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1.
Nat Genet ; 52(12): 1294-1302, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33077915

RESUMO

Angiotensin-converting enzyme 2 (ACE2) is an entry receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and a regulator of several physiological processes. ACE2 has recently been proposed to be interferon (IFN) inducible, suggesting that SARS-CoV-2 may exploit this phenomenon to enhance viral spread and questioning the efficacy of IFN treatment in coronavirus disease 2019. Using a recent de novo transcript assembly that captured previously unannotated transcripts, we describe a new isoform of ACE2, generated by co-option of intronic retroelements as promoter and alternative exon. The new transcript, termed MIRb-ACE2, exhibits specific expression patterns across the aerodigestive and gastrointestinal tracts and is highly responsive to IFN stimulation. In contrast, canonical ACE2 expression is unresponsive to IFN stimulation. Moreover, the MIRb-ACE2 translation product is a truncated, unstable ACE2 form, lacking domains required for SARS-CoV-2 binding and is therefore unlikely to contribute to or enhance viral infection.


Assuntos
/biossíntese , Interferons/metabolismo , Retroelementos/genética , /genética , Animais , Linhagem Celular , Chlorocebus aethiops , Indução Enzimática , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Regulação Viral da Expressão Gênica , Células HEK293 , Humanos , Isoenzimas/biossíntese , Isoenzimas/genética , Estabilidade Proteica , RNA-Seq , /metabolismo , Distribuição Tecidual , Células Vero
2.
Nat Genet ; 52(12): 1283-1293, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33077916

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes COVID-19, utilizes angiotensin-converting enzyme 2 (ACE2) for entry into target cells. ACE2 has been proposed as an interferon-stimulated gene (ISG). Thus, interferon-induced variability in ACE2 expression levels could be important for susceptibility to COVID-19 or its outcomes. Here, we report the discovery of a novel, transcriptionally independent truncated isoform of ACE2, which we designate as deltaACE2 (dACE2). We demonstrate that dACE2, but not ACE2, is an ISG. In The Cancer Genome Atlas, the expression of dACE2 was enriched in squamous tumors of the respiratory, gastrointestinal and urogenital tracts. In vitro, dACE2, which lacks 356 amino-terminal amino acids, was non-functional in binding the SARS-CoV-2 spike protein and as a carboxypeptidase. Our results suggest that the ISG-type induction of dACE2 in IFN-high conditions created by treatments, an inflammatory tumor microenvironment or viral co-infections is unlikely to increase the cellular entry of SARS-CoV-2 and promote infection.


Assuntos
/metabolismo , Interferons/metabolismo , Vírus de RNA/fisiologia , /metabolismo , /química , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/genética , Linhagem Celular , Indução Enzimática , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Humanos , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Mucosa Respiratória/metabolismo , Mucosa Respiratória/virologia , Glicoproteína da Espícula de Coronavírus/metabolismo
3.
Neurologia ; 35(9): 628-632, 2020.
Artigo em Inglês, Espanhol | MEDLINE | ID: mdl-32896463

RESUMO

INTRODUCTION: In recent months, doubts have arisen among patients, general practitioners, and neurologists as to whether some drugs commonly used in patients with headaches and neuralgia may favour or complicate the disease caused by SARS-CoV-2. MATERIAL AND METHODS: We collected information on the opinions of scientific societies and medicines agencies (American, European, and Spanish) to clarify doubts regarding the use of drugs such as lisinopril, candesartan, ibuprofen, corticosteroids, carbamazepine, and monoclonal antibodies targeting the calcitonin gene-related peptide in the context of the COVID-19 pandemic. RESULTS: We make recommendations about the use of standard headache treatments in the context of the COVID-19 pandemic, based on the current scientific evidence. CONCLUSIONS: At present, there is no robust scientific argument to formally contraindicate any of the standard treatments employed for headaches and neuralgias.


