RESUMO
Neoatherosclerosis is a major cause of stent failure after percutaneous coronary intervention. Metabolism such as hyperuricemia is associated with in-stent restenosis (ISR). However, the association between serum uric acid (sUA) levels and in-stent neoatherosclerosis (ISNA) has never been validated.A total of 216 patients with 220 ISR lesions who had undergone optical coherence tomography (OCT) of culprit stents were included in this study. According to their sUA levels, eligible patients were divided into two groups [normal-sUA group: sUA < 7 mg/dL, n = 126, and high-sUA group: sUA ≥ 7 mg/dL, n = 90]. OCT findings were analyzed and compared between the normal- and high-sUA groups.The incidence of ISNA (63.0% versus 43.0%, P = 0.004) was significantly higher in the high-sUA group than in the normal-sUA group. Lipid plaques (66.3% versus 43.0%, P < 0.001) and thin-cap fibroatheroma (38.0% versus 18.0%, P = 0.001) were observed more frequently in the restenotic tissue structure in patients in the high-sUA group than in those in the normal-sUA group. Meanwhile, univariate (OR: 1.208, 95% CI: 1.037-1.407; P = 0.015) and multivariate (OR: 1.254, 95% CI: 1.048-1.501; P = 0.013) logistic regression analyses indicated that sUA levels were an independent risk factor for ISNA after adjusting for relevant risk factors.The high-sUA levels were an independent risk factor for the occurrence of neoatherosclerosis in patients with ISR via OCT.
Assuntos
Aterosclerose , Reestenose Coronária , Intervenção Coronária Percutânea , Placa Aterosclerótica , Humanos , Ácido Úrico , Reestenose Coronária/etiologia , Reestenose Coronária/complicações , Placa Aterosclerótica/complicações , Stents/efeitos adversos , Aterosclerose/etiologia , Tomografia de Coerência Óptica/métodos , Intervenção Coronária Percutânea/efeitos adversos , Constrição Patológica/patologia , Vasos Coronários/patologiaRESUMO
The Ser/Thr protein phosphatase Ppt1 (yeast)/PP5 (humans) has been implicated in signal transduction-mediated growth and differentiation, DNA damage/repair, cell cycle progression, and heat shock responses. Little, however, is known concerning the functions of Ppt1/PP5 in filamentous fungi. In this study, the Ppt1 gene MaPpt1 was characterized in the insect pathogenic fungus, Metarhizium acridum. The MaPpt1 protein features a three-tandem tetratricopeptide repeat (TPR) domain and a peptidyl-prolyl cis-trans isomerase-like (PP2Ac) domain. Subcellular localization using an MaPpt1::eGFP fusion protein revealed that MaPpt1 was localized in the cytoplasm of spores, but gathered at the septa in growing hyphae. Targeted gene inactivation of MaPpt1 in M. acridum resulted in unexpected reprogramming of normal aerial conidiation to microcycle conidiation. Although overall vegetative growth was unaffected, a significant increase in conidial yield was noted in ΔMaPpt1. Stress-responsive phenotypes and virulence were largely unaffected in ΔMaPpt1. Exceptionally, ΔMaPpt1 displayed increased UV tolerance compared to wild type. Digital gene expression data revealed that MaPpt1 mediates transcription of sets of genes involved in conidiation, polarized growth, cell cycle, cell proliferation, DNA replication and repair, and some important signaling pathways. These data indicate a unique role for Ppt1 in filamentous fungal development and differentiation.
Assuntos
Metarhizium/genética , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , Proliferação de Células/genética , Reparo do DNA/genética , Replicação do DNA/genética , Deleção de Genes , Metarhizium/metabolismo , Transdução de Sinais/genética , Raios UltravioletaRESUMO
2,2',4,4'-tetrabrominated diphenyl ether (BDE-47) and 2,2',4,4',5-pentabromodiphenyl ether (BDE-99) are two typical polybrominated diphenyl ethers (PBDEs), and studies have proven that these PBDs can disrupt the behaviors and physical function of aquatic organisms. However, little is known about the compositional impacts of BDE-47/BDE-99 compound pollution on the feeding behavior of Daphnia magna. In this study, a response surface methodology (RSM) was introduced into the combined toxicity assessment of BDE-47 and BDE-99 on the feeding depression of D. magna. Low concentrations of BDE-47 (9.2⯵g/L) and BDE-99 (5.4⯵g/L) had no effect on the feeding behavior of D. magna; nevertheless, the feeding depression was strengthened, and a concentration-dependent effect was observed with increasing concentrations of BDE-47 and BDE-99. The results of RSM indicated that the mixture of BDE-47 and BDE-99 can enhance their toxicity on the feeding behavior of D. magna. Moreover, real-time PCR (qPCR) analysis showed that the down-regulation of α-amylase (AMS) appeared in most of the exposed D. magna. However, there were significant different in the gene expression of trypsin, superoxide dismutase (SOD) and catalase (CAT) between the exposure and control groups. The change in the enzyme activity of AMS, trypsin, SOD and CAT implied that BDE-47 and BDE-99 cause damage to the digestive and antioxidative systems of D. magna. Correlation analysis indicated that a significant positive correlation existed between the gene expression and enzyme activity of SOD and CAT. Our results contribute to the understanding of toxicity caused by BDE-47/BDE-99 compound pollution in D. magna and help to improve traditional toxicity assessment methods for aquatic environments.
