RESUMO
Excessive inflammatory injury is the main cause of the incidence of severe neonatal pneumonia (NP) and associated deaths. Although dickkopf-3 (DKK3) exhibits anti-inflammatory activity in numerous pathological processes, its role in NP is still unknown. In this study, human embryonic lung WI-38 and MRC-5 cells were treated with lipopolysaccharide (LPS) to induce inflammatory injury of NP in vitro. The expression of DKK3 was downregulated in LPS-stimulated WI-38 and MRC-5 cells. DKK3 overexpression decreased LPS-induced inhibition of cell viability, and reduced LPS-induced apoptosis of WI-38 and MRC-5 cells. DKK3 overexpression also reduced LPS-induced production of pro-inflammatory factors such as ROS, IL-6, MCP-1, and TNF-α. Nuclear respiratory factors 1 (NRF1) knockdown was found to upregulate DKK3 and inactivate the GSK-3ß/ß-catenin pathway in LPS-injured WI-38 and MRC-5 cells. NRF1 knockdown also suppressed LPS-induced inhibition on cell viability, repressed LPS-induced apoptosis, and inhibited the accumulation of ROS, IL-6, MCP-1, and TNF-α in LPS-injured WI-38 and MRC-5 cells. DKK3 knockdown or re-activation of the GSK-3ß/ß-catenin pathway reversed the inhibitory effects of NRF1 knockdown on LPS-induced inflammatory injury. In conclusion, NRF1 knockdown can alleviate LPS-triggered inflammatory injury by regulating DKK3 and the GSK-3ß/ß-catenin pathway.
Assuntos
Pneumonia , Transdução de Sinais , Recém-Nascido , Humanos , Lipopolissacarídeos , Glicogênio Sintase Quinase 3 beta/genética , Glicogênio Sintase Quinase 3 beta/metabolismo , Glicogênio Sintase Quinase 3 beta/farmacologia , beta Catenina/genética , beta Catenina/metabolismo , beta Catenina/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Interleucina-6/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Apoptose/genética , Proteínas Adaptadoras de Transdução de Sinal/genéticaRESUMO
OBJECTIVE: To predict the influencing factors of neonatal pneumonia in pregnant women with diabetes mellitus using a Bayesian network model. By examining the intricate network connections between the numerous variables given by Bayesian networks (BN), this study aims to compare the prediction effect of the Bayesian network model and to analyze the influencing factors directly associated to neonatal pneumonia. METHOD: Through the structure learning algorithms of BN, Naive Bayesian (NB), Tree Augmented Naive Bayes (TAN), and k-Dependence Bayesian Classifier (KDB), complex networks connecting variables were presented and their predictive abilities were tested. The BN model and three machine learning models computed using the R bnlean package were also compared in the data set. RESULTS: In constraint-based algorithms, three algorithms had different presentation DAGs. KDB had a better prediction effect than NB and TAN, and it achieved higher AUC compared with TAN. Among three machine learning modes, Support Vector Machine showed a accuracy rate of 91.04% and 67.88% of precision, which was lower than TAN (92.70%; 72.10%). CONCLUSION: KDB was applicable, and it can detect the dependencies between variables, identify more potential associations and track changes between variables and outcome.
Assuntos
Diabetes Mellitus , Gestantes , Gravidez , Recém-Nascido , Feminino , Humanos , Teorema de Bayes , Algoritmos , Aprendizado de MáquinaRESUMO
BACKGROUND: The primary objective of the study was to determine the diagnostic value of serum N-terminal probrain natriuretic peptide (NT-proBNP) levels to differentiate neonatal pneumonia (NP) and transient tachypnea of the newborn (TTN). The secondary objective was to investigate the prognostic role of NT-proBNP levels in neonates with severe respiratory distress (RD). METHODS: A prospective, observational, single-blinded study involving 58 late preterm and term newborns who were diagnosed with TTN or NP was conducted between June 2020 and June 2021 at a level-3 neonatal intensive care unit in Kirikkale University Faculty of Medicine. TTN and NP groups were compared for serum NT-proBNP levels measured at the 1st and 24th hours of life. Optimal cut-off NT-proBNP value was determined by Youden index to predict the diagnosis of NP. Lung ultrasound was used to support the diagnosis of TTN and NP. In addition, lung ultrasound score (LUS) was used to determine severe RD. RESULTS: The median of NT-proBNP level was significantly higher at the 24th hour of life in the NP group than in the TTN group, respectively 7263.5 pg/mL (1643-35,000) and 3308 pg/mL (69-19,746), p = 0.004. At a cut-off value of 5515.5 pg/mL, NT-proBNP had a sensitivity of 75% and specificity of 73.8% to predict NP [AUC= 0.749 (95% CI: 0.602-0.895; p = 0.004)]. The study population was divided into two groups as high score group (n: 23, LUS ≥ 7) and low score group (n: 35, LUS < 7) according to the LUS at the 6th hour of life. NT-proBNP values at 24th hour of life were 6320 pg/mL (69-35,000) in high score group and 3500 pg/mL (570-15,948) in low score group, p = 0.044. Duration of oxygen support (p = 0.006), noninvasive ventilation (p = 0.008) and NICU stay (p = 0.004) were higher in high-score group. DISCUSSION: NT-proBNP values at 24th hour of life can be used as a relatively early predictor in the differentiation between NP and TTN in late preterm and term neonates. In addition, elevated NT-proBNP values are related to the higher LUS which reflects the severity of RD regardless of diagnosis.