Assuntos
Analgésicos/efeitos adversos , Infecções por Coronavirus/complicações , Cefaleia/tratamento farmacológico , Neuralgia/tratamento farmacológico , Pneumonia Viral/complicações , Corticosteroides/efeitos adversos , Corticosteroides/uso terapêutico , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais/uso terapêutico , Anticonvulsivantes/efeitos adversos , Anticonvulsivantes/uso terapêutico , Anti-Hipertensivos/efeitos adversos , Anti-Hipertensivos/uso terapêutico , Antivirais/farmacologia , Benzimidazóis/efeitos adversos , Benzimidazóis/uso terapêutico , Betacoronavirus , Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina/efeitos adversos , Antagonistas do Receptor do Peptídeo Relacionado ao Gene de Calcitonina/uso terapêutico , Carbamazepina/efeitos adversos , Carbamazepina/uso terapêutico , Infecções por Coronavirus/tratamento farmacológico , Suscetibilidade a Doenças/induzido quimicamente , Interações Medicamentosas , Indução Enzimática/efeitos dos fármacos , Cefaleia/complicações , Cefaleia/prevenção & controle , Humanos , Ibuprofeno/efeitos adversos , Ibuprofeno/farmacologia , Ibuprofeno/uso terapêutico , Lisinopril/efeitos adversos , Lisinopril/uso terapêutico , Neuralgia/complicações , Pandemias , Peptidil Dipeptidase A/biossíntese , Peptidil Dipeptidase A/genética , Receptores Virais/biossíntese , Receptores Virais/genética , Fatores de Risco , Tetrazóis/efeitos adversos , Tetrazóis/uso terapêutico
4.
Mol Pharmacol ; 98(5): 634-647, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32892155

RESUMO

Long-term administration of some antiepileptic drugs often increases blood lipid levels. In this study, we investigated its molecular mechanism by focusing on the nuclear receptors constitutive active/androstane receptor (CAR) and peroxisome proliferator-activated receptor α (PPARα), which are key transcription factors for enzyme induction and lipid metabolism, respectively, in the liver. Treatment of mice with the CAR activator phenobarbital, an antiepileptic drug, increased plasma triglyceride levels and decreased the hepatic expression of PPARα target genes related to lipid metabolism. The increase in PPARα target gene expression induced by fenofibrate, a PPARα ligand, was inhibited by cotreatment with phenobarbital. CAR suppressed PPARα-dependent gene transcription in HepG2 cells but not in COS-1 cells. The mRNA level of peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α), a coactivator for both CAR and PPARα, in COS-1 cells was much lower than in HepG2 cells. In reporter assays with COS-1 cells overexpressing PGC1α, CAR suppressed PPARα-dependent gene transcription, depending on the coactivator-binding motif. In mammalian two-hybrid assays, CAR attenuated the interaction between PGC1α and PPARα Chemical inhibition of PGC1α prevented phenobarbital-dependent increases in plasma triglyceride levels and the inhibition of PPARα target gene expression. These results suggest that CAR inhibits the interaction between PPARα and PGC1α, attenuating PPARα-dependent lipid metabolism. This might explain the antiepileptic drug-induced elevation of blood triglyceride levels. SIGNIFICANCE STATEMENT: Constitutive active/androstane receptor activated by antiepileptic drugs inhibits the peroxisome proliferator-activated receptor α-dependent transcription of genes related to lipid metabolism and upregulates blood triglyceride levels. The molecular mechanism of this inhibition involves competition between these nuclear receptors for coactivator peroxisome proliferator-activated receptor γ coactivator-1α binding.


Assuntos
Anticonvulsivantes/farmacologia , PPAR alfa/metabolismo , PPAR gama/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Triglicerídeos/sangue , Animais , Linhagem Celular Tumoral , Indução Enzimática/efeitos dos fármacos , Fenofibrato/farmacologia , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fenobarbital/farmacologia , Fatores de Transcrição/metabolismo , Ativação Transcricional/efeitos dos fármacos
5.
Sci Rep ; 10(1): 15244, 2020 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-32943672

