RESUMEN
Cardiovascular disease is a growing major global public health problem. Oxidative stress is regarded as one of the key regulators of pathological physiology, which eventually leads to cardiovascular disease. However, mechanisms by which FGF-2 rescues cells from oxidative stress damage in cardiovascular disease is not fully elucidated. Herein this study was designed to investigate the protective effects of FGF-2 in H2O2-induced apoptosis of H9c2 cardiomyocytes, as well as the possible signaling pathway involved. Apoptosis of H9c2 cardiomyocytes was induced by H2O2 and assessed using methyl thiazolyl tetrazolium assay, Hoechst, and TUNEL staining. Cells were pretreated with PI3K/Akt inhibitor LY294002 to investigate the possible PI3K/Akt pathways involved in the protection of FGF-2. The levels of p-Akt, p-FoxO3a, and Bim were detected by immunoblotting. Stimulation with H2O2 decreased the phosphorylation of Akt and FoxO3a, and induced nuclear localization of FoxO3a and apoptosis of H9c2 cells. These effects of H2O2 were abrogated by pretreatment with FGF-2. Furthermore, the protective effects of FGF-2 were abolished by PI3K/Akt inhibitor LY294002. In conclusion, our data suggest that FGF-2 protects against H2O2-induced apoptosis of H9c2 cardiomyocytes via activation of the PI3K/Akt/FoxO3a pathway.
Asunto(s)
Apoptosis/efectos de los fármacos , Factor 2 de Crecimiento de Fibroblastos/fisiología , Proteína Forkhead Box O3/metabolismo , Peróxido de Hidrógeno/toxicidad , Miocitos Cardíacos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Animales , Proteína 11 Similar a Bcl2/metabolismo , Línea Celular , Fosforilación , Transporte de Proteínas , RatasRESUMEN
Diabetes is known to be associated with neurodegenerative diseases. Resveratrol, a plant-derived polyphenolic compound found in red wine, possesses antioxidant properties. In this study, we aimed to investigate the effects of resveratrol on the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt)/FoxO3a pathway in mediating high glucose (HG)-induced injuries in neuronal PC12 cells. PC12 cells were exposed to HG to establish a model of HG neurotoxicity. Results showed that pre-treating PC12 cells with resveratrol before exposure to HG led to increased cell viability, decreased apoptotic cells, and reactive oxygen species generation. Western blot analysis showed that HG decreased the phosphorylation of Akt and FoxO3a and led to the nuclear localization of FoxO3a. These effects were significantly alleviated by resveratrol co-treatment. Furthermore, the protective effects of resveratrol were abolished by PI3K/Akt inhibitor LY294002. All these results demonstrate that resveratrol protected the PC12 cells from HG-induced oxidative stress and apoptosis via the activation of PI3K/Akt/FoxO3a signaling pathway.
Asunto(s)
Factores de Transcripción Forkhead/metabolismo , Glucosa/toxicidad , Neuroprotección/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Neurotoxinas/toxicidad , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Estilbenos/farmacología , Acetilcisteína/farmacología , Animales , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteína 11 Similar a Bcl2 , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Cromonas/farmacología , Regulación hacia Abajo/efectos de los fármacos , Proteína Forkhead Box O3 , Proteínas de la Membrana/metabolismo , Morfolinas/farmacología , Estrés Oxidativo/efectos de los fármacos , Células PC12 , Fosforilación/efectos de los fármacos , Proteínas Proto-Oncogénicas/metabolismo , Ratas , Especies Reactivas de Oxígeno/metabolismo , Resveratrol , Transducción de Señal/efectos de los fármacosRESUMEN
The sterol regulatory element-binding protein (SREBP) activation and cytokine level were significantly increased in coronavirus disease-19. The NLRP3 inflammasome is an amplifier for cellular inflammation. This study aimed to elucidate the modulatory effect of SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 NP) on trimethylamine N-oxide (TMAO)-induced lipogenesis and NLRP3 inflammasome activation and the underlying mechanisms in vascular smooth muscle cells (VSMCs). Our data indicated that SARS-CoV-2 NP activates the dissociation of the SREBP cleavage activating protein (SCAP) from the endoplasmic reticulum, resulting in SREBP activation, increased lipogenic gene expression, and NLRP3 inflammasome activation. TMAO was applied to VSMC-induced NLRP3 inflammasome by promoting the SCAP-SREBP complex endoplasmic reticulum-to-Golgi translocation, which facilitates directly binding of SARS-CoV-2 NP to the NLRP3 protein for NLRP3 inï¬ammasome assembly. SARS-CoV-2 NP amplified the TMAO-induced lipogenic gene expression and NLRP3 inflammasome. Knockdown of SCAP-SREBP2 can effectively reduce lipogenic gene expression and alleviate NLRP3 inflammasome-mediated systemic inflammation in VSMCs stimulated with TMAO and SARS-CoV-2 NP. These results reveal that SARS-CoV-2 NP amplified TMAO-induced lipogenesis and NLRP3 inflammasome activation via priming the SCAP-SREBP signaling pathway.