Assuntos
Pneumonia , Taquipneia Transitória do Recém-Nascido , Humanos , Recém-Nascido , Biomarcadores , Peptídeo Natriurético Encefálico , Fragmentos de Peptídeos , Estudos Prospectivos , Taquipneia Transitória do Recém-Nascido/diagnósticoRESUMO
BACKGROUND: Neonatal pneumonia is a common illness in the neonatal period with a high fatality rate. Accumulating proofs have attested to the crucial role of circular RNAs (circRNAs) in pneumonia. This study was intended to expound on the function of circ_0038467 and the underlying mechanism in lipopolysaccharide (LPS)-stimulated 16HBE cell injury in neonatal pneumonia. METHODS: 16HBE cells were exposed to LPS to establish an in vitro neonatal pneumonia cell model. Quantitative real-time polymerase chain reaction (qRT-PCR) was implemented for detecting the levels of circ_0038467, microRNA-545-3p (miR-545-3p), and tumor necrosis factor receptor-associated factor 1 (TRAF1) in neonatal pneumonia serums and LPS-treated 16HBE cells. Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) incorporation, and flow cytometry assays were used to examine cell viability, proliferation, and apoptosis, respectively. The protein abundances of proliferation/apoptosis/inflammation-correlated makers and TRAF1 were tested by Western blot. RNase R and Actinomycin D assays were implemented to determine the features of circ_0038467. The mutual effect between miR-545-3p and circ_0038467 or TRAF1 was affirmed by a dual-luciferase reporter and RNA pull-down assay assays. RESULTS: Circ_0038467 was upregulated in neonatal pneumonia serum specimens and LPS-triggered 16HBE cells. LPS administration restrained 16HBE cell proliferation and promoted apoptosis and inflammation, whereas circ_0038467 silence recovered these influences. Meanwhile, miR-545-3p was targeted by circ_0038467, and circ_0038467 could modulate LPS-treated 16HBE cell injury through absorbing miR-545-3p. Furthermore, circ_0038467 controlled TRAF1 level via segregating miR-545-3p. Moreover, TRAF1 overexpression relieved the suppressive impact of circ_0038467 silence in LPS-triggered 16HBE cell detriment. CONCLUSION: Circ_0038467 knockdown mitigated LPS-exposed 16HBE cell damage through regulating miR-545-3p/PPARA axis.
Assuntos
MicroRNAs , Pneumonia , RNA Circular , Fator 1 Associado a Receptor de TNF , Humanos , Recém-Nascido , Apoptose , Proliferação de Células , Inflamação , Lipopolissacarídeos , MicroRNAs/genética , Fator 1 Associado a Receptor de TNF/genética , RNA Circular/genéticaRESUMO
Neonatal pneumonia is a high neonatal mortality disease. We studied the function and mechanism of long noncoding RNA myocardial infarction-associated transcript 2 (lncRNA MIAT2) on lipopolysaccharide (LPS)-induced inflammation in WI-38 cells. Cell Counting Kit-8 and apoptosis assay were respectively used to detect the functions of LPS, MIAT2, and microRNA-15 (miR-15) on viability and apoptosis. MIAT2 and miR-15 expressions were changed by cell transfection. Moreover, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blot, and enzyme-linked immunosorbent assay were used to detect the expressions of interleukin (IL)-6 and monocyte chemoattractant protein-1 (MCP-1). The levels of Bax, cleaved-caspase-3, and cell pathways-related proteins were tested by western blot. Besides, the levels of miR-15 and MIAT2 were tested by RT-qPCR. We found that LPS declined cell viability and heightened apoptosis and levels of Bax, cleaved-caspase-3, IL-6, and MCP-1. MIAT2 was negatively regulated by LPS and it alleviated LPS-induced damage. Furthermore, MIAT2 reversely regulated miR-15 and miR-15 mimic could reverse the effects of MIAT2. Finally, MIAT2 restrained the p38MAPK and NF-κB pathways by downregulating miR-15. In conclusion, MIAT2 alleviated LPS-induced inflammation damage in WI-38 cells by sponging miR-15 via p38MAPK and NF-κB pathways.
