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BACKGROUND: Midwives experience elevated levels of work-related stress. However, there is limited understanding of the patterns of this stress, its sociodemographic and work-related determinants, and its association with sub-health. METHOD: This multi-center cross-sectional study, conducted in 21 hospitals in Zhuhai City, Guangdong Province, China, involved 300 midwives. Work-related stress levels in midwives were evaluated using the Chinese version of the Nursing Stress Scale, while social, physical and mental sub-health status was measured with the Sub-Health Measurement Scale. Utilizing latent profile analysis, the study aimed to categorize midwives into homogeneous profiles based on patterns of work-related stress. RESULTS: Among the 300 midwives examined, three distinct profiles were identified: profile 1 (n=57, 19â¯%), characterized by low work-related stress; profile 2 (n=149, 50â¯%), representing the moderate work-related stress class; and profile 3 (n=94, 31â¯%), indicative of high work-related stress. Midwives in the high work-related stress profile tended to be younger, with lower monthly income, lower professional titles, and a higher likelihood of night shift work (all P<0.01). Significant and noteworthy trends were observed in sociodemographic characteristics (age, monthly income, and professional title) and work-related characteristics (night shift status). After controlling for confounders, the work-related stress profile demonstrated a negative association with social, physical and mental sub-health status. CONCLUSION: This study highlights integrating stress domains and adopting a person-centered approach to examine midwives' work-related stress. Identifying predictors of profile membership and their relationship with sub-health can inform tailored interventions to reduce stress and improve midwives' well-being.
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ETHNOPHARMACOLOGICAL RELEVANCE: The Qinglongyi-Buguzhi herbal pair (QB) is one of commonly used herbal combinations for treating vitiligo in traditional Chinese medicine, consisting of the exocarp of the immature fruit of Juglans regia L. or Juglans mandshurica Maxim., and dried, mature fruit of Psoralea corylifolia L. However, the active components and potential mechanisms of QB in the treatment of vitiligo are still unclear. AIM OF THE STUDY: The purpose of this study is to clarify the effects and mechanisms of QB on vitiligo treatment through integration of network pharmacology and empirical examinations. MATERIALS AND METHODS: The active components and targets of QB as well as the targets linked to vitiligo were obtained from network databases. Visualization networks were constructed with Cytoscape 3.9.1. GO and KEGG enrichment analysis were conducted to investigate the possible mechanism. Molecular docking was employed to evaluate the binding affinities between the primary active ingredients of QB and essential targets of the PI3K/Akt/Nrf2 pathway. In vivo and in vitro experiments were carried out to confirm the results of network pharmacology. RESULTS: We evaluated 44 active compounds and 602 genes from QB, and 107 of these genes linked to vitiligo. GO analysis suggested QB might lessen vitiligo by regulating oxidative stress. KEGG pathway analysis indicated the PI3K/Akt pathway may be crucial for treating vitiligo. Molecular docking results demonstrated the key active ingredients of QB had good binding activity with the major targets in the PI3K/Akt/Nrf2 pathway. In vivo, QB significantly ameliorated vitiligo model mouse's skin pathologies by reducing ROS, elevating CAT and SOD levels. Western blot showed that QB increased the phosphorylation of PI3K and Akt and the expressions of Nrf2 and HO-1 in the skin. In vitro, QB reversed H2O2-induced oxidative injury of melanocytes, enhanced cell survival rate, reduced ROS level, upregulated SOD and CAT activities, and raised the content of melanin. Moreover, QB upregulated the expression levels of Akt, Nrf2, HO-1 mRNA, Akt phosphorylation, HO-1, and nuclear Nrf2 proteins, and also encouraged the nuclear translocation of Nrf2. However, LY294002 treatment significantly reversed the regulatory effect of QB on oxidative damage of melanocytes. CONCLUSIONS: This study revealed that the therapeutic effect of QB on vitiligo was achieved through multiple components, targets and pathways. Experimental investigation demonstrated that QB could improve vitiligo via reducing oxidative stress, which was probably accomplished by activating the PI3K/Akt/Nrf2 signaling pathway.