Asunto(s)
COVID-19 , Metilaminas , Proteínas de Unión a los Elementos Reguladores de Esteroles , Humanos , Proteínas de Unión a los Elementos Reguladores de Esteroles/metabolismo , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo , SARS-CoV-2 , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Transducción de Señal , Inflamación , Proteínas de la NucleocápsideRESUMEN
Hyperphosphatemia or severe acute respiratory syndrome coronavirus 2 (SARSCoV2) infection can promote cardiovascular adverse events in patients with chronic kidney disease. Hyperphosphatemia is associated with elevated inflammation and sterol regulatory element binding protein 2 (SREBP2) activation, but the underlying mechanisms in SARSCoV2 that are related to cardiovascular disease remain unclear. The present study aimed to elucidate the role of excess inorganic phosphate (PI) in SARSCoV2 N proteininduced NLRP3 inflammasome activation and the underlying mechanisms in vascular smooth muscle cells (VSMCs). The expression levels of SARSCoV2 N protein, SREBP cleavageactivating protein (SCAP), mature Nterminal SREBP2, NLRP3, procaspase1, cleaved caspase1, IL1ß and IL18 were examined by western blotting. The expression levels of SREBP2, HMGCoA reductase, HMGCS1, low density lipoprotein receptor, proprotein convertase subtilisin/kexin type 9 (PCSK9), SREBP1c, fatty acid synthase, stearyl coenzyme A desaturase 1, acetylCoA carboxylase α and ATPcitrate lyase were determined by reverse transcriptionquantitative PCR. The translocation of SCAP or NLRP3 from the endoplasmic reticulum to the Golgi was detected by confocal microscopy. The results showed that excess PI promoted SCAPSREBP and NLRP3 complex translocation to the Golgi, potentially leading to NLRP3 inflammasome activation and lipogenic gene expression. Furthermore, PI amplified SARSCoV2 N proteininduced inflammation via the SCAPSREBP pathway, which facilitates NLRP3 inï¬ammasome assembly and activation. Inhibition of phosphate uptake with phosphonoformate sodium alleviated NLRP3 inflammasome activation and reduced SREBPmediated lipogenic gene expression in VSMCs stimulated with PI and with SARSCoV2 N protein overexpression. Inhibition of SREBP2 or small interfering RNAinduced silencing of SREBP2 effectively suppressed the effect of PI and SARSCoV2 N protein on NLRP3 inflammasome activation and lipogenic gene expression. In conclusion, the present study identified that PI amplified SARSCoV2 N proteininduced NLRP3 inflammasome activation and lipogenic gene expression via the SCAPSREBP signaling pathway.