Assuntos
Inflamação/genética , MicroRNAs/genética , Pneumonia/genética , RNA Longo não Codificante/genética , Apoptose , Caspase 3/genética , Linhagem Celular , Sobrevivência Celular/genética , Quimiocina CCL2 , Regulação da Expressão Gênica/genética , Humanos , Recém-Nascido , Doenças do Recém-Nascido/genética , Doenças do Recém-Nascido/patologia , Inflamação/induzido quimicamente , Inflamação/patologia , Interleucina-6/genética , Lipopolissacarídeos/toxicidade , Pneumonia/patologia , Transdução de Sinais/genética , Fator de Transcrição RelA/genética , Proteína X Associada a bcl-2/genéticaRESUMO
BACKGROUND: Compared with Severe Acute Respiratory Syndrome (SARS) and Middle East Respiratory Syndrome (MERS), Corona Virus Disease 2019(COVID-19) spread more rapidly and widely. The population was generally susceptible. However, reports on pregnant women infected with SARS-CoV-2 were very limited. By sharing the clinical characteristics, treatments and outcomes of 18 patients with COVID-19 during late pregnancy, we hope to provide some references for obstetric treatment and management. METHODS: A total of 18 patients with COVID-19 treated at Renmin Hospital of Wuhan University were collected. The epidemiological characteristics, clinical manifestations, laboratory tests, chest CT and pregnancy outcomes were performed for analysis. RESULTS: 1. 18 cases of late pregnancy infected with SARS-CoV-2 pneumonia were delivered at 35 + 5 weeks to 41 weeks. According to the clinical classification of COVID-19, 1 case was mild type, 16 cases were ordinary type, and 1 case was severe type. 2. According to imaging examinations: 15 (83%) cases showed unilateral or bilateral pneumonia, 2 (11%) cases had pulmonary infection with pleural effusion, and 1 (6%) case had no abnormal imaging changes. 8 (44%) cases were positive and 10 (56%) cases were negative for nasopharyngeal-swab tests of SARS-CoV-2. 3. Among the 18 newborns, there were 3 (17%) premature infants, 1 (6%) case of mild asphyxia, 5 (28%) cases of bacterial pneumonia, 1 (6%) case of gastrointestinal bleeding, 1 (6%) case of necrotizing enteritis, 2 (11%) cases of hyperbilirubinemia and 1 (6%) case of diarrhea. All the newborns were negative for the first throat swab test of SARS-CoV-2 after birth. 4. Follow-up to Mar 7, 2020, no maternal and neonatal deaths occurred. CONCLUSIONS: The majority of patients in late term pregnancy with COVID-19 were of ordinary type, and they were less likely to develop into critical pneumonia after early isolation and antiviral treatment. Vertical transmission of SARS-CoV-2 was not detected, but the proportion of neonatal bacterial pneumonia was higher than other neonatal diseases in newborns.
Assuntos
Betacoronavirus , Infecções por Coronavirus/complicações , Transmissão Vertical de Doenças Infecciosas/estatística & dados numéricos , Pneumonia Viral/complicações , Complicações Infecciosas na Gravidez/virologia , Adulto , COVID-19 , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/virologia , Feminino , Humanos , Recém-Nascido , Pandemias , Pneumonia Viral/epidemiologia , Pneumonia Viral/virologia , Gravidez , Complicações Infecciosas na Gravidez/epidemiologia , Resultado da Gravidez , Nascimento Prematuro/epidemiologia , Nascimento Prematuro/virologia , SARS-CoV-2RESUMO
Neonatal pneumonia is mostly bacterial and other etiology is considered less frequently. We report a case of newborn whose neonatal pneumonia has not improved, despite the aggressive ventilation regime and empiric antibiotic therapy. A special sample from the respiratory tract was collected for PCR examination. The test confirmed the presence of Trichomonas vaginalis. Antibiotic therapy was extended to include metronidazole. Targeted antibiotic therapy, which lasted for 28 days, improved the condition and the patient was discharged in a stabilized condition to home care on the 44th day of life. We demonstrate the need to consider atypical pathogens in the case of infections that do not respond to conventional therapy. The multiplex real-time PCR technique was used to detect the DNA of the pathogen. Targeted antibiotic therapy is the result of pathogen identification.