Assuntos
Antioxidantes/metabolismo , Daphnia/efeitos dos fármacos , Digestão/efeitos dos fármacos , Comportamento Alimentar/efeitos dos fármacos , Éteres Difenil Halogenados/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Catalase/genética , Daphnia/enzimologia , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Superóxido Dismutase/genéticaRESUMO
A cobalt-catalyzed C(sp2)-H alkoxylation of 1-naphthylamine derivatives has been disclosed, which represents an efficient approach to synthesize aryl ethers with broad functional group tolerance. It is noteworthy that secondary alcohols, such as hexafluoroisopropanol, isopropanol, isobutanol, and isopentanol, were well tolerated under the current catalytic system. Moreover, a series of biologically relevant fluorine-aryl ethers were easily obtained under mild reaction conditions after the removal of the directing group.
RESUMO
Salicylaldehyde dehydrogenase (sALDH) can oxidize salicylaldehyde, which is an intermediate in the naphthalene catabolism in bacteria. However, genes encoding sALDH have not been discovered so far in Rhodococcus. Here, we report the discovery of a novel aldehyde dehydrogenase (ALDH) gene in the naphthalene degrader Rhodococcus ruber OA1 based on phylogenetic analysis. Interestingly, apart from ALDH activity, ALDH of R. ruber OA1 (OA1-ALDH) also showed sALDH activity. Moreover, its sALDH specific activity was higher than its ALDH specific activity. Based on a comparison with the ALDH of Thermomonospora curvata DSM 43,183, a putative active site Cys123 and NAD+ binding site Asn263 were proposed in R. ruber OA1. Multiple alignment of OA1-ALDH with ALDHs from other organisms indicated that the residues Ser122 and Ala124 might influence the enzyme activity and substrate specificity that render OA1-ALDH the ability to catalyze salicylaldehyde better than acetaldehyde. These results support the possibility that OA1-ALDH plays the role of sALDH in the oxidation of salicylaldehyde to salicylate in R. ruber OA1. In summary, our study would contribute to the understanding of the structure and roles of ALDH in Rhodococcus.
Assuntos
Aldeído Oxirredutases/metabolismo , Aldeídos/metabolismo , Rhodococcus/enzimologia , Domínio Catalítico , Oxirredução , Homologia de Sequência de AminoácidosRESUMO
BACKGROUND: Most fungi, including entomopathogenic fungi, have two different conidiation patterns, normal and microcycle conidiation, under different culture conditions, eg, in media containing different nutrients. However, the mechanisms underlying the conidiation pattern shift are poorly understood. RESULTS: In this study, Metarhizium acridum undergoing microcycle conidiation on sucrose yeast extract agar (SYA) medium shifted to normal conidiation when the medium was supplemented with sucrose, nitrate, or phosphate. By linking changes in nutrients with the conidiation pattern shift and transcriptional changes, we obtained conidiation pattern shift libraries by Solexa/Illumina deep-sequencing technology. A comparative analysis demonstrated that the expression of 137 genes was up-regulated during the shift to normal conidiation, while the expression of 436 genes was up-regulated at the microcycle conidiation stage. A comparison of subtractive libraries revealed that 83, 216, and 168 genes were related to sucrose-induced, nitrate-induced, and phosphate-induced conidiation pattern shifts, respectively. The expression of 217 genes whose expression was specific to microcycle conidiation was further analyzed by the gene expression profiling via multigene concatemers method using mRNA isolated from M. acridum grown on SYA and the four normal conidiation media. The expression of 142 genes was confirmed to be up-regulated on standard SYA medium. Of these 142 genes, 101 encode hypothetical proteins or proteins of unknown function, and only 41 genes encode proteins with putative functions. Of these 41 genes, 18 are related to cell growth, 10 are related to cell proliferation, three are related to the cell cycle, three are related to cell differentiation, two are related to cell wall synthesis, two are related to cell division, and seven have other functions. These results indicate that the conidiation pattern shift in M. acridum mainly results from changes in cell growth and proliferation. CONCLUSIONS: The results indicate that M. acridum shifts conidiation pattern from microcycle conidiation to normal conidiation when there is increased sucrose, nitrate, or phosphate in the medium during microcycle conidiation. The regulation of conidiation patterning is a complex process involving the cell cycle and metabolism of M. acridum. This study provides essential information about the molecular mechanism of the induction of the conidiation pattern shift by single nutrients.