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Medicamentos de Ervas Chinesas , Simulação de Acoplamento Molecular , Fator 2 Relacionado a NF-E2 , Farmacologia em Rede , Vitiligo , Vitiligo/tratamento farmacológico , Animais , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/uso terapêutico , Fator 2 Relacionado a NF-E2/metabolismo , Camundongos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Masculino , Fosfatidilinositol 3-Quinases/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , HumanosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Sophora flavescens Ait.-Angelica sinensis(Oliv.) Diels drug pairing (SA) is a transformed drug pairing from Shengui pill, a traditional Chinese medicine prescription in the ninth volume of Traditional Chinese Medicine classic "Gu Jin Yi Jian", which is famous for clearing heat, moistening dryness, and promoting blood circulation. It is commonly used in the treatment of eczema, a skin condition that causes itching and inflammation. Despite its widespread use, there is still limited research on the mechanism of how SA treats eczema. This paper aims to fill this gap by conducting animal experiments to uncover the mechanism behind SA's therapeutic effects on eczema. Our findings provide a solid foundation for the clinical use of this TCM prescription. AIM OF THE STUDY: The basic purpose of this study is to clarify the therapeutic mechanism of Sophora flavescens-Angelica sinensis (SA) in the treatment and control of eczema. MATERIALS AND METHODS: The chemical compositions of SA were analyzed using HPLC-Q-Orbitrap-MS. In vivo, a mouse model of eczema was created, and the serum levels of TNF-α and IL-1ß were quantified using an enzyme-linked immunosorbent assay (ELISA). Hematoxylin and eosin (HE) staining was performed to assess the pathological state of the mouse skin, and immunohistochemical technique (IHC) was employed to estimate the contents of TNF-α, TLR4, and NF-κB semi-quantitatively. The expression levels of TLR4, MyD88, and NF-κB mRNA were determined through real-time quantitative polymerase chain reaction (qRT-PCR). Western Blotting was utilized to identify the protein levels of TLR4, MyD88, and NF-κB in mouse skin tissue. RESULTS: SA identified 18 active chemicals, some of which were shown in vivo to inhibit the TLR4/MyD88/NF-κB signaling pathway while reducing serum levels of TNF-α and IL-1ß, making them ideal agents for the treatment of eczema. CONCLUSIONS: SA's anti-inflammatory properties are attributed to its ability to reduce serum levels of TNF-α and IL-1ß, likewise inhibit the TLR4/MyD88/NF-κB signaling pathway.
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Angelica sinensis , Eczema , Camundongos , Animais , NF-kappa B/metabolismo , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Sophora flavescens , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Primary cutaneous CD4+ small and medium pleomorphic T-cell lymphoproliferative disorder (PCSM-LPD) is a rare and typically asymptomatic proliferation of CD3+/CD4+ small and medium pleomorphic T-cells. In this case report, we share the details of a 41-year-old male presenting with a two-centimeter soft, mobile forehead nodule that was determined by clinical symptoms, histology, and immunostaining to be PCSM-LPD. We would like to emphasize the clinical resolution that was seen with minimal treatment.
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Systemic anaplastic lymphoma kinase-negative (ALK-) anaplastic large cell lymphoma (ALCL) comprises a genomically heterogeneous disease that is considered a distinct entity by the 2016 World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues. Other than lymph nodes, systemic ALK- ALCL may affect extranodal tissues, sites where the inflammatory background may be especially prominent. In this scenario, myxoid change is exceptional in systemic ALK- ALCL. We describe a rare case of systemic ALK- ALCL with distinctive myxoid changes, carrying specific chromosomal aberrations that affect the clinical outcome. Careful morphological, immunohistochemical, and molecular workup is mandatory because a myxoid background should not be a reason to ignore the possibility of a lymphoma. Finally, extensive correlation with staging and the detection of prognostic biomarkers such as DUSP22 and TP63 rearrangements are essential for the diagnosis and prediction of clinical outcome in ALK- ALCL.
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Linfoma Anaplásico de Células Grandes , Humanos , Linfoma Anaplásico de Células Grandes/patologia , Rearranjo Gênico , Linfonodos/patologia , Fosfatases de Especificidade Dupla/genética , Fosfatases da Proteína Quinase Ativada por Mitógeno/genéticaRESUMO
Interleukin (IL)-33 is a tissue-derived nuclear cytokine belonging to the IL-1 family. Stimulation-2 (ST2) is the only known IL-33 receptor. ST2 signals mostly on immune cells found within tissues, such as regulatory T cells (Treg cells), CD8+ T cells, and natural killer (NK) cells. Therefore, the IL-33/ST2 signaling pathway is important in the immune system. IL-33 deficiency impairs Treg cell function. ST2 signaling is also increased in active Treg cells, providing a new approach for Treg-related immunotherapy. The IL-33/ST2 signaling pathway regulates multiple immune-related cells by activating various intracellular kinases and factors in the tumor microenvironment (TME). Here, we review the latest studies on the role of the IL-33/ST2 signaling pathway in TME and Treg immunotherapy.