Asunto(s)
COVID-19 , Hiperfosfatemia , Péptidos y Proteínas de Señalización Intracelular , Proteínas de la Membrana , Humanos , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proproteína Convertasa 9/metabolismo , Proteína 2 de Unión a Elementos Reguladores de Esteroles/genética , Proteína 2 de Unión a Elementos Reguladores de Esteroles/metabolismo , SARS-CoV-2/metabolismo , Fosfatos , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo , Transducción de Señal , InflamaciónRESUMEN
Chronic inflammation is a key factor that accelerates the progression of inflammatory vascular disease. Hydrogen sulfide (H2S) has potent antiinflammatory effects; however, its underlying mechanism of action has not been fully elucidated. The present study aimed to investigate the potential effect of H2S on sirtuin 1 (SIRT1) sulfhydration in trimethylamine Noxide (TMAO)induced macrophage inï¬ammation, and its underlying mechanism. Proinflammatory M1 cytokines (MCP1, IL1ß, and IL6) and antiinflammatory M2 cytokines (IL4 and IL10) were detected by RTqPCR. CSE, p65 NFκB, pp65 NFκB, IL1ß, IL6 and TNFα levels were measured by Western blot. The results revealed that cystathionine γlyase protein expression was negatively associated with TMAOinduced inflammation. Sodium hydrosulfide (a donor of H2S) increased SIRT1 expression and inhibited the expression of inflammatory cytokines in TMAOstimulated macrophages. Furthermore, nicotinamide, a SIRT1 inhibitor, antagonized the protective effect of H2S, which contributed to P65 NFκB phosphorylation and upregulated the expression of inflammatory factors in macrophages. H2S ameliorated TMAOinduced activation of the NFκB signaling pathway via SIRT1 sulfhydration. Moreover, the antagonistic effect of H2S on inflammatory activation was largely eliminated by the desulfhydration reagent dithiothreitol. These results indicated that H2S may prevent TMAOinduced macrophage inï¬ammation by reducing P65 NFκB phosphorylation via the upregulation and sulfhydration of SIRT1, suggesting that H2S may be used to treat inï¬ammatory vascular diseases.
Asunto(s)
Sulfuro de Hidrógeno , Humanos , Cistationina gamma-Liasa/metabolismo , Sulfuro de Hidrógeno/farmacología , Inflamación/metabolismo , Interleucina-6 , Macrófagos/metabolismo , FN-kappa B , Sirtuina 1/metabolismoRESUMEN
Fibroblast growth factor (FGF)21, a member of the family of FGFs, exhibits protective effects against myocardial ischemia and ischemia/reperfusion injury; it is also an enhancer of autophagy. However, the mechanisms underlying the protective role of FGF21 against cardiomyocyte hypoxia/reoxygenation (H/R) injury remain unclear. The present study aimed to investigate the effect of FGF21 on H9c2 cardiomyocyte injury induced by H/R and the mechanism associated with changes in autophagy. Cultured H9c2 cardiomyocytes subjected to hypoxia were treated with a vehicle or FGF21 during reoxygenation. The viability of H9c2 rat cardiomyocytes was measured using Cell Counting Kit8 and trypan blue exclusion assays. The contents of creatine kinase (CK) and creatine kinase isoenzymes (CKMB), cardiac troponin I (cTnT), cardiac troponin T (cTnI) and lactate dehydrogenase (LDH) in culture medium were detected with a CK, CKMB, cTnT, cTnI and LDH assay kits. The protein levels were examined by western blot analysis. Autophagic flux was detected by AdmCherryGFPLC3B autophagy fluorescent adenovirus reagent. The results indicated that FGF21 alleviated H/Rinduced H9c2 myocardial cell injury and enhanced autophagic flux during H/R, and that this effect was antagonized by cotreatment with 3methyladenine, an autophagy inhibitor. Furthermore, FGF21 increased the expression levels of Beclin1 and Vps34 proteins, but not of mechanistic target of rapamycin. These data indicate that FGF21 treatment limited H/R injury in H9c2 cardiomyocytes by promoting autophagic flux through upregulation of the expression levels of Beclin1 and Vps34 proteins.
Asunto(s)
Autofagia , Factores de Crecimiento de Fibroblastos/metabolismo , Hipoxia/metabolismo , Miocitos Cardíacos/metabolismo , Oxígeno/metabolismo , Animales , Biomarcadores , Línea Celular , Supervivencia Celular , Citoprotección , Factores de Crecimiento de Fibroblastos/farmacología , Genes Reporteros , Daño por Reperfusión Miocárdica/etiología , Daño por Reperfusión Miocárdica/metabolismo , Ratas , Transducción de SeñalRESUMEN
Autophagy is a maintenance process for recycling long-lived proteins and cytoplasmic organelles. The level of this process is enhanced during ischemia/reperfusion (I/R) injury. Autophagy can trigger survival signaling in myocardial ischemia, whereas defective autophagy during reperfusion is detrimental. Autophagy can be regulated through multiple signaling pathways in I/R, including Beclin1/class III phosphatidylinositol3 kinase (PI3K), adenosine monophosphate activated protein kinase/mammalian target of rapamycin (mTOR), and PI3K/protein kinase B/mTOR pathways, which consequently lead to different functions. Thus, autophagy has both protective and detrimental functions, which are determined by different signaling pathways and conditions. Targeting the activation of autophagy can be a promising new therapeutic strategy for treating cardiovascular disease.