Assuntos
Pneumonia , Tricomoníase , Trichomonas vaginalis , Humanos , Recém-Nascido , Metronidazol/uso terapêutico , Reação em Cadeia da Polimerase Multiplex , Tricomoníase/diagnóstico , Tricomoníase/tratamento farmacológico , Trichomonas vaginalis/genéticaRESUMO
Notoginsenoside R1 (NGR1) is a neoteric phytoestrogen extracted from Panax notoginseng, and possesses comprehensive pharmacological functions in multitudinous ailments. But, whether NGR1 is utilized in neonatal pneumonia is not clear. This research study aspired to disclose the protective activity of NGR1 in neonatal pneumonia. WI-38 cells were co-stimulated with NGR1 and lipopolysaccharide (LPS, 10 ng/mL), CCK-8 and flow cytometry assays were implemented for cell viability and apoptosis assessment. Real-time quantitative plymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA), and Western blot analysis were executed for inflammatory cytokine determination. MicroRNA-181a (miR-181a) expression was evaluated through RT-qPCR, simultaneously, the impact of miR-181a was estimated in NGR1 and LPS co-managed cells. Dual luciferase report assay was performed to disclose the relation between miR-181a and Toll-like receptor 4 (TLR4). The nuclear factor-κB (NF-κB) and TAK1/JNK pathways were ultimately appraised. We found that NGR1 decreased cell viability, evoked apoptosis and impeded interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α) expression and secretions in LPS-managed WI-38 cells. MiR-181a expression was enhanced by NGR1, and miR-181a inhibition inverted the impacts of NGR1 in LPS-managed WI-38 cells. Besides, TLR4 was predicted to be a firsthand direct target of miR-181a. Furthermore, NGR1 hindered NF-κB and TAK1/JNK pathways through modulating TLR4. These discoveries disclosed the fact that NGR1 protected WI-38 cells against LPS-triggered injury via adjusting the miR-181a/TLR4 and NF-κB and TAK1/JNK pathways.
Assuntos
Ginsenosídeos/farmacologia , Pulmão/citologia , MicroRNAs/metabolismo , Receptor 4 Toll-Like/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Lipopolissacarídeos/toxicidade , MAP Quinase Quinase Quinases/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NF-kappa B/metabolismo , Pneumonia/metabolismo , Pneumonia/patologia , Substâncias Protetoras/farmacologia , Receptor 4 Toll-Like/genéticaRESUMO
BACKGROUND: Phentolamine is an α-adrenergic receptor blocker that can be used to treat neonatal pneumonia, but its underlying mechanism is unclear. The purpose of this study is to probe the function of phentolamine on lipopolysaccharide (LPS)-induced inflammation and cell death in an in vitro model of neonatal pneumonia. METHODS: Human MRC-5 cells were incubated with varying doses of phentolamine in vitro to evaluate cell viability. Subsequently, LPS was introduced to further investigate the combined effects of phentolamine and LPS on cell viability and apoptosis in MRC-5 cells. The effect of phentolamine/LPS treatment on the Neurotrophic Tyrosine Kinase Receptor A (TrkA)/Protein Kinase B (Akt) signaling pathway and the phosphorylation of pathway proteins in MRC-5 cells was further analyzed via western blot. Additionally, knockout of TrkA and Akt genes in MRC-5 cells was performed to explore the effects of phentolamine/LPS treatment on cell viability, apoptosis levels, and inflammatory factor levels in MRC-5 cells. RESULTS: Preincubation of MRC-5 cells with a low concentration of phentolamine (≤6 µg/mL) protected against LPS-induced cell inflammatory injury. Phentolamine promoted both TrkA and Akt phosphorylation and Akt activation induced by LPS in MRC-5 cells. The protective effect of phentolamine against LPS-induced apoptosis and inflammation was significantly reduced in response to TrkA or Akt gene knockdown in MRC-5 cells. CONCLUSIONS: Phentolamine may protect LPS-induced apoptosis and inflammation by activating the TrkA and Akt signaling pathways.
Assuntos
Lipopolissacarídeos , Pneumonia , Recém-Nascido , Humanos , Lipopolissacarídeos/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/farmacologia , Fentolamina/farmacologia , Transdução de Sinais , Inflamação/metabolismo , ApoptoseRESUMO
BACKGROUND: Excessive inflammation is the main cause of treatment failure in neonatal pneumonia (NP). CCCTC-binding factor (CTCF) represents an important node in various inflammatory diseases. In the present study, we tried to clarify the function and underlying molecular mechanism of CTCF on an in vitro cellular model of NP, which was generated by simulating the human lung fibroblast cell line WI-38 with lipopolysaccharide (LPS). METHODS: The SUMOylation level and protein interaction were verified by Co-immunoprecipitation assay. Cell viability was measured by Cell Counting Kit-8 assay. Inflammatory factors were examined by Enzyme-linked immunosorbent assay. Cell apoptosis was evaluated by TUNEL assay. The binding activity of CTCF to target promoter was tested by chromatin immunoprecipitation and luciferase reporter assay. RESULTS: LPS treatment restrained cell viability, promoted the production of inflammatory factors, and enhanced cell apoptosis. CTCF overexpression played anti-inflammatory and anti-apoptotic roles. Furthermore, CTCF was modified by SUMOylation with small ubiquitin-like modifier protein 1 (SUMO1). Interfering with sumo-specific protease 1 (SENP1) facilitated CTCF SUMOylation and protein stability, thus suppressing LPS-evoked inflammatory and apoptotic injuries. Moreover, CTCF could bind to the forkhead box protein A2 (FOXA2) promoter region to promote FOXA2 expression. The anti-inflammatory and anti-apoptotic roles of CTCF are associated with FOXA2 activation. In addition, SENP1 knockdown increased FOXA2 expression by enhancing the abundance and binding ability of CTCF. CONCLUSIONS: SUMOylation of CTCF by SENP1 knockdown enhanced its protein stability and binding ability and it further alleviated LPS-evoked inflammatory injury in human lung fibroblasts by positively regulating FOXA2 transcription.