Assuntos
Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Metarhizium/fisiologia , Fenômenos Fisiológicos da Nutrição/genética , Transcriptoma , Biologia Computacional/métodos , Ontologia Genética , Redes e Vias MetabólicasRESUMO
OBJECTIVE: To assess the efficacy and safety of Dan'e-fukang soft extract for dysmenorrhea by meta-analysis. METHODS: Cochrane Controlled Trials Register, PubMed, EMBASE, CBM, VIP, Wanfang Data, and CNKI databases were searched. Results of randomized controlled trials were also harvested from pharmaceutical companies by manual search. Meta-analysis was carried out according to the method provided by the Cochrane Collaboration with RevMan5.0 software. RESULTS: Twelve Chinese papers were selected, and 1213 patients were included. Significant difference in recovery rate was found between Dan'e-fukang soft extract group and other drugs group (RR=1.33, 95%CI: 1.02-1.75, P<0.05), but the difference no longer existed when studies with pseudo ginseng and marvelon were removed from other drug groups (RR=1.08, 95%CI: 0.91-1.29, P>0.05). No statistical difference was noticed in total effective rate between two groups (RR=1.04, 95%CI: 1.00-1.08, P>0.05). A statistical difference in improvement of dysmenorrhea symptoms was found before and after treatment in both Dan'e-fukang soft extract group and other drugs group (MD=5.79, 95%CI: 5.01-6.56, P<0.001; MD=4.62, 95%CI: 3.71-5.53, P<0.001), while no significant difference was seen between two groups before treatment (MD=0.20, 95%CI: -0.11-0.50, P>0.05) and after treatment (MD=-0.94, 95%CI: -2.11-0.23, P>0.05). Oral administration of Dan'e-fukang soft extract caused only mild gastrointestinal discomfort, but other drugs had more adverse effects including serious gastrointestinal reaction, severe liver dysfunction, vaginal bleeding, and female masculinity. CONCLUSION: The existing evidence shows that Dan'e-fukang soft extract has the same efficacy as other drugs in treatment of dysmenorrheal. Because of the quality of the included studies was limited, the evidence of the efficacy and safety of Dan'e-fukang soft extract was not strong, and high-quality randomized trials with large samples are needed.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Dismenorreia/tratamento farmacológico , Feminino , Humanos , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
BACKGROUND: Non-small-cell lung cancer (NSCLC) is a common malignancy with high morbidity and mortality. Circular RNAs are widely involved in NSCLC progression. However, the mechanism of circSLC25A16 in NSCLC has not been reported. METHODS: The expressions of circSLC25A16, microRNA-335-5p (miR-335-5p), and CDGSH iron-sulfur domain-containing protein 2 (CISD2) were monitored by quantitative real-time fluorescence polymerase chain reaction. Western blot was also carried out to measure the protein levels of CISD2, hexokinase 2 (HK2), and lactate dehydrogenase A (LDHA). For functional analysis, cell counting kit-8 assay, 5-ethynyl-2'-deoxyuridine, flow cytometry, transwell, and wound healing assays were utilized to examine cell proliferation, apoptosis, and migration. Glucose uptake and lactate production were detected using commercial kits. The relationship between miR-335-5p and circSLC25A16 or CISD2 was verified by dual-luciferase reporter and RNA immunoprecipitation assays. Furthermore, tumor xenograft was established to explore the function of circSLC25A16 in vivo. RESULTS: CircSLC25A16 and CISD2 were overexpressed in NSCLC, but miR-335-5p was downregulated. CircSLC25A16 acted as a miR-335-5p sponge, and silencing of circSLC25A16 arrested cell proliferation, migration, and glycolysis, and promoted apoptosis, but these impacts were resumed by miR-335-5p inhibition. CISD2 was a miR-335-5p target, and overexpression of CISD2 abolished the suppressive function of miR-335-5p mimic on the malignant behavior of NSCLC cells. CircSLC25A16 could adsorb miR-335-5p to mediate CISD2 expression. Additionally, silencing circSLC25A16 restrained the growth of NSCLC tumor xenograft in vivo. CONCLUSION: CircSLC25A16 facilitated NSCLC progression via the miR-335-5p/CISD2 axis, implying that circSLC25A16 may serve as a novel biomarker for NSCLC treatment.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Proliferação de Células , Neoplasias Pulmonares , MicroRNAs , RNA Circular , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Humanos , MicroRNAs/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/metabolismo , Camundongos , Animais , RNA Circular/genética , Apoptose , Movimento Celular , Camundongos Nus , Regulação Neoplásica da Expressão Gênica , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