RESUMO

Tamoxifen (TAM) inducible Cre recombinase system is an essential tool to study gene function when early ablation or overexpression can cause developmental defects or embryonic lethality. However, there remains a lack of consensus on the optimal route and dosage of TAM administration in vivo. Here, we assessed dosage and delivery of TAM for activation of Cre in immune cell subsets assessed longitudinally and spatially using transgenic mice with ubiquitously expressed Cre/ER and the Cre-inducible fluorescent reporter YFP. After comparing two TAM delivery methods (intraperitoneal versus oral gavage) and different doses, we found that 3 mg of TAM administered orally for five consecutive days provides maximal reporter induction with minimal adverse effects in vivo. Serum levels of TAM peaked 1 week after initiating treatment then slowly decreased, regardless of dosing and delivery methods. TAM concentration in specific tissues (liver, spleen, lymph nodes, and thymus) was also dependent on delivery method and dose. Cre induction was highest in myeloid cells and B cells and substantially lower in T cells, and double-positive thymocytes had a notably higher response to TAM. In addition to establishing optimal dose and administration of TAM, our study reveals a disparate activity of Cre in different cell immune populations when using Cre/ER models.


Assuntos
Sistema Imunitário/citologia , Sistema Imunitário/enzimologia , Integrases/biossíntese , Tamoxifeno/farmacologia , Administração Oral , Animais , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Relação Dose-Resposta a Droga , Indução Enzimática/efeitos dos fármacos , Feminino , Genes Reporter , Sistema Imunitário/efeitos dos fármacos , Injeções Intraperitoneais , Integrases/genética , Antígenos Comuns de Leucócito/metabolismo , Fígado/citologia , Fígado/efeitos dos fármacos , Fígado/imunologia , Proteínas Luminescentes/biossíntese , Proteínas Luminescentes/genética , Tecido Linfoide/citologia , Tecido Linfoide/efeitos dos fármacos , Tecido Linfoide/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Tamoxifeno/administração & dosagem , Tamoxifeno/farmacocinética
6.
Sci Rep ; 10(1): 13978, 2020 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-32814815

RESUMO

We report that the naphthalimide analogue 2-(2-aminophenyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (NAP-6) is a highly potent and selective breast cancer targeting molecule. These effects are mediated via the aryl hydrocarbon receptor (AHR) pathway and the subsequent induction of CYP1 metabolising monooxygenases in breast cancer cell line models. Indeed the triple negative breast cancer cell line MDA-MB-468 with a GI50 value of 100 nM is greater than 500-fold more sensitive to NAP-6 compared with other tumour derived cell models. Within 1 h exposure of these cells to NAP-6, CYP1A1 expression increases 25-fold, rising to 250-fold by 24 h. A smaller concurrent increase in CYP1A2 and CYP1B1 is also observed. Within 24 h these cells present with DNA damage as evident by enhanced H2AXγ expression, cell cycle checkpoint activation via increased CHK2 expression, S-phase cell cycle arrest and cell death. Specific small molecule inhibitors of the AHR and CYP1 family ameliorate these events. A positive luciferase reporter assay for NAP-6 induced XRE binding further confirms the role of the AHR in this phenomenon. Non-sensitive cell lines fail to show these biological effects. For the first time we identify 2-(2-aminophenyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione as a new AHR ligand that selectively targets breast cancer.


Assuntos
Neoplasias da Mama/metabolismo , Naftalimidas/farmacologia , Receptores de Hidrocarboneto Arílico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A2/genética , Citocromo P-450 CYP1A2/metabolismo , Citocromo P-450 CYP1B1/genética , Citocromo P-450 CYP1B1/metabolismo , Dano ao DNA , Indução Enzimática/efeitos dos fármacos , Feminino , Células HT29 , Humanos , Células MCF-7 , Estrutura Molecular , Naftalimidas/química , Receptores de Hidrocarboneto Arílico/genética , Transdução de Sinais/genética , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia
7.
Sci Rep ; 10(1): 14072, 2020 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-32826925