Asunto(s)
Autofagia , Daño por Reperfusión Miocárdica/fisiopatología , Transducción de Señal , Animales , Humanos , Daño por Reperfusión Miocárdica/metabolismoRESUMEN
Elevated plasma low-density lipoprotein cholesterol (LDL-C) is an important risk factor for cardiovascular diseases. Statins are the most widely used therapy for patients with hyperlipidemia. However, a signiï¬cant residual cardiovascular risk remains in some patients even after maximally tolerated statin therapy. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a new pharmacologically therapeutic target for decreasing LDL-C. PCSK9 reduces LDL intake from circulation by enhancing LDLR degradation and preventing LDLR recirculation to the cell surface. Moreover, PCSK9 inhibitors have been approved for patients with either familial hypercholesterolemia or atherosclerotic cardiovascular disease, who require additional reduction of LDL-C. In addition, PCSK9 inhibition combined with statins has been used as a new approach to help reduce LDL-C levels in patients with either statin intolerance or unattainable LDL goal. This review will discuss the emerging anti-PCSK9 therapies in the regulation of cholesterol metabolism and atherosclerosis.
Asunto(s)
Aterosclerosis/metabolismo , Metabolismo de los Lípidos/fisiología , Proproteína Convertasa 9/metabolismo , Animales , Anticolesterolemiantes/farmacología , Anticolesterolemiantes/uso terapéutico , Aterosclerosis/tratamiento farmacológico , LDL-Colesterol/metabolismo , Humanos , Hipercolesterolemia/tratamiento farmacológico , Hipercolesterolemia/metabolismo , Metabolismo de los Lípidos/efectos de los fármacosRESUMEN
Doxorubicin (DOX) is an efficient drug used in cancer therapy; however, it can induce severe cytotoxicity, which limits its clinical application. In the present study, the effects of resveratrol (RES) on sirtuin 1 (SIRT1) activation in mediating DOX-induced cytotoxicity in H9c2 cardiac cells was investigated. H9c2 cells were exposed to 5 µM DOX for 24 h to establish a model of DOX cardiotoxicity. Apoptosis of H9c2 cardiomyocytes was assessed using the MTT assay and Hoechst nuclear staining. The results demonstrated that pretreating H9c2 cells with RES prior to the exposure of DOX resulted in increased cell viability and a decreased quantity of apoptotic cells. Western blot analysis demonstrated that DOX decreased the expression level of SIRT1. These effects were significantly alleviated by co-treatment with RES. In addition, the results demonstrated that DOX administration amplified forkhead box O1 (FoxO1) and P53 expression levels in H9c2 cells. RES was also found to protect against DOX-induced increases of FoxO1 and P53 expression levels in H9c2 cells. Furthermore, the protective effects of RES were arrested by the SIRT1 inhibitor nicotinamide. In conclusion, the results demonstrated that RES protected H9c2 cells against DOX-induced injuries via SIRT1 activation.
RESUMEN
The hepatocyte growth factor (HGF)/c-Met signaling pathway results in cancer cell scattering and invasion, and has been reported to participate in several types of cancer, including prostate and colorectal cancer. The downstream phosphorylation cascade of HGF, particularly the mitogen-activated protein kinase and phosphoinositide 3-kinase/AKT signaling pathway, regulates epithelial-mesenchymal transition (EMT). However, the mechanism by which these signaling pathways govern EMT, and whether certain kinases are able to respond to specific EMT effectors, remains to be elucidated. In the present study, an increase in the levels of vimentin, rather than co-regulation of certain EMT marker proteins, was observed in response to HGF-induced EMT in DU145 prostate cancer cells. In addition, it was observed that curcumin abrogated HGF-induced DU145 cell scattering and invasion. Furthermore, curcumin was able to effectively inhibit the HGF-induced increase in the levels of vimentin by downregulating the expression of phosphorylated c-Met, extracellular signal-regulated kinase and Snail. In conclusion, the results of the present study demonstrated that curcumin was able to reverse HGF-induced EMT, possibly by inhibiting c-Met expression in DU145 prostate cancer cells.