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Proteína 1 Semelhante a Receptor de Interleucina-1 , Linfócitos T Reguladores , Microambiente Tumoral , Interleucina-33 , ImunoterapiaRESUMO
The incidence rate of breast cancer is the highest in the world, and major problem in the clinical treatment is the therapy resistance of breast cancer stem cells (CSCs). Thus, new therapeutic approaches targeting breast CSCs are needed. Our previous study demonstrated cancer-derived sialylated IgG (SIA-IgG) is highly expressed in cancer cells with stem/progenitor features. Furthermore, a high frequency of SIA-IgG in breast cancer tissue predicted metastasis and correlated with poor prognosis factors, and depletion of IgG in breast cancer leads to lower malignancy of cancer cells, suggesting SIA-IgG could be a potential therapeutic target in breast cancer. In this study, we first investigated the relationship of SIA-IgG expression with the clinicopathological characteristics and clinical prognosis of breast carcinoma patients, and the data confirmed that the expression of SIA-IgG confers poor prognosis in breast cancer. Successively, by using a monoclonal antibody specifically against SIA-IgG, we targeted SIA-IgG on the surface of MDA-MB-231 cells and detected their functional changes, and the results suggested SIA-IgG to be a promising antibody therapeutic target in breast cancer. In addition, we explored the mechanism of action at the molecular level of SIA-IgG on breast cancer cell, the findings suggest that SIA-IgG promotes proliferation, metastasis, and invasion of breast cancer cells through the Wnt/ß-catenin signaling pathway. Developing therapeutic antibody needs effective therapeutic target, and the antibody should better be a monoclonal antibody with high affinity and high specificity. This study provides a potential prognostic marker and a novel therapeutic target for breast cancer.
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Neoplasias da Mama , Anticorpos Monoclonais/uso terapêutico , Neoplasias da Mama/metabolismo , Feminino , Humanos , Imunoglobulina G , Prognóstico , Via de Sinalização WntRESUMO
As flipped classroom has received much attention from researchers and educators, some scholars have investigated the effectiveness of this teaching mode in various English as a foreign language (EFL) settings; however, such an instruction mode has been under-investigated in the Chinese EFL context. Therefore, the current study examined a flipped course's impact on Chinese EFL learners' writing performance and anxiety utilizing a pretest-posttest non-equivalent group quasi-experimental design. First, it selected a sample of 50 Chinese EFL learners from two intact language school classes as the participants via the convenience sampling method. Then, it randomly assigned participants of these two intact classes to a control group (n = 24), taught based on the traditional method of writing instruction, and an experimental group (n = 26), instructed based on social media-supported flipped instruction. The study used two writing tasks and a writing anxiety inventory to gather the data from the participants. The descriptive and inferential statistics results showed that the experimental group-taught based on flipped writing instruction-significantly enhanced their writing performance. Moreover, they revealed that the flipped course substantially reduced participants' writing anxiety. Implications of such findings have been elaborated for EFL research and practice.
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Tripterygium hypoglaucum hutch (THH) is a plant of the genus tripterygium, which is also known as colquhounia, Gelsemiun elegan, and so on. It is mainly distributed in Yunnan, Guizhou, and Sichuan regions and other places in China. To study the immune mechanism of THH on related inflammatory cytokines in collagen II-induced arthritis (CIA) mice, healthy male C57BL/6 mice were used to model CIA mice. Mice received THH 420 mg/kg/day or the same amount of normal saline (NS) by gavage for 20 days. The thickness of the ankle joint in mice was observed, and the arthritis index was calculated. Related inflammatory cytokines were detected by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. The results showed that after treatment with THH, the CIA mice had less swelling and destruction of the joints as well as decreased foot size and arthritis index. The mRNA and protein levels of TNF-α, IFN-γ, and IL-17A were lower in the THH-treated group than in the NS group (P < 0.05). In summary, THH has great significance in the treatment of CIA mice, including reduced related inflammatory cytokines expression level in both joint tissue and serum. The mechanism of THH in the treatment of CIA may be through the inhibition of the NF-kB-STAT3-IL-17 pathway, which also requires further experimental investigation.