RESUMO

Altered metabolism of fatty acid synthesis is considered a hallmark characteristic of several malignancies, including acute lymphoblastic leukemia (ALL). To evaluate the impact of fatty acid synthase (FASN) on drug resistant ALL, bone marrow samples were collected from 65 pediatric ALLs, including 40 de novo and 25 relapsed patients. 22 non-cancer individuals were chosen as controls. Quantitative RT-PCR showed increased expression levels of FASN in drug resistant patients compared with the therapy responders. Single and combined treatment of malignant cells were analyzed using Annexin-V/PI double staining and MTT assays. Incubation of resistant primary cells with ginger showed simultaneous increased apoptosis rates and reduced FASN expression levels. Furthermore, docking studies demonstrated high affinity bindings between ginger derivatives and FASN thioesterase and ketosynthase domains, compared with their known inhibitors, fenofibrate and morin, respectively. Finally, combined treatment of in-house multidrug resistant T-ALL subline with ginger and dexamethasone induced drug sensitivity and down regulation of FASN expression, accordingly. To the best of our knowledge, this is the first study that introduces FASN upregulation as a poor prognostic factor for drug resistant childhood ALL. Moreover, it was revealed that FASN inhibition may be applied by ginger phytochemicals and overcome dexamethasone resistance, subsequently.


Assuntos
Ácido Graxo Sintase Tipo I/antagonistas & inibidores , Gengibre/química , Terapia de Alvo Molecular , Proteínas de Neoplasias/antagonistas & inibidores , Extratos Vegetais/uso terapêutico , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Apoptose/efeitos dos fármacos , Medula Óssea/enzimologia , Estudos de Casos e Controles , Criança , Dexametasona/farmacologia , Dexametasona/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Indução Enzimática/efeitos dos fármacos , Feminino , Fenofibrato/farmacologia , Flavonoides/farmacologia , Regulação Leucêmica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Modelos Moleculares , Simulação de Acoplamento Molecular , Extratos Vegetais/farmacologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/enzimologia , Prognóstico , Conformação Proteica , Domínios Proteicos , RNA Mensageiro/biossíntese , RNA Neoplásico/biossíntese , Células Tumorais Cultivadas
8.
Sci Rep ; 10(1): 14164, 2020 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-32843697

RESUMO

Activation-induced cytidine deaminase (AID) is one kind of the mutant enzymes, which target regulating the immunoglobulin (Ig) gene in Burkitt's lymphoma to initiate class switch recombination (CSR), resulting in c-Myc chromosomal translocation. However, it is not clear that whether AID induces c-Myc/IgH translocation in double-hit lymphoma (DHL) with c-Myc gene translocation. In this study, the AID in DHL tissues and classical diffuse large b-cell lymphoma (DLBCL) tissues were compared. The results suggested that AID is of important value in predicting DHL, stronger CSR of AID was observed in DHL patients, which exhibited AID overexpression and c-Myc gene translocation of DHL after CSR induction. It is concluded that AID directly induces CSR in DHL and may result in c-Myc gene translocation. Targeting AID may be a good treatment regimen for DHL.


Assuntos
Citidina Desaminase/biossíntese , Switching de Imunoglobulina/genética , Linfoma Difuso de Grandes Células B/enzimologia , Terapia de Alvo Molecular , Proteínas de Neoplasias/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Citidina Desaminase/genética , Citidina Desaminase/fisiologia , Indução Enzimática/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes bcl-2 , Genes myc , Humanos , Isotipos de Imunoglobulinas/biossíntese , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/genética , Estimativa de Kaplan-Meier , Antígeno Ki-67/genética , Lipopolissacarídeos/farmacologia , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/mortalidade , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/fisiologia , Prognóstico , Proteínas Proto-Oncogênicas c-bcl-6/genética , Translocação Genética , Regulação para Cima/efeitos dos fármacos
9.
Vascul Pharmacol ; 133-134: 106777, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32750408

RESUMO

Atherosclerosis is a systemic chronic inflammatory disease. Many antioxidants including alpha-lipoic acid (LA), a product of lipoic acid synthase (Lias), have proven to be effective for treatment of this disease. However, the question remains whether LA regulates the immune response as a protective mechanism against atherosclerosis. We initially investigated whether enhanced endogenous antioxidant can retard the development of atherosclerosis via immunomodulation. To explore the impact of enhanced endogenous antioxidant on the retardation of atherosclerosis via immune regulation, our laboratory has recently created a double mutant mouse model, using apolipoprotein E-deficient (Apoe-/-) mice crossbred with mice overexpressing lipoic acid synthase gene (LiasH/H), designated as LiasH/HApoe-/- mice. Their littermates, Lias+/+Apoe-/- mice, served as a control. Distinct redox environments between the two strains of mice have been established and they can be used to facilitate identification of antioxidant targets in the immune response. At 6 months of age, LiasH/HApoe-/- mice had profoundly decreased atherosclerotic lesion size in the aortic sinus compared to their Lias+/+Apoe-/- littermates, accompanied by significantly enhanced numbers of regulatory T cells (Tregs) and anti-oxidized LDL autoantibody in the vascular system, and reduced T cell infiltrates in aortic walls. Our results represent a novel exploration into an environment with increased endogenous antioxidant and its ability to alleviate atherosclerosis, likely through regulation of the immune response. These outcomes shed light on a new therapeutic strategy using antioxidants to lessen atherosclerosis.