Assuntos
Lipopolissacarídeos , Peptídeo Hidrolases , Recém-Nascido , Humanos , Peptídeo Hidrolases/metabolismo , Fator de Ligação a CCCTC/genética , Fator de Ligação a CCCTC/metabolismo , Sumoilação , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Fibroblastos/metabolismo , Pulmão/metabolismo , Anti-Inflamatórios , Fator 3-beta Nuclear de Hepatócito/genética , Fator 3-beta Nuclear de Hepatócito/metabolismoRESUMO
AIM: Pulmonary near-infrared spectroscopy (NIRS) is a new and promising tool for diagnosis of neonatal respiratory diseases (RD). The study aimed to determine the role of pulmonary regional oxygen saturation (pRSO2 ) values obtained by NIRS in the early distinction of neonatal pneumonia (NP) from transient tachypnea of the newborn (TTN). METHODS: This prospective, observational, double-blind study was conducted in neonatal intensive care unit (NICU) between 2020 and 2021. Late preterm and term newborns hospitalized in the NICU due to the diagnosis of TTN and NP were included. Cerebral RSO2 and pRSO2 values were measured during the 1st, 24th, 48th, and 72nd hours of hospitalization, using NIRS. RESULTS: Of the eligible 40 infants, 65% (n: 26) were diagnosed as TTN and 35% (n: 16) as NP. The pRSO2 values were significantly higher in the TTN group than the NP group for both apexes (75.3 ± 8.7 vs. 69 ± 5.4, p: .018, respectively) and lateral lung (77.8 ± 6 vs. 72.7 ± 6.2, p: .016, respectively) in the 1st hour of hospitalization. There were significant differences in pRSO2 apex and pRSO2 lateral values between the 1st and 24th hours of hospitalization and the 24th and 48th hours in the NP group (p2 : .001 for both). The optimal pRSO2 apex cut-off value was >72% to predict the diagnosis of NP with a sensitivity of 78.6% and a specificity of 69.2%. CONCLUSION: Pulmonary NIRS may be considered as a feasible and promising diagnostic tool in late preterm and term infants with RD. It may also be helpful for the early differentiation of NP from TTN and the courses of these diseases.
Assuntos
Pneumonia , Taquipneia Transitória do Recém-Nascido , Humanos , Recém-Nascido , Pulmão/diagnóstico por imagem , Pneumonia/diagnóstico , Estudos Prospectivos , Espectroscopia de Luz Próxima ao Infravermelho , Taquipneia Transitória do Recém-Nascido/diagnóstico , Método Duplo-CegoRESUMO
Human respiratory syncytial virus (HRSV) is a major pathogen of lower respiratory tract infections in children (<5 years) and older individuals, with outbreaks mainly reported among infants in hospital pediatric departments and intensive care units (ICUs). An outbreak of severe neonatal pneumonia occurred in a postpartum center in Shenyang city, China, from January to February 2021. In total, 34 respiratory samples were collected from 21 neonates and 13 nursing staff. The samples were screened for 27 pathogens using a TaqMan low-density array, and 20 samples tested positive for HRSV, including 16 neonates and 4 nursing staff samples. Among the 16 hospitalized neonates, seven were admitted to an ICU and nine to general wards. Four of the nursing staff had asymptomatic infections. To investigate the genetic characteristics of the HRSV responsible for this outbreak, the second hypervariable region (HVR2) sequences of the G gene were obtained from six neonates and two nursing staff. Phylogenetic analyses revealed that all eight sequences (SY strains) were identical, belonging to the HRSV BA9 genotype. Our findings highlight the necessity for strict hygiene and disease control measures so as to prevent cross-infection and further avoid potential outbreaks of severe infectious respiratory diseases. IMPORTANCE Human respiratory syncytial virus (HRSV) is one of the leading causes of acute lower respiratory infections (ALRI) worldwide. In this study, we first reported an outbreak of severe neonatal pneumonia caused by HRSVB BA9 at a postpartum care center in mainland China. Among 20 confirmed cases, 16 were hospitalized neonates with 7 in the ICU ward, and the other four were nursing staff with asymptomatic infections. Our findings highlighted the importance of preventing cross-infection in such postpartum centers.