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Artrite Experimental/metabolismo , Citocinas/metabolismo , Inflamação , Extratos Vegetais/farmacologia , Tripterygium/química , Animais , Peso Corporal , China , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Interferon gama/metabolismo , Interleucina-17/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Subunidade p50 de NF-kappa B/metabolismo , RNA Mensageiro/metabolismo , Fator de Transcrição STAT3/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Anaplastic lymphoma kinase negative systemic anaplastic large cell lymphoma (ALK-ALCL) is a CD30+ T-cell malignant lymphoma which may involve both lymph nodes and extranodal tissues, showing important clinical differences from ALK-positive ALCL (ALKâ¯+â¯ALCL). ALK- ALCL is considered a specific entity by the 2016 World Health Organization (WHO) classification of hematolymphoid neoplasms.We describe an exceptional case of ALK- ALCL with a striking "Hodgkin-like" cytomorphology and a very uncommon nuclear expression of PAX5.
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Linfoma Anaplásico de Células Grandes/diagnóstico , Fator de Transcrição PAX5/metabolismo , Idoso de 80 Anos ou mais , Núcleo Celular/metabolismo , Feminino , Humanos , Linfonodos/metabolismo , Linfonodos/patologia , Linfoma Anaplásico de Células Grandes/metabolismo , Linfoma Anaplásico de Células Grandes/patologia , Fator de Transcrição PAX5/genéticaRESUMO
Macrophages are divided into two subpopulations: classically activated macrophages (M1) and alternatively activated macrophages (M2). BCG (Bacilli Calmette-GuC)rin) activates disabled naC/ve macrophages to M1 macrophages, which act as inflammatory, microbicidal and tumoricidal cells through cell-cell contact and/or the release of soluble factors. Various transcription factors and signaling pathways are involved in the regulation of macrophage activation and polarization. We discovered that BCG-activated macrophages (BAM) expressed a new molecule, and we named it Novel Macrophage Activated Associated Protein 1 (NMAAP1). The current study found that the overexpression of NMAAP1 in macrophages results in M1 polarization with increased expression levels of M1 genes, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF-N1), Interleukin 6 (IL-6), Interleukin 12 (IL-12), Monocyte chemoattractant protein-1 (MCP-1) and Interleukin-1 beta (IL-1N2), and decreased expression of some M2 genes, such as Kruppel-like factor 4 (KLF4) and suppressor of cytokine signaling 1 (SOCS1), but not other M2 genes, including arginase-1 (Arg-1), Interleukin (IL-10), transforming growth factor beta (TGF-N2) and found in inflammatory zone 1 (Fizz1). Moreover, NMAAP1 overexpression in the RAW264.7 cell line increased cytotoxicity against MCA207 tumor cells, which depends on increased inflammatory cytokines rather than cell-cell contact. NMAAP1 also substantially enhanced the phagocytic ability of macrophages, which implies that NMAAP1 promoted macrophage adhesive and clearance activities. Our results indicate that NMAAP1 is an essential molecule that modulates macrophages phenotype and plays an important role in macrophage tumoricidal functions.