Assuntos
Aorta/enzimologia , Doenças da Aorta/prevenção & controle , Aterosclerose/prevenção & controle , Placa Aterosclerótica , Sulfurtransferases/biossíntese , Animais , Aorta/imunologia , Aorta/patologia , Doenças da Aorta/enzimologia , Doenças da Aorta/imunologia , Doenças da Aorta/patologia , Aterosclerose/enzimologia , Aterosclerose/imunologia , Aterosclerose/patologia , Autoanticorpos/sangue , Modelos Animais de Doenças , Indução Enzimática , Lipoproteínas LDL/imunologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout para ApoE , Oxirredução , Estresse Oxidativo , Sulfurtransferases/genética , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo
10.
Med Hypotheses ; 143: 110112, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32721806

RESUMO

In coronavirus disease-19 (COVID-19), four major factors have been correlated with worse prognosis: aging, hypertension, obesity, and exposure to androgen hormones. Angiotensin-converting enzyme-2 (ACE2) receptor, regulation of the renin-angiotensin-aldosterone system (RAAS), and transmembrane serine protease 2 (TMPRSS2) action are critical for the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) cell entry and infectivity. ACE2 expression and RAAS are abnormal in hypertension and obesity, while TMPRSS2 is overexpressed when exposed to androgens, which may justify why these factors are overrepresented in COVID-19. Among therapeutic targets for SARS-CoV-2, we hypothesized that spironolactone, a long used and safe mineralocorticoid and androgen receptors antagonist, with effective anti-hypertensive, cardioprotective, nephroprotective, and anti-androgenic properties may offer pleiotropic actions in different sites to protect from COVID-19. Current data shows that spironolactone may concurrently mitigate abnormal ACE2 expression, correct the balances membrane-attached and free circulating ACE2 and between angiotensin II and Angiotensin-(1-7) (Ang-(1-7)), suppress androgen-mediated TMPRSS2 activity, and inhibit obesity-related RAAS dysfunctions, with consequent decrease of viral priming. Hence, spironolactone may provide protection from SARS-CoV-2, and has sufficient plausibility to be clinically tested, particularly in the early stages of COVID-19.


Assuntos
Antagonistas de Androgênios/uso terapêutico , Androgênios/fisiologia , Betacoronavirus/fisiologia , Infecções por Coronavirus/tratamento farmacológico , Antagonistas de Receptores de Mineralocorticoides/uso terapêutico , Pandemias , Pneumonia Viral/tratamento farmacológico , Sistema Renina-Angiotensina/efeitos dos fármacos , Espironolactona/uso terapêutico , Antagonistas de Androgênios/farmacologia , Inibidores da Enzima Conversora de Angiotensina/uso terapêutico , Cardiotônicos/farmacologia , Cardiotônicos/uso terapêutico , Infecções por Coronavirus/complicações , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/fisiopatologia , Indução Enzimática/efeitos dos fármacos , Humanos , Hipertensão/complicações , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Rim/efeitos dos fármacos , Masculino , Antagonistas de Receptores de Mineralocorticoides/farmacologia , Obesidade/complicações , Obesidade/fisiopatologia , Peptidil Dipeptidase A/biossíntese , Peptidil Dipeptidase A/efeitos dos fármacos , Pneumonia Viral/complicações , Pneumonia Viral/epidemiologia , Pneumonia Viral/fisiopatologia , Prognóstico , Receptores Virais/efeitos dos fármacos , Fatores de Risco , Serina Endopeptidases/efeitos dos fármacos , Distribuição por Sexo , Espironolactona/farmacologia , Internalização do Vírus/efeitos dos fármacos
11.
Sci Rep ; 10(1): 10885, 2020 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-32616748