ABSTRACT: Cardiac macrophages with different polarization phenotypes regulate ventricular remodeling and neovascularization after myocardial infarction (MI). Annexin A2 (ANXA2) promotes macrophage polarization to the repair phenotype and regulates neovascularization. However, whether ANXA2 plays any role in post-MI remodeling and its underlying mechanism remains obscure. In this study, we observed that expression levels of ANXA2 were dynamically altered in mouse hearts upon MI and peaked on the second day post-MI. Using adeno-associated virus vector-mediated overexpression or silencing of ANXA2 in the heart, we also found that elevation of ANXA2 in the infarcted myocardium significantly improved cardiac function, reduced cardiac fibrosis, and promoted peri-infarct angiogenesis, compared with controls. By contrast, reduction of cardiac ANXA2 exhibited opposite effects. Furthermore, using in vitro coculture system, we found that ANXA2-engineered macrophages promoted cardiac microvascular endothelial cell (CMEC) proliferation, migration, and neovascularization. Mechanistically, we identified that ANXA2 interacted with yes-associated protein (YAP) in macrophages and skewed them toward pro-angiogenic phenotype by inhibiting YAP activity. In addition, ANXA2 directly interacted with integrin ß3 in CMECs and enhanced their growth, migration, and tubule formation. Our results indicate that increased expression of ANXA2 could confer protection against MI-induced injury by promoting neovascularization in the infarcted area, partly through the inhibition of YAP in macrophages and activation of integrin ß3 in endothelial cells. Our study provides new therapeutic strategies for the treatment of MI injury.
Assuntos
Anexina A2 , Traumatismos Cardíacos , Infarto do Miocárdio , Animais , Camundongos , Anexina A2/genética , Anexina A2/metabolismo , Células Endoteliais/metabolismo , Traumatismos Cardíacos/metabolismo , Integrina beta3 , Macrófagos/metabolismo , Infarto do Miocárdio/metabolismo , Miocárdio/metabolismoRESUMO
M2 macrophage-mediated tissue repair plays an important role in acute myocardial infarction (AMI). Additionally, VSIG4, which is mainly expressed on tissue-resident and M2 macrophages, is crucial for the regulation of immune homeostasis; however, its effects on AMI remain unknown. In this study, we aimed to investigate the functional significance of VSIG4 in AMI using VSIG4 knockout and adoptive bone marrow transfer chimeric models. We also determined the function of cardiac fibroblasts (CFs) through gain- or loss-of-function experiments. We showed that VSIG4 promotes scar formation and orchestrates the myocardial inflammatory response after AMI, while also promoting TGF-ß1 and IL-10. Moreover, we revealed that hypoxia promotes VSIG4 expression in cultured bone marrow M2 macrophages, ultimately leading to the conversion of CFs to myofibroblasts. Our results reveal a crucial role for VSIG4 in the process of AMI in mice and provide a potential immunomodulatory therapeutic avenue for fibrosis repair after AMI.
Assuntos
Infarto do Miocárdio , Animais , Camundongos , Fibrose , Hipóxia/metabolismo , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Infarto do Miocárdio/metabolismo , Miocárdio/patologiaRESUMO
Neoatherosclerosis (NA) is a significant contributor to late stent failure; however, predictors of late in-stent restenosis (ISR) with NA have not been systematically reported. This study aimed to identify predictors of NA incidence and plaque vulnerability in patients with late ISR and the role of low-density lipoprotein cholesterol (LDL-C) levels in this process. A total of 216 patients with 216 lesions who underwent optical coherence tomography (OCT) before interventional procedure for late drug-eluting stent ISR were enrolled and divided into NA and non-NA groups based on OCT findings. Results showed that higher LDL-C levels were associated with NA, thin-cap fibroatheroma (TCFA), intimal disruption, plaque erosion, and thrombosis. Multivariate regression analysis revealed that the LDL-C level was an independent risk factor for NA and TCFA. The LDL-C levels exhibited a significant predictive value for NA and TCFA, surpassing other factors such as stent age and other lipid types. In conclusion, a high LDL-C level is an independent predictor of NA incidence and plaque vulnerability in patients with late ISR.