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Citotoxicidade Imunológica , Ativação de Macrófagos , Macrófagos/imunologia , Proteínas de Membrana/metabolismo , Mycobacterium bovis/imunologia , Neoplasias/imunologia , Animais , Linhagem Celular Tumoral , Quimiocina CCL2/metabolismo , Interleucinas/metabolismo , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos ICR , Óxido Nítrico Sintase Tipo II/metabolismo , Fagocitose , Células RAW 264.7 , Proteína 1 Supressora da Sinalização de Citocina , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Previous studies showed that TNF-α could activate voltage-gated Na(+) channels (VGSCs) in the peripheral nervous system (PNS). Since TNF-α is implicated in many central nervous system (CNS) diseases, we examined potential effects of TNF-α on VGSCs in the CNS. METHODS: Effects of TNF-α (1-1000 pg/mL, for 4-48 h) on VGSC currents were examined using whole-cell voltage clamp and current clamp techniques in primary culture of mouse cortical neurons. Expression of Nav1.1, Nav1.2, Nav1.3, and Nav1.6 were examined at both the mRNA and protein levels, prior to and after TNF-α exposure. RESULTS: TNF-α increased Na(+) currents by accelerating the activation of VGSCs. The threshold for action potential (AP) was decreased and firing rate were increased. VGSCs were up-regulated at both the mRNA and protein levels. The observed effects of TNF-α on Na(+) currents were inhibited by pre-incubation with the NF-κB inhibitor BAY 11-7082 (1 µM) or the p38 mitogen-activated protein kinases (MAPK) inhibitor SB203580 (1 µM). CONCLUSIONS: TNF-α increases Na(+) currents by accelerating the channel activation as well as increasing the expression of VGSCs in a mechanism dependent upon NF-κB and p38 MAPK signal pathways in CNS neurons.
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Córtex Cerebral/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Fator de Necrose Tumoral alfa/farmacologia , Canais de Sódio Disparados por Voltagem/efeitos dos fármacos , Canais de Sódio Disparados por Voltagem/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Células Cultivadas , Córtex Cerebral/citologia , Córtex Cerebral/fisiologia , Relação Dose-Resposta a Droga , Imidazóis/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , NF-kappa B/fisiologia , Neurônios/citologia , Nitrilas/farmacologia , Técnicas de Patch-Clamp , Piridinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Sulfonas/farmacologia , Fatores de Tempo , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologiaRESUMO
Anti-malaria drug, chloroquine, has been reported to be effective against influenza A virus (IAV) in vitro and used in in-vivo experiments and clinical trial for prevention or treatment of influenza. In this study, it has been shown by immunofluorescence, hemagglutination, and plaque assays that chloroquine enhanced A/WSN/33 (H1N1) replication with pronounced cytopathic effect in dose-, time-, and MOI-dependent manners in human lung epithelial cells A549. Time-of-addition assay showed that inhibitory effect on virus replication by chloroquine pre-treatment was indistinctive, and virus productions were enhanced when the drug was applied after viral adsorption. The effectiveness of chloroquine as an anti-influenza drug is questioned, and caution in its use is recommended.
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Cloroquina/farmacologia , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N1/fisiologia , Replicação Viral/efeitos dos fármacos , Antimaláricos/farmacologia , Antivirais/farmacologia , Linhagem Celular Tumoral , Células Cultivadas , Efeito Citopatogênico Viral/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Epiteliais/virologia , Humanos , Fatores de TempoRESUMO
AIMS: Increasing evidence suggests an association between diabetes and risk of venous thromboembolism (VTE); however, the results are inconsistent. We conducted a systematic review and meta-analysis of all epidemiological evidence to clarify association of diabetes with risk of VTE. METHODS: We searched MEDLINE and EMBASE to retrieve all relevant articles. Pooled effect estimates were calculated through a random-effects model. RESULTS: Sixteen articles involving 803,627,121 participants and 10,429,227 VTE patients were included. Pooled analysis of all evidence suggested that diabetes was associated with increased risk of VTE (HR, 1.35; 95%CI, 1.17-1.55; p=2.92*10(-5)), with evidence of small-study effect (p=0.024) and heterogeneity (I(2)=87.1%, p<0.001). However, when analysis was restricted to high quality cohort studies, the association remained significantly (HR, 1.36; 95%CI 1.11-1.68; p=0.004), with no evidence of publication bias (p=0.192) and heterogeneity (I(2)=23.2%, p=0.245). CONCLUSIONS: Diabetes is associated with increased risk of VTE, which may have implications for the primary and secondary prevention of VTE.