Assuntos
Pneumonia , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Infecções Respiratórias , Infecções Assintomáticas/epidemiologia , Criança , China/epidemiologia , Surtos de Doenças , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Filogenia , Pneumonia/epidemiologia , Cuidado Pós-Natal , Gravidez , Infecções por Vírus Respiratório Sincicial/epidemiologia , Vírus Sincicial Respiratório Humano/genética , Infecções Respiratórias/epidemiologiaRESUMO
OBJECTIVE: In this work, we investigated the effects of gambogic acid (GA) on lipopolysaccharide (LPS)-induced apoptosis and inflammation in a cell model of neonatal pneumonia. METHOD: Human WI-38 cells were maintained in vitro and incubated with various concentrations of GA to examine WI-38 survival. GA-preincubated WI-38 cells were then treated with LPS to investigate the protective effects of GA on LPS-induced death, apoptosis and inflammation. Western blot assay was utilized to analyze the effect of GA on tropomyosin receptor kinase A (TrkA) signaling pathway in LPS-treated WI-38 cells. In addition, human AKT serine/threonine kinase 1 (Akt) gene was knocked down in WI-38 cells to further investigate the associated genetic mechanisms of GA in protecting LPS-induced inflammation and apoptosis. RESULTS: Pre-incubating WI-38 cells with low and medium concentrations GA protected LPS-induced cell death, apoptosis and inflammatory protein productions of IL-6 and MCP-1. Using western blot assay, it was demonstrated that GA promoted TrkA phosphorylation and Akt activation in LPS-treated WI-38 cells. Knocking down Akt gene in WI-38 cells showed that GA-associated protections against LPS-induced apoptosis and inflammation were significantly reduced. CONCLUSIONS: GA protected LPS-induced apoptosis and inflammation, possibly through the activations of TrkA and Akt signaling pathway. This work may broaden our understanding on the molecular mechanisms of human neonatal pneumonia.
Assuntos
Anti-Inflamatórios/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor trkA/metabolismo , Xantonas/farmacologia , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular , Humanos , Recém-Nascido , Lipopolissacarídeos , Pneumonia , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Emerging evidence suggests that long non coding RNA (lncRNA) small nucleolar RNA host gene 4 (SNHG4) has become a new insight into lipopolysaccharide (LPS)-induced microglia inflammation, its role in neonatal pneumonia (NP) remains to be largely unrevealed. METHODS: RT-qPCR was used to determine the expression of SNHG4 and METTL3 in the serum from NP patients and normal volunteers, as well as in LPS treated-WI-38 cells. The SNHG4 overexpression vector (pcDNA-SNHG4) was transfected into LPS-treated cells. CCK-8, Transwell, annexin V-FITC/PI, ELISA and Western blot assays were used to determine cell proliferation, migration, apoptosis, contents of IL-6, TNF-α, SOD and MDA, as well as the expression levels of NF-κB pathway proteins, respectively. The enrichment of SNHG4 in the METTL3 promoter region was assessed with RIP assay. m6A quantitative analysis illustrated the m6A level with or without SNHG4 overexpression or METTL3 silencing. Bioinformatics analysis and RIP-PCR were used to predict and validate YTHDF1-mediated m6A levels on signal transducer and activator of transcription 2 (STAT2) mRNA in METTL3 inhibited cells. Then rescue experiments were performed to explore effects of SNHG4 and METTL3 or STAT2 on LPS-treated cell functions. Subsequently, in vivo functional experiments were performed to investigate the role of SNHG4 in LPS induced pneumonia in mice. RESULTS: SNHG4 was downregulated, and METTL3 was upregulated in NP patients and LPS-treated cells. SNHG4 overexpression facilitated cell proliferation, migration and SOD concentration, as well as inhibited cell apoptosis and production of IL-6, TNF-α and MDA, and suppressed the expression of NF-κB pathway proteins. Mechanistically, SNHG4 bound with METTL3 and downregulated METTL3 expression. Besides, total m6A level was lower in the SNHG4 overexpressed or METTL3 inhibited cells. METTL3 interference reduced m6A levels of STAT2 mRNA, decreased STAT2 mRNA stability and promoted STAT2 translation level. Upregulation of METTL3 or STAT2 reversed the effects of SNHG4 overexpression on LPS-treated cell functions. CONCLUSIONS: This study reveals that SNHG4 promotes LPS induced inflammation in human lung fibroblasts and mouse lung tissues in vitro and in vivo by inhibiting METTL3-mediated m6A level of STAT2 mRNA, which may provide a potential therapeutic mechanism for NP.
