stevioside. CCK-8 kit and EdU staining were employed to assess the cell viability. Cell apoptosis was deterred by flow cytometry. Western blot assay was utilized to detect the protein expressions of cleaved-caspase-3, Bax, Bcl-2, E-cadherin and Vimentin. The polarization of macrophage was evaluated through flow cytometry, western blot and immunofluorescence staining. The effect of stevioside on the proliferation of tumor tissue was detected by tumor formation and immunohistochemical staining in nude mice.ResultsStevioside exhibited a significant concentration-dependent inhibitory effect on the proliferation, migration, and invasion of colorectal cancer cells, while promoting apoptosis in vitro. Following stevioside treatment, there was a notable reduction in tumor volume and weight observed. Flow cytometry and immunohistochemical staining results showed that compared with control group, CD86+ cell ratio was increased in stevioside treatment group, while the CD206+ cell ratio was decreased in stevioside treatment group. RT-qPCR analysis revealed that, compared to the control group, stevioside treatment significantly reduced the mRNA expressions of Arg-1 and IL-10, while concomitantly increasing the mRNA expressions of IL-12 and TNF-α in a concentration-dependent manner.ConclusionStevioside possesses the ability to significantly hinder the proliferation of colorectal cancer cells and induce apoptosis, the mechanism of which may be closely related to the regulation of macrophage M1 polarization.
Assuntos
Neoplasias Colorretais , Diterpenos do Tipo Caurano , Glucosídeos , Macrófagos , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Animais , Humanos , Glucosídeos/farmacologia , Glucosídeos/uso terapêutico , Macrófagos/efeitos dos fármacos , Diterpenos do Tipo Caurano/farmacologia , Diterpenos do Tipo Caurano/uso terapêutico , Camundongos , Camundongos Nus , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Linhagem Celular Tumoral , Camundongos Endogâmicos BALB C , MasculinoRESUMO
PURPOSE: Previous work demonstrated that supraphysiological glucose remodels TGF-ß1 and NF-κB signaling in human limbal stromal cells (LSCs) and congenital aniridia-derived LSCs (AN-LSCs). The present study investigated whether the same metabolic stress also alters apoptotic pathways in these cells. METHODS: Primary LSCs (n = 12) and AN-LSCs (n = 8) were cultured for 48 hours in DMEM containing either normal (17.5 mM) or high (70 mM) glucose. Apoptosis was quantified by Annexin V/propidium iodide (PI) flow cytometry (FC). Expression levels of apoptosis-related genes-including CASP3/7/8/9/10, BCL2, BID, BAX, CDKN1A (p21), CDKN1B (p27), TNFα, XIAP, and BIRC5 (Survivin)-were assessed by qPCR. Protein levels of these markers were analyzed by FC, and TNFα protein concentrations in culture supernatants were measured by ELISA. RESULTS: High glucose significantly reduced the proportion of apoptotic cells in both LSCs (p = 0.0170) and AN-LSCs (p = 0.0181). In both cell types, CASP8 (p = 0.0448; p = 0.0171) and CASP10 (p = 0.0001; p = 0.0007) mRNA levels decreased, while XIAP (p = 0.0375; p = 0.0442) and BIRC5 (p = 0.0196; p = 0.0003) were upregulated. AN-LSCs additionally showed reductions in CASP3 (p = 0.0138) and CDKN1A (p = 0.0331), and exhibited lower BAX levels than LSCs under high glucose (p = 0.0255). Protein analysis corroborated these findings in AN-LSCs: Caspase-3 (p = 0.0154) and Caspase-8 (p = 0.0257) decreased, while Bcl-2 (p = 0.0284) and Survivin (p = 0.0467) levels increased. XIAP protein levels rose in both LSCs (p = 0.0451) and AN-LSCs (p = 0.0134). CONCLUSIONS: A 48-hour exposure to 70 mM glucose induces a marked anti-apoptotic shift in human limbal stromal cells, more pronounced in cells derived from congenital aniridia patients. Together with previous evidence on TGF-ß1 and NF-κB regulation, these findings suggest that limbal cells may mount an early protective response to metabolic stress, which could be harnessed therapeutically to manage aniridia-associated keratopathy through coordinated survival and stress pathways.