Assuntos
Aterosclerose , Doença da Artéria Coronariana , Reestenose Coronária , Stents Farmacológicos , Doenças das Valvas Cardíacas , Placa Aterosclerótica , Humanos , Placa Aterosclerótica/patologia , Stents Farmacológicos/efeitos adversos , LDL-Colesterol , Tomografia de Coerência Óptica/métodos , Neointima , Valor Preditivo dos Testes , Reestenose Coronária/diagnóstico por imagem , Reestenose Coronária/etiologia , Reestenose Coronária/patologia , Aterosclerose/patologia , Constrição Patológica/complicações , Doenças das Valvas Cardíacas/complicações , Vasos Coronários/diagnóstico por imagem , Vasos Coronários/patologia , Doença da Artéria Coronariana/diagnóstico por imagem , Doença da Artéria Coronariana/terapia , Doença da Artéria Coronariana/complicaçõesRESUMO
PTEN (phosphatase and tensin homologue) is a phosphatase that dephosphorylates both protein and phosphoinositide substrates. It is mutated in a variety of human tumours and has important roles in a diverse range of biological processes, including cell migration and chemotaxis. PTEN's intracellular localization and presumably activity are regulated by chemoattractants in Dictyostelium and mouse neutrophils. However, the mechanisms for its regulation remain elusive. Here we show that RhoA and Cdc42, members of the Rho family of small GTPases, regulate the intracellular localization of PTEN in leukocytes and human transfected embryonic kidney cells. In addition, active RhoA is able to stimulate the phospholipid phosphatase activity of PTEN in human embryonic kidney cells and leukocytes, and this regulation seems to require RhoA's downstream effector, RhoA-associated kinase (Rock). Furthermore, we have identified key residues on PTEN that are required for its regulation by the small GTPase, and show that small GTPase-mediated regulation of PTEN has a significant role in the regulation of chemotaxis.
Assuntos
Monoéster Fosfórico Hidrolases/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Proteína cdc42 de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Animais , Quimiotaxia/fisiologia , Dictyostelium/enzimologia , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Leucócitos/metabolismo , Camundongos , Modelos Biológicos , Neutrófilos/metabolismo , PTEN Fosfo-Hidrolase , Proteínas Serina-Treonina Quinases/metabolismo , Transporte Proteico/fisiologia , Quinases Associadas a rhoRESUMO
BACKGROUND: It is sometimes necessary for the laboratory to re-test samples for critical serum electrolyte levels. It is important to assure reproducibility of results when testing is performed on stored, refrigerated samples. We have tested the reproducibility of results for the critical electrolytes, Na, K, Cl and Ca, from ten randomly selected patients'sera over our maximum storage period of nine (9) days on the Siemens Advia 1800 analyzer. The ranges for each electrolyte were 131-150 meq/L (Na), 3.4-5.2 meq/L (K), 101-123 meq/L (Cl) and 7.3-9.9 mg/dL (Ca). METHODS: We used ion-selective electrodes for Na, K and Cl and the ortho-cresolphthalein dye method for Ca. RESULTS: We find that the reproducibility of determinations for all of these electrolytes was excellent, i.e. the coefficients of variation for each electrolyte determination for each patient were low. CONCLUSION: The methods of measurement for these electrolytes on the Advia 1800 are reliable and reproducible.
Assuntos
Análise Química do Sangue/instrumentação , Análise Química do Sangue/métodos , Cloretos/sangue , Eletrólitos/sangue , Sódio/sangue , Análise Química do Sangue/normas , Cálcio/sangue , Humanos , Eletrodos Seletivos de Íons , Potássio/sangue , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To observe the effects of Di' ao Xinxuekang Soft Capsule (DK) on the plasma levels of superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), and endothelin (ET) in patients with coronary heart disease (CHD), and to study its underlying mechanisms. METHODS: Totally 100 patients with CHD confirmed by coronary angiography were randomly assigned to two groups, the control group (60 cases) and the DK treatment group (40 cases). Patients in the control group received conventional therapy, while those in the DK treatment group received DK additionally. The therapeutic course for all was 3 months. The plasma levels of SOD, MDA, ET, and NO were determined pre-treatment, 4, 8, and 12 weeks after treatment. RESULTS: Compared with before treatment, the serum levels of SOD and NO increased and the levels of MDA and ET decreased at each time point. Besides, better effects were obtained in the DK treatment group (P < 0.05). CONCLUSION: DK possibly played a role in inhibiting lipid peroxidation and improving the endothelial dysfunction.