RESUMO

Ten distinct cDNAs encoding five different protein phosphatases 1 (PPP1) were cloned from Nilaparvata lugens. NlPPP1α and NlPPP1ß are highly conserved whereas NlPPP1-Y, NlPPP1-Y1 and NlPPP1-Y2 are lowly conserved among insects. NlPPP1α and NlPPP1ß exhibited a ubiquitous expression, while NlPPP1-Y, NlPPP1-Y1, and NlPPP1-Y2 were obviously detected from the 4th instar nymph to imago developmental stages in males, especially detected in internal reproductive organ and fat bodies of the male. Injection nymphs with dsRNA of NlPPP1α or NlPPP1ß was able to reduce the target gene expression in a range of 71.5-91.0%, inducing a maximum mortality rate of 95.2% or 97.2% at 10th day after injection and eclosion ratio down by 65.5-100.0%. Injection with dsNlPPP1Ys targeted to NlPPP1-Y, NlPPP1-Y1and NlPPP1-Y2 was able to induce a maximum mortality rate of 95.5% at 10th day after injection, eclosion ratio down by 86.4%. Knock-down one of the male-biased NlPPP1 genes has no effect on survival and eclosion ratio. Injection of 4th instar nymph with dsNlPPP1Ys led to reduced oviposition amount and hatchability, down by 44.7% and 19.6% respectively. Knock-down of NlPPP1-Y1 or NlPPP1-Y2 gene did not significantly affect oviposition amount but significantly affected hatchability. The results indicate that the male-biased NlPPP1 genes have overlapping functions in N. lugens development, and NlPPP1-Y1 and NlPPP1-Y2 may play important roles in spermatogenesis and fertilization. The dsNlPPP1ß and dsNlPPP1Ys in this study could be the preferred sequence in RNAi and low-conserved male-biased NlPPP1 genes could be potential target for N. lugens control.


Assuntos
Genes de Insetos , Hemípteros/genética , Proteína Fosfatase 1/genética , Regiões 5' não Traduzidas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Sequência Conservada , DNA Complementar/genética , Indução Enzimática , Feminino , Fertilidade/efeitos dos fármacos , Hemípteros/enzimologia , Hemípteros/crescimento & desenvolvimento , Masculino , Oócitos/ultraestrutura , Especificidade de Órgãos , Fosforilação , Filogenia , Isoformas de Proteínas/genética , Isoformas de Proteínas/toxicidade , Proteína Fosfatase 1/toxicidade , Processamento de Proteína Pós-Traducional , Subunidades Proteicas , RNA/genética , Interferência de RNA , Alinhamento de Sequência , Homologia de Sequência , Ducto Deferente/anormalidades
12.
Sci Rep ; 10(1): 10879, 2020 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-32616893

RESUMO

Although the severity of influenza virus infections has been associated with host energy metabolism, the related mechanisms have not yet been clarified. Here we examined the effects of influenza virus infection on host energy metabolism in mice. After infecting mice with intranasal applications of 500 plaque-forming units of A/Puerto Rico/8/34 (H1N1; PR8) virus, the serum levels of most intermediates in the tricarboxylic acid (TCA) cycle and related metabolic pathways were significantly reduced. These data suggest that substrate supply to the TCA cycle is reduced under these conditions, rather than specific metabolic reactions being inhibited. Then, we focused on glucose and fatty acid metabolism that supply substrates to the TCA cycle. Akt phosphorylation following insulin injections was attenuated in the livers of PR8 virus-infected mice. Furthermore, glucose tolerance tests revealed that the PR8 virus-infected mice showed higher blood glucose levels than the vehicle-inoculated control mice. These results suggest that influenza virus infection impairs insulin signaling, which regulates glucose uptake. However, increases in the hepatic expressions of fatty acid-metabolizing enzymes suggest that fatty acids accumulate in liver cells of infected mice. Collectively, our data indicate that influenza virus infection dysregulates host energy metabolism. This line of investigation provides novel insights into the pathogenesis of influenza.