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Complicações do Diabetes , Diabetes Mellitus/terapia , Tromboembolia Venosa/complicações , Estudos de Coortes , Feminino , Humanos , Masculino , Fatores de Risco , Tamanho da Amostra , Resultado do TratamentoRESUMO
Macrophages polarized to M1 (pro-inflammation) or M2 (anti-inflammation) phenotypes in response to environmental signals. In this study, we examined the polarization of alveolar macrophage (AM), following induction by different influenza virus strains (ST169 (H1N1), ST602 (H3N2) and HKG9 (H9N2)). Macrophages from other tissues or cell line exert alternative responding pattern, and AM is necessary for investigating the respiratory system. AM polarized toward the M1 phenotype after 4 hours of infection by all three virus strains, and AM to presented M2b phenotype after 8 hours induction, and immunosuppressive phenotype after 24 hours of induction. Protein expression assay showed similar results as the gene expression analysis for phenotype verification. The ELISA assay showed that TNF-α secretion was up-regulated after 4 and 8 hours of infection by influenza viruses, and it returned to basal levels after 24 hours of infection. IL-10 expression was elevated after 8 and 24 hours of infection. Immunofluorescence showed that iNOS expression was up-regulated but not Arg1 expression. Influenza virus notably increased phospho-Akt but not phospho-Erk1/2 or phospho-p38, and the AM polarization pattern have been changed by LY294002 (PI3K inhibitor). In conclusion, our results demonstrate the dynamic polarization of AM induced by influenza viruses, and suggested that PI3K/Akt signaling pathway modulates AM polarization to M1/M2b.
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Vírus da Influenza A/imunologia , Macrófagos Alveolares/imunologia , Macrófagos Alveolares/virologia , Infecções por Orthomyxoviridae/imunologia , Fosfatidilinositol 3-Quinases/imunologia , Proteínas Proto-Oncogênicas c-akt/imunologia , Animais , Feminino , Humanos , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vírus da Influenza A Subtipo H9N2/imunologia , Influenza Humana/imunologia , Interleucina-10/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/imunologia , Infecções por Orthomyxoviridae/veterinária , Transdução de Sinais , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Typical antipsychotics are characterized by extrapyramidal syndrome (EPS). Previous studies demonstrated that typical antipsychotics could inhibit neuronal voltage-gated sodium channel (VGSC). However, EPS typically emerge only upon prolonged exposure. As a result, we examined effects of haloperidol, a prototype typical antipsychotic, on neuronal VGSC upon incubation for varying duration. Briefly, VGSC currents were activated and recorded using a whole-cell patch-clamp technique in primary culture of mouse cortical neurons. VGSC activity was inhibited by acute haloperidol exposure (for minutes), but enhanced in a time- and concentration-dependent manner by chronic haloperidol exposure (for hours). The effects of chronic haloperidol were associated with increased expression of VGSC subunits as well as corresponding electrophysiological channel properties. In summary, we found enhanced VGSC currents upon chronic haloperidol exposure in cortical neurons in contrast to inhibition by acute haloperidol exposure. Such a results may contribute to EPS of typical antipsychotics.
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Potenciais de Ação/fisiologia , Córtex Cerebral/fisiologia , Haloperidol/administração & dosagem , Ativação do Canal Iônico/fisiologia , Neurônios/fisiologia , Sódio/metabolismo , Canais de Sódio Disparados por Voltagem/metabolismo , Potenciais de Ação/efeitos dos fármacos , Animais , Antipsicóticos/administração & dosagem , Células Cultivadas , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Antagonistas de Dopamina/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/efeitos dos fármacosRESUMO
It has been reported that autophagy is involved in the replication of many viruses. In this study, we screened 89 medicinal plants, using an assay based on the inhibition of the formation of the Atg12-Atg5/Atg16 heterotrimer, an important regulator of autophagy, and selected Silybum marianum L. for further study. An antiviral assay indicated that silybin (S0), the major active compound of S. marianum L., can inhibit influenza A virus (IAV) infection. We later synthesized 5 silybin derivatives (S1 through S5) and found that 23-(S)-2-amino-3-phenylpropanoyl-silybin (S3) had the best activity. When we compared the polarities of the substituent groups, we found that the hydrophobicity of the substituent groups was positively correlated with their activities. We further studied the mechanisms of action of these compounds and determined that S0 and S3 also inhibited both the formation of the Atg12-Atg5/Atg16 heterotrimer and the elevated autophagy induced by IAV infection. In addition, we found that S0 and S3 could inhibit several components induced by IAV infection, including oxidative stress, the activation of extracellular signal-regulated kinase (ERK)/p38 mitogen-activated protein kinase (MAPK) and IκB kinase (IKK) pathways, and the expression of autophagic genes, especially Atg7 and Atg3. All of these components have been reported to be related to the formation of the Atg12-Atg5/Atg16 heterotrimer, which might validate our screening strategy. Finally, we demonstrated that S3 can significantly reduce influenza virus replication and the associated mortality in infected mice. In conclusion, we identified 23-(S)-2-amino-3-phenylpropanoyl-silybin as a promising inhibitor of IAV infection.