Assuntos
Inflamação/imunologia , Metiltransferases/metabolismo , Pneumonia/imunologia , RNA Longo não Codificante/metabolismo , Animais , Feminino , Regulação da Expressão Gênica/imunologia , Humanos , Recém-Nascido , Doenças do Recém-Nascido/imunologia , Doenças do Recém-Nascido/patologia , Inflamação/patologia , Lipopolissacarídeos/imunologia , Masculino , Camundongos , Pneumonia/patologia , RNA Mensageiro/metabolismo , Fator de Transcrição STAT2/metabolismoRESUMO
BACKGROUND: Possible impact of maternal hypertension and preeclampsia on neonatal respiratory disorders was unknown. We investigated the association of maternal hypertension and preeclampsia with neonatal respiratory disorders in preterm and full-term newborns. METHOD: In this study, we used a large Chinese population-based study which includes 185,687 singleton livebirths with gestational weeks between 28 and 42 weeks. The "exposure" was maternal hypertension and preeclampsia. The outcome was neonatal respiratory disorders including neonatal respiratory distress syndrome (NRDS), pneumonia and low Apgar scores. Logistic regression was used to examine the association between the maternal hypertensive disorders and the risk for neonatal respiratory disorders. We further evaluated the association in preterm and full-term infants. RESULTS: As a result, the incidence of neonatal RDS, pneumonia, and low Apgar score in hypertensive group was higher than that in normotensive group. Preeclampsia was associated with RDS [(adjusted relative risk (aRR): 1.85, 95% confidence interval (CI): 1.22-3.05)]. Both maternal hypertension and preeclampsia increased risks for neonatal pneumonia (aRR: 1.79, 95%CI: 1.48-2.17; aRR: 1.81, 95%CI: 1.36-2.40, respectively), for low Apgar score at 1 min (aRR: 1.20, 95%CI: 1.13-1.27; aRR: 1.53, 95%CI: 1.41-1.67, respectively), and for low Apgar score at 5 min (aRR: 1.30, 95%CI: 1.17-1.45; aRR: 1.70, 95%CI: 1.46-1.99, respectively). The risk for neonatal respiratory disorders increased with severity of maternal hypertension. The observed associations were present in both full-term and preterm birth. CONCLUSION: Maternal hypertension and preeclampsia are risk factors for neonatal respiratory disorders in full-term and preterm newborns.
Assuntos
Hipertensão/epidemiologia , Pneumonia/epidemiologia , Pré-Eclâmpsia/epidemiologia , Complicações Cardiovasculares na Gravidez/epidemiologia , Síndrome do Desconforto Respiratório do Recém-Nascido/epidemiologia , Índice de Apgar , China/epidemiologia , Estudos de Coortes , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Gravidez , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
BACKGROUND: Neonatal pneumonia (NP) has a high fatality rate in neonatal illness. This research investigated the functions of emodin on lipopolysaccharide (LPS)-evoked inflammatory injury in WI-38 cells. METHODS: Cell counting kit-8 (CCK-8) assay and flow cytometry were utilized for examining the impacts of LPS and emodin on viability and apoptosis, respectively. Taurine up-regulated gene 1 (TUG1) level was altered through cell transfection and investigated by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Moreover, RT-qPCR, western blot and enzyme-linked immunosorbent assay (ELISA) were utilized for investigating expressions of monocyte chemoattractant protein-1 (MCP-1) and interleukin (IL)-6. Western blot was carried out for investigating the levels of Bcl-2, Bax, pro-Caspase-3, cleaved-Caspase-3 and NF-κB and p38MAPK pathway-related proteins. RESULTS: LPS treatment restrained cell viability, enhanced apoptosis, and expressions of inflammation-related IL-6 and MCP-1. Emodin alleviated LPS-evoked inflammatory injury and restrained the NF-κB and p38MAPK pathways. Furthermore, emodin positively regulated TUG1 expression and TUG1 silencing could reverse the efficacy of emodin on IL-6 and MCP-1 expressions. Finally, TUG1 regulates the expression of inflammatory factors through NF-κB and p38MAPK pathways. CONCLUSION: Emodin alleviated LPS-evoked inflammatory injury by raising TUG1 expression via NF-κB and p38MAPK pathways in WI-38 cells.