Assuntos
Doença das Coronárias/tratamento farmacológico , Doença das Coronárias/metabolismo , Medicamentos de Ervas Chinesas/uso terapêutico , Endotélio Vascular/fisiopatologia , Peroxidação de Lipídeos , Idoso , Endotelinas/sangue , Feminino , Humanos , Masculino , Malondialdeído/sangue , Pessoa de Meia-Idade , Óxido Nítrico/sangue , Fitoterapia , Superóxido Dismutase/sangueRESUMO
Atherosclerosis is an inflammatory disease that is associated with monocyte recruitment and subsequent differentiation into lipid-laden macrophages at sites of arterial lesions, leading to the development of atherosclerotic plaques. PLC is a key member of signaling pathways initiated by G protein-coupled ligands in macrophages. However, the role of this enzyme in the regulation of macrophage function is not known. Here, we studied macrophages from mice lacking PLC beta2, PLC beta3, or both PLC isoforms and found that PLC beta3 is the major functional PLC beta isoform in murine macrophages. Although PLC beta3 deficiency did not affect macrophage migration, adhesion, or phagocytosis, it resulted in macrophage hypersensitivity to multiple inducers of apoptosis. PLC beta3 appeared to regulate this sensitivity via PKC-dependent upregulation of Bcl-XL. The significance of PLC beta signaling in vivo was examined using the apoE-deficient mouse model of atherosclerosis. Mice lacking both PLC beta3 and apoE exhibited fewer total macrophages and increased macrophage apoptosis in atherosclerotic lesions, as well as reduced atherosclerotic lesion size when compared with mice lacking only apoE. These results demonstrate what we believe to be a novel role for PLC activity in promoting macrophage survival in atherosclerotic plaques and identify PLC beta3 as a potential target for treatment of atherosclerosis.
Assuntos
Apoptose , Aterosclerose/enzimologia , Macrófagos Peritoneais/enzimologia , Fosfolipase C beta/metabolismo , Transdução de Sinais , Animais , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Apoptose/genética , Aterosclerose/genética , Aterosclerose/patologia , Adesão Celular/genética , Linhagem Celular , Movimento Celular/genética , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Inflamação/enzimologia , Inflamação/genética , Inflamação/patologia , Isoenzimas/genética , Isoenzimas/metabolismo , Macrófagos Peritoneais/patologia , Camundongos , Camundongos Knockout , Fagocitose/genética , Fosfolipase C beta/genética , Proteína Quinase C/genética , Proteína Quinase C/metabolismo , Transdução de Sinais/genética , Proteína bcl-X/biossíntese , Proteína bcl-X/genéticaRESUMO
OBJECTIVE: Although earlier studies have shown that the transplantation of mesenchymal stem cells (MSCs) might improve cardiac functions after myocardial infraction, its role on vascular restenosis after percutaneous coronary intervention (PCI) remains controversial. The aim of this study was to investigate the effects of MSCs on the restenosis of injured artery following balloon angioplasty in a rabbit model with both myocardial infarction reperfusion and atherosclerotic stenosis carotid artery by balloon injury. METHODS: After the animal model was established for myocardial infraction reperfusion and atherosclerotic stenosis carotid artery by balloon injury, the rabbits received an intravenous transplantation of MSCs. And an equal volume of phosphate buffered solution was administered for the control group. The animal vascular tissue and myocardium tissue were excised at different time points post-transplantation and used to detect the homing of MSCs and the expressions of platelet-endothelial cell adhesion molecule-1 (CD31) and proliferating cell nuclear antigen (PCNA) by immunohistochemical staining. Four weeks later, vascular restenosis was analyzed by angiography of bilateral carotid arteries and the vascular tissues were used for histological studies. RESULTS: At one week post-transplantation, the 4',6-diamidino-2-phenylindole (DAPI)-labeled MSCs could be detected in myocardial infarction and injured intima. And the intimal expression of CD31 was observed at 2 weeks in the MSCs transplantation group. Yet the expression of PCNA was significantly lower in the MSCs transplantation group than that in the control group (50.5% ± 3.6% vs 23.4% ± 2.8%, P < 0.05). At 4 week post-transplantation, the neointimal area of injured vessels and the vascular restenosis were significantly lower in the MSCs transplantation group than those in the control group (0.092 ± 0.009 vs 0.189 ± 0.007, P < 0.05; 41.7 ± 3.7 vs 61.3 ± 1.6, P < 0.05). Furthermore the MSCs transplantation group demonstrated improved cardiac functions, reduced myocardial infarct size (21.7% ± 2.2% vs 34.3% ± 1.8%, P < 0.05) and significantly increased capillary density around infarction foci (33.6% ± 2.1% vs 20.8% ± 2.6%, P < 0.05) versus the control group. CONCLUSION: The transplantation of MSCs plays significant roles in cardiac repairing in terms of improved cardiac functions, accelerated repair of injured vessels, suppression of neointimal hyperplasia and reduced restenosis of injured vessels.