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Antivirais/farmacologia , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Extratos Vegetais/química , Silybum marianum/química , Silimarina/análogos & derivados , Animais , Antivirais/síntese química , Antivirais/isolamento & purificação , Autofagia/efeitos dos fármacos , Proteína 12 Relacionada à Autofagia , Proteína 5 Relacionada à Autofagia , Proteínas Relacionadas à Autofagia , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Chlorocebus aethiops , Cães , Regulação da Expressão Gênica , Ensaios de Triagem em Larga Escala , Humanos , Vírus da Influenza A Subtipo H1N1/crescimento & desenvolvimento , Células Madin Darby de Rim Canino , Proteínas Associadas aos Microtúbulos/antagonistas & inibidores , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Plasmídeos , Multimerização Proteica/efeitos dos fármacos , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Silimarina/síntese química , Silimarina/isolamento & purificação , Silimarina/farmacologia , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/antagonistas & inibidores , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/genética , Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina/metabolismo , Células VeroRESUMO
Autophagy is involved in many human diseases, such as cancer, cardiovascular disease and virus infection, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), influenza A virus (IAV) and coxsackievirus B3/B4 (CVB3/B4), so a drug screening model targeting autophagy may be very useful for the therapy of these diseases. In our study, we established a drug screening model based on the inhibition of the dissociation of Beclin1-Bcl2 heterodimer, an important negative regulator of autophagy, using bimolecular fluorescence complementation (BiFC) technique for developing novel autophagy inhibitors and anti-IAV agents. From 86 examples of traditional Chinese medicines, we found Syzygium aromaticum L. had the best activity. We then determined the anti-autophagy and anti-IAV activity of eugenol, the major active compound of Syzygium aromaticum L., and explored its mechanism of action. Eugenol could inhibit autophagy and IAV replication, inhibited the activation of ERK, p38MAPK and IKK/NF-κB signal pathways and antagonized the effects of the activators of these pathways. Eugenol also ameliorated the oxidative stress and inhibited the expressions of autophagic genes. We speculated that the mechanism underlying might be that eugenol inhibited the oxidative stress and the activation of ERK1/2, p38MAPK and IKK/NF-κB pathways, subsequently inhibited the dissociation of Beclin1-Bcl2 heterodimer and autophagy, and finally impaired IAV replication. These results might conversely display the reasonableness of the design of our screening model. In conclusion, we have established a drug screening model for developing novel autophagy inhibitor, and find eugenol as a promising inhibitor for autophagy and IAV infection.
Assuntos
Antivirais/farmacologia , Autofagia/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos/métodos , Eugenol/farmacologia , Vírus da Influenza A/efeitos dos fármacos , Linhagem Celular , Medicamentos de Ervas Chinesas/farmacologia , Humanos , Syzygium/químicaRESUMO
BCG-activated macrophages (BAM) could kill the tumor cells through cell-cell contact. In this process membrane proteins play an important role. However, up to date, few membrane proteins were revealed. In this study, we selected a surface molecule named Trim59, which was specifically expressed on BAM membrane (compared with the negative control). We cloned and prokaryoticly expressed the extracellular domain of Trim59, purified the recombinant protein and generated polyclonal antibodies. Immunohistochemistry showed that Trim59 abundantly expressed in spleen, stomach and ovary; intermediately expressed in brain, lung, kidney, muscle and intestine; but not in thymus, liver, heart, uterus. Using the antibodies to block Trim59 on BAM significantly reduced BAM cytotoxicity against MCA207 cells. This demonstrated that Trim59 serves as an indispensable molecule in maintaining BAM activity. Overexpression of Trim59 in Raw264.7 cell line failed to lyse target MCA207 cells, which potentiated Trim59 per se could not enhance macrophage cytotoxicity; on another hand, overexpression of Trim59 enhance the pinocytosis and Phagocytosis activity of Raw-264.7, which imply Trim59 might mediate the cell-molecule interaction. Our results indicate Trim59 might be an essential accessory molecule in mediating BAM tumoricidal functions; and Trim59 is a phagocytosis-correlated molecule.