Assuntos
Emodina/farmacologia , Inflamação/tratamento farmacológico , RNA Longo não Codificante/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quimiocina CCL2/genética , Humanos , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/patologia , Interleucina-6/genética , Lipopolissacarídeos/toxicidade , NF-kappa B/genética , RNA Longo não Codificante/antagonistas & inibidores , Taurina/genética , Ativação Transcricional/efeitos dos fármacosRESUMO
BACKGROUND: Neonatal pneumonia is a common disease in the neonatal period with a high incidence and death. This study aimed to investigate the molecular mechanism and effect of microRNA (miR)-429 in neonatal pneumonia. METHODS: The peripheral blood was collected from neonatal pneumonia and healthy patients, respectively. Human lung fibroblast WI-38 cells were treated with lipopolysaccharide (LPS) to establish neonatal pneumonia cell model. Then, the miR-429 expression was detected by quantitative real-time polymerase chain reaction (qRT-PCR). In addition, the relationship between miR- 429 and kruppel-like factor 4 (KLF4) was confirmed by dual luciferase reporter assay. Cell viability, the level of interleukin 6 (IL-6), IL-1ß and tumor necrosis factor α (TNF-α) and apoptosis were measured by Cell Counting Kit-8 (CCK-8), enzyme linked immunosorbent assay (ELISA) and flow cytometry. Meanwhile, apoptosis and nuclear factor kappa-B (NF-κB) pathway related proteins expression were analyzed by western blot. RESULTS: MiR-429 expression level was increased in neonatal peripheral blood and LPS-stimulated WI-38 cells. Then, miR-429 overexpression increased apoptosis, the level of IL-6, IL-1ß, TNF-α, Bax and cleaved caspase-3, while reduced cell viability in LPS-stimulated WI-38 cells. Besides, KLF4 was identified as the target gene of miR-429, and reversed the changes caused by miR-429 overexpression. Finally, miR-429 suppressor down-regulated p-NF-κB level in LPS-stimulated cells and KLF4 knockdown reversed these reductions. CONCLUSION: MiR-429 promotes inflammatory injury, apoptosis and activates the NF-κB signaling pathway by targeting KLF4 in neonatal pneumonia, and then these results provide evidence for clinical diagnosis and treatment for neonatal pneumonia.
Assuntos
Fatores de Transcrição Kruppel-Like/metabolismo , Pulmão/metabolismo , MicroRNAs/metabolismo , Pneumonia/sangue , Apoptose/fisiologia , Sobrevivência Celular/fisiologia , Feminino , Humanos , Recém-Nascido , Inflamação , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Fator 4 Semelhante a Kruppel , Lipopolissacarídeos/farmacologia , Pulmão/citologia , Pulmão/efeitos dos fármacos , Masculino , NF-kappa B/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
AIMS: The aims of this study were to explore factors that associated with gestational diabetes mellitus (GDM), and to determine the relationship between early maternal HbA1c levels and adverse fetal or neonatal events, and to determine an optimal maternal glucose testing method in order to decrease the potential health risk for their offspring. METHODS: From December 2015 to May 2016, a total of 6744 pregnant women were enrolled from Shanghai First Maternal and Infant Hospital affiliated to Tongji University prospectively in the nested case-control study. Each GDM case was matched with a healthy pregnant woman and followed up. Outcome analyses were conducted between GDM case and control groups, as well as elevated and normal maternal HbA1c levels, respectively. RESULTS: A total of 1836 women were included in the adverse fetal and neonatal events examination. For pregnant women with early HbA1câ¯≥â¯5.2%, the adjusted risk ratios (RR) of respiratory distress syndrome (RDS), pneumonia and jaundice were 4.37 (95%CI 1.54-12.35), 2.03 (95%CI 1.24-3.33) and 1.49 (95%CI 1.01-2.20), respectively. After treatments, the frequency for the majority of events in GDM group was similar to that of healthy pregnant women. Moreover, the area under the curve (AUC) of early maternal HbA1c in predicting potential RDS is 0.734. HbA1câ¯≤â¯4.9% excluded for RDS. CONCLUSIONS: Compared with women with normal HbA1c, those with an early elevated HbA1c level were more likely to develop adverse events, including RDS, pneumonia and jaundice. Early HbA1c testing can be used as an auxiliary method identifying potential RDS.
Assuntos
Diabetes Gestacional/sangue , Doenças Fetais/etiologia , Hemoglobinas Glicadas/metabolismo , Síndrome do Desconforto Respiratório do Recém-Nascido/etiologia , Adulto , Estudos de Casos e Controles , Estudos de Coortes , Feminino , Doenças Fetais/sangue , Humanos , Recém-Nascido , GravidezRESUMO
Psittacosis is a rare zoonosis that can cause severe disease and adverse outcomes during pregnancy. We identified a previously elusive case of psittacosis causing premature delivery and infant death by next-generation RNA sequencing of postmortem tissues. Hypothesis-free pathogen detection in postmortem specimens can increase the yield of epidemiologic and cause-of-death studies.
RESUMO
Mycoplasma pneumoniae is a significant cause of pneumonia in school-aged children and young adults. We report a case of neonatal M. pneumoniae pneumonia in a preterm child manifesting in the first hours of life. Vertical transmission was demonstrated by the detection of M. pneumoniae in inflamed placental tissue indicating chorioamnionitis.