Assuntos
Apoptose , Glucose , Limbo da Córnea , Células Estromais , Humanos , Glucose/farmacologia , Apoptose/efeitos dos fármacos , Células Estromais/metabolismo , Células Estromais/efeitos dos fármacos , Células Estromais/patologia , Limbo da Córnea/patologia , Limbo da Córnea/metabolismo , Limbo da Córnea/citologia , Células Cultivadas , Fator de Necrose Tumoral alfa/metabolismo , Feminino , Masculino , Transdução de Sinais/efeitos dos fármacosRESUMO
Colon cancer (CC) is a malignancy with high global incidence and mortality, and elucidating its underlying molecular mechanisms is critical for improving prognostic assessment and therapeutic strategies. In this study, transcriptomic data from a large cohort of CC samples and a limited number of normal controls from the TCGA database were used to construct a multigene prognostic risk model using univariate Cox and least absolute shrinkage and selection operator (LASSO) regression analyses. The expression of key prognostic genes, including immunoglobulin superfamily member 9 (IGSF9), was further validated in CC tissues by PCR and Western blotting. Functional assays were performed in HCT116 cells to investigate the biological effects of IGSF9 overexpression and its regulatory relationship with p53. The prognostic model identified IGSF9 as a gene significantly associated with patient survival. Although IGSF9 expression was reduced at both the mRNA and protein levels in CC tissues, its overexpression in vitro markedly promoted apoptosis, alleviated DNA damage, and suppressed cell migration and invasion. Notably, silencing of p53 partially reversed the tumor-suppressive effects induced by IGSF9 overexpression, indicating that IGSF9 exerts its biological functions in a p53-dependent manner. Collectively, these findings demonstrate that IGSF9 acts as a tumor suppressor in colorectal cancer and regulates DNA damage responses and apoptosis through a mechanism partially dependent on p53, highlighting its potential value as a prognostic biomarker and therapeutic target in CC.
Assuntos
Neoplasias do Colo , Reparo do DNA , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Imunoglobulinas , Proteínas de Membrana , Proteína Supressora de Tumor p53 , Humanos , Proteína Supressora de Tumor p53/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/antagonistas & inibidores , Neoplasias do Colo/patologia , Neoplasias do Colo/metabolismo , Neoplasias do Colo/genética , Células HCT116 , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Feminino , Apoptose , Masculino , Movimento Celular , PrognósticoRESUMO
BACKGROUND: To investigate the mechanism of chronic intermittent hypoxia on gastric injury in rats and the intervening effect and possible mechanism of melatonin. METHODS: Forty-eight male Wistar rats were randomly divided into normal control, intermittent hypoxia, and melatonin treatment groups. Subgroups (n = 4 per time point) were treated for 2, 4, 6, and 8 weeks. Gastric tissue morphology, gastric juice pH, pepsin levels, oxidative stress markers (MDA and SOD), and the expression of JNK and apoptosis-related genes (Bax, Bcl-2) were assessed. RESULTS: The intermittent hypoxia group exhibited significant gastric mucosal damage, decreased pH, increased pepsin, elevated MDA, reduced SOD, and upregulation of JNK and Bax/Bcl-2 mRNA ratio. Melatonin treatment markedly alleviated these pathological and molecular changes compared to the intermittent hypoxia group (P < 0.05). CONCLUSION: Chronic intermittent hypoxia induces gastric mucosal injury, which is associated with oxidative stress imbalance and activation of JNK-mediated apoptotic signaling. Melatonin exerts a protective effect by enhancing antioxidant capacity and suppressing the JNK-Bax/Bcl-2 pathway.
Assuntos
Apoptose , Mucosa Gástrica , Hipóxia , Sistema de Sinalização das MAP Quinases , Melatonina , Estresse Oxidativo , Animais , Melatonina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Masculino , Hipóxia/complicações , Hipóxia/metabolismo , Hipóxia/patologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Mucosa Gástrica/lesões , Ratos , Ratos Wistar , Proteína X Associada a bcl-2/metabolismo , Proteína X Associada a bcl-2/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Antioxidantes/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Superóxido Dismutase/metabolismo , Malondialdeído/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
Immunotherapy has emerged as a promising approach in the management of cancer. However, the suboptimal efficacy of immunotherapy monotherapy underscores the need to develop more effective combination strategies. In this study, we focused on PSMD1 to investigate its role and the molecular pathways by which it regulates the response to immunotherapy in hepatocellular carcinoma (HCC). In HCC, elevated PSMD1 levels are linked to associated with poor prognosis. PSMD1 was predominantly expressed in malignant epithelial cells. Tissue microarray results showed that PSMD1 was highly expressed in tumor tissues. Silencing PSMD1 suppressed HCC cell proliferation and promoted apoptosis in both in vitro and in vivo models. Additionally, PSMD1 suppression decreased PD-L1 expression, thereby enhancing the therapeutic efficacy of anti-PD-1 therapy. Mechanistically, publicly available single-cell RNA sequencing (scRNA-seq) datasets indicated that PSMD1 positively regulates ß-catenin signaling. Silencing of PSMD1 decreased the expression of ß-catenin pathway-associated proteins. Further analysis via mass spectrometry revealed that PSMD1 interacts with Rhotekin (RTKN) and suppresses its ubiquitination. Subsequent experiments revealed that RTKN enhances ß-catenin expression through AKT phosphorylation, thereby increasing PD-L1 transcription. In summary, our findings demonstrate that PSMD1 regulates RTKN protein expression, whereas RTKN facilitates ß-catenin expression via AKT phosphorylation. This mechanism contributes to HCC progression and the effectiveness of immunotherapy. The PSMD1/RTKN/ß-catenin axis could serve as a promising therapeutic target for HCC.