Assuntos
Ciclo do Ácido Cítrico , Metabolismo Energético , Ácidos Graxos/metabolismo , Vírus da Influenza A Subtipo H1N1/fisiologia , Insulina/fisiologia , Infecções por Orthomyxoviridae/metabolismo , Animais , Citocinas/sangue , Indução Enzimática , Regulação da Expressão Gênica , Glucose/metabolismo , Resistência à Insulina , Fígado/metabolismo , Fígado/virologia , Masculino , Metaboloma , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação , Processamento de Proteína Pós-Traducional , Proteínas Proto-Oncogênicas c-akt/metabolismo , Purinas/metabolismo , Transdução de Sinais
13.
Ecotoxicol Environ Saf ; 203: 110997, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32684518

RESUMO

A novel study on biodegradation of 30 mg L-1 of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) mixture (celecoxib, diclofenac and ibuprofen) by two wood-rot fungi; Ganoderma applanatum (GA) and Laetiporus sulphureus (LS) was investigated for 72 h. The removal efficiency of celecoxib, diclofenac and ibuprofen were 98, 96 and 95% by the fungal consortium (GA + LS). Although, both GA and LS exhibited low removal efficiency (61 and 73% respectively) on NSAIDs. However, 99.5% degradation of the drug mixture (NSAIDs) was achieved on the addition of the fungal consortium (GA + LS) to the experimental set-up. Overall, LS exhibited higher degradation efficiency; 92, 87, 79% on celecoxib, diclofenac and ibuprofen than GA with 89, 80 and 66% respectively. Enzyme analyses revealed significant induction of 201, 180 and 135% in laccase (Lac), lignin peroxidase (LiP) and manganese peroxidase (MnP) by the fungal consortium during degradation of the NSAIDs respectively. The experimental data showed the best goodness of fit when subjected to Langmuir (R2 = 0.980) and Temkin (R2 = 0.979) isotherm models which suggests monolayer and heterogeneous nature exhibited by the mycelia during interactions with NSAIDs. The degradation mechanism followed pseudo-second-order kinetic model (R2 = 0.987) indicating the strong influence of fungal biomass in the degradation of NSAIDs. Furthermore, Gas Chromatography-Mass Spectrometry (GCMS) and High-Performance Liquid Chromatography (HPLC) analyses confirmed the degraded metabolic states of the NSAIDs after treatment with GA, LS and consortium (GA + LS). Hence, the complete removal of NSAIDs is best achieved in an economical and eco-friendly way with the use of fungi consortium.


Assuntos
Anti-Inflamatórios não Esteroides/análise , Poluentes Ambientais/análise , Ganoderma/enzimologia , Ganoderma/crescimento & desenvolvimento , Lignina/metabolismo , Madeira/microbiologia , Anti-Inflamatórios não Esteroides/metabolismo , Biodegradação Ambiental , Biomassa , Poluentes Ambientais/metabolismo , Indução Enzimática/efeitos dos fármacos , Cinética , Lacase/biossíntese , Modelos Biológicos , Peroxidases/biossíntese
14.
Am J Physiol Heart Circ Physiol ; 319(2): H341-H348, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32618512

RESUMO

Progesterone exerts antihypertensive actions partially by modulating endothelial nitric oxide synthase (eNOS) activity. Here, we aimed to investigate the effects and mechanisms of progesterone on eNOS expression. First, human umbilical vein endothelial cells (HUVECs) were exposed to progesterone and then the eNOS transcription factor specificity protein-1 (SP-1) and progesterone receptor (PRA/B) expression were assessed by Western blotting and qRT-PCR. The interaction between SP-1 and PRA/B was next determined through coimmunoprecipitation assay. The chromatin immunoprecipitation assay and luciferase assay were used to investigate the relationship of PRA/B, SP-1, and eNOS promoter. At last, rats were intraperitoneally injected with progesterone receptor antagonist RU-486, and then the expression of eNOS and vasodilation function in thoracic aorta and mesenteric artery were measured. The results showed that progesterone could increase eNOS expression in HUVECs. Further study showed that progesterone increased PRA-SP-1 complex formation and facilitated PRA/B and SP-1 binding to eNOS promoter. Mutating SP-1 or PR-binding motif on eNOS promoter abolished the effect of progesterone on eNOS gene transcription. We also observed that progesterone receptor antagonist RU-486 reduced eNOS expression and impaired vasodilation in rats. Those results suggest that progesterone modulates eNOS expression through promoting PRA-SP-1 complex formation, and progesterone antagonist attenuates eNOS expression, leading to the loss of vascular relaxation.NEW & NOTEWORTHY Progesterone directly upregulated endothelial nitric oxide synthase (eNOS) expression in human endothelial cells. Progesterone augmented eNOS promoter activity through a progesterone receptor A- and specificity protein-1-dependent manner. Antagonism of the progesterone receptor reduced eNOS expression and impaired vasodilation in rats.