Assuntos
Transplante de Células-Tronco Mesenquimais , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Animais , Reestenose Coronária/etiologia , Reestenose Coronária/patologia , Modelos Animais de Doenças , Endotélio Vascular/patologia , Infarto do Miocárdio/cirurgia , CoelhosRESUMO
Objective: This study aimed to explore the role of circular RNAs (circRNAs) in M2 macrophage (M2M)-derived small extracellular vesicles (SEVs) in myocardial fibrosis development. Methods: The regulatory role of M2M-derived extracellular vesicles (EVs) was evaluated in a mouse model of acute myocardial infarction. Immunofluorescence, quantitative real-time PCR (RT-qPCR), nanoparticle tracking analysis, Western blot analysis and electron microscopy were used to identify macrophages, large extracellular vesicles (LEVs) and SEVs. The circRNA expression profiles of M0 macrophages (M0Ms) and M2Ms were determined by microarray analysis. Bioinformatic analysis, cell coculture and cell proliferation assays were performed to investigate the expression, function, and regulatory mechanisms of circUbe3a in vitro. qPCR, RNA immunoprecipitation (RIP), dual-luciferase reporter assays, RNA fluorescence in situ hybridization (RNA-FISH), Western blot analysis and a series of rescue experiments were used to verify the correlation among circUbe3a, miR-138-5p and RhoC. Results: CircUbe3a from M2M-derived SEVs triggered functional changes in cardiac fibroblasts (CFs). CircUbe3a was synthesized and loaded into SEVs during increased M2M infiltration after myocardial infarction. The fusion of the released SEVs with the plasma membrane likely caused the release of circUbe3a into the cytosol of CFs. Silencing or overexpressing circUbe3a altered CF proliferation, migration, and phenotypic transformation in vitro. We confirmed that circUbe3a plays a crucial role in enhancing functional changes in CFs by sponging miR-138-5p and then translationally repressing RhoC in vitro. In vivo, the addition of M2M-derived SEVs or overexpression of circUbe3a significantly exacerbated myocardial fibrosis after acute myocardial infarction, and these effects were partially abolished by circUbe3a-specific shRNA. Conclusions: Our findings suggest that M2M-derived circUbe3a-containing SEVs promote the proliferation, migration, and phenotypic transformation of CFs by directly targeting the miR-138-5p/RhoC axis, which may also exacerbate myocardial fibrosis after acute myocardial infarction.
Assuntos
Vesículas Extracelulares/química , Macrófagos/metabolismo , Infarto do Miocárdio/patologia , Miocárdio/patologia , RNA Circular/genética , Animais , Divisão Celular , Movimento Celular , Fibroblastos/metabolismo , Fibrose , Humanos , Camundongos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Infarto do Miocárdio/genética , Miocárdio/metabolismo , Interferência de RNA , RNA Interferente Pequeno/genética , Remodelação Ventricular , Proteína de Ligação a GTP rhoC/fisiologiaRESUMO
Accurate risk assessment of high-risk patients is essential in clinical practice. However, there is no practical method to predict or monitor the prognosis of patients with ST-segment elevation myocardial infarction (STEMI) complicated by hyperuricemia. We aimed to evaluate the performance of different machine learning models for the prediction of 1-year mortality in STEMI patients with hyperuricemia. We compared five machine learning models (logistic regression, k-nearest neighbor, CatBoost, random forest, and XGBoost) with the traditional global (GRACE) risk score for acute coronary event registrations. We registered patients aged >18 years diagnosed with STEMI and hyperuricemia at the Affiliated Hospital of Zunyi Medical University between January 2016 and January 2020. Overall, 656 patients were enrolled (average age, 62.5 ± 13.6 years; 83.6%, male). All patients underwent emergency percutaneous coronary intervention. We evaluated the performance of five machine learning classifiers and the GRACE risk model in predicting 1-year mortality. The area under the curve (AUC) of the six models, including the GRACE risk model, ranged from 0.75 to 0.88. Among all the models, CatBoost had the highest predictive accuracy (0.89), AUC (0.87), precision (0.84), and F1 value (0.44). After hybrid sampling technique optimization, CatBoost had the highest accuracy (0.96), AUC (0.99), precision (0.95), and F1 value (0.97). Machine learning algorithms, especially the CatBoost model, can accurately predict the mortality associated with STEMI complicated by hyperuricemia after a 1-year follow-up.
Assuntos
Hiperuricemia/complicações , Aprendizado de Máquina , Infarto do Miocárdio com Supradesnível do Segmento ST/complicações , Infarto do Miocárdio com Supradesnível do Segmento ST/mortalidade , Idoso , Algoritmos , Área Sob a Curva , China/epidemiologia , Biologia Computacional , Feminino , Fatores de Risco de Doenças Cardíacas , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Intervenção Coronária Percutânea , Prognóstico , Estudos Retrospectivos , Infarto do Miocárdio com Supradesnível do Segmento ST/cirurgiaRESUMO
BACKGROUND: The in-hospital mortality of patients with ST-segment elevation myocardial infarction (STEMI) increases to more than 50% following a cardiogenic shock (CS) event. This study highlights the need to consider the risk of delayed calculation in developing in-hospital CS risk models. This report compared the performances of multiple machine learning models and established a late-CS risk nomogram for STEMI patients. METHODS: This study used logistic regression (LR) models, least absolute shrinkage and selection operator (LASSO), support vector regression (SVM), and tree-based ensemble machine learning models [light gradient boosting machine (LightGBM) and extreme gradient boosting (XGBoost)] to predict CS risk in STEMI patients. The models were developed based on 1,598 and 684 STEMI patients in the training and test datasets, respectively. The models were compared based on accuracy, the area under the curve (AUC), recall, precision, and Gini score, and the optimal model was used to develop a late CS risk nomogram. Discrimination, calibration, and the clinical usefulness of the predictive model were assessed using C-index, calibration plotd, and decision curve analyses. RESULTS: A total of 2282 STEMI patients recruited between January 1, 2016 and May 31, 2020, were included in the complete dataset. The linear models built using LASSO and LR showed the highest overall predictive power, with an average accuracy over 0.93 and an AUC above 0.82. With a C-index of 0.811 [95% confidence interval (CI): 0.769-0.853], the LASSO nomogram showed good differentiation and proper calibration. In internal validation tests, a high C-index value of 0.821 was achieved. Decision curve analysis (DCA) and clinical impact curve (CIC) examination showed that compared with the previous score-based models, the LASSO model showed superior clinical relevance. CONCLUSIONS: In this study, five machine learning methods were developed for in-hospital CS prediction. The LASSO model showed the best predictive performance. This nomogram could provide an accurate prognostic prediction for CS risk in patients with STEMI.
RESUMO
There are a large number of Rho guanine nucleotide exchange factors, most of which have no known functions. Here, we carried out a short hairpin RNA-based functional screen of Rho-GEFs for their roles in leukocyte chemotaxis and identified Arhgef5 as an important factor in chemotaxis of a macrophage phage-like RAW264.7 cell line. Arhgef5 can strongly activate RhoA and RhoB and weakly RhoC and RhoG, but not Rac1, RhoQ, RhoD, or RhoV, in transfected human embryonic kidney 293 cells. In addition, Gbetagamma interacts with Arhgef5 and can stimulate Arhgef5-mediated activation of RhoA in an in vitro assay. In vivo roles of Arhgef5 were investigated using an Arhgef-5-null mouse line. Arhgef5 deficiency did not affect chemotaxis of mouse macrophages, T and B lymphocytes, and bone marrow-derived mature dendritic cells (DC), but it abrogated MIP1alpha-induced chemotaxis of immature DCs and impaired migration of DCs from the skin to lymph node. In addition, Arhgef5 deficiency attenuated allergic airway inflammation. Therefore, this study provides new insights into signaling mechanisms for DC migration regulation.