Assuntos
Núcleo Celular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Óxido Nítrico Sintase Tipo III/biossíntese , Progesterona/farmacologia , Receptores de Progesterona/agonistas , Fator de Transcrição Sp1/metabolismo , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/enzimologia , Sítios de Ligação , Núcleo Celular/metabolismo , Células Cultivadas , Indução Enzimática , Feminino , Antagonistas de Hormônios/farmacologia , Células Endoteliais da Veia Umbilical Humana/enzimologia , Humanos , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/enzimologia , Óxido Nítrico Sintase Tipo III/genética , Regiões Promotoras Genéticas , Ratos Sprague-Dawley , Receptores de Progesterona/antagonistas & inibidores , Receptores de Progesterona/metabolismo , Transdução de Sinais , Vasodilatação/efeitos dos fármacos
15.
Sci Rep ; 10(1): 9575, 2020 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-32533042

RESUMO

To evaluate the effect and mechanism of radiotherapy (RT)-sorafenib pharmacokinetics (PK) in different regimens with conventional or high dose irradiation. Between February 2012 and December 2018, 43 patients with portal vein tumor thrombosis treated with sorafenib plus conventional RT (58%) or stereotactic body radiation therapy (SBRT, 42%) were retrospectively reviewed. In vivo and in vitro studies of concurrent and sequential RT with sorafenib were designed. SBRT resulted in a 3-fold increase in complete recanalization compared to conventional RT group (28% vs. 8%, p = 0.014). Compared to the control group, the area under the concentration vs. time curve (AUC) of sorafenib was increased in the concurrent RT2Gy and RT9Gy groups and the sequential RT9Gy group by 132% (p = 0.046), 163% (p = 0.038) and 102% (p = 0.018), respectively; and was decreased by 59% in the sequential RT2Gy group (p = 0.036). Sequential RT2Gy and RT9Gy increased CYP3A4 activity by 82% (p = 0.028) and 203% (p = 0.0004), respectively, compared to that with the corresponding concurrent regimen. SBRT produced better recanalization than conventional RT with sorafenib. The AUC of sorafenib was modulated by RT. P-gp expression was not influenced by RT. The sequential RT regimen increased CYP3A4 activity that may increase the RT-sorafenib synergy effect and overall sorafenib activity. The biodistribution of sorafenib was modulated by local RT with the different regimens.


Assuntos
Antineoplásicos/farmacocinética , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Veia Porta/efeitos da radiação , Inibidores de Proteínas Quinases/farmacocinética , Radiocirurgia/métodos , Sorafenibe/farmacocinética , Trombose Venosa/radioterapia , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/efeitos da radiação , Animais , Antineoplásicos/uso terapêutico , Carcinoma Hepatocelular/complicações , Linhagem Celular Tumoral , Terapia Combinada , Ciclosporina/farmacologia , Citocromo P-450 CYP3A/metabolismo , Inibidores do Citocromo P-450 CYP3A/farmacologia , Relação Dose-Resposta à Radiação , Indução Enzimática/efeitos da radiação , Humanos , Neoplasias Hepáticas/complicações , Masculino , NF-kappa B/metabolismo , NF-kappa B/efeitos da radiação , Inibidores de Proteínas Quinases/uso terapêutico , Ratos , Ratos Sprague-Dawley , Estudos Retrospectivos , Sorafenibe/uso terapêutico , Organismos Livres de Patógenos Específicos , Distribuição Tecidual , Trombose Venosa/etiologia
16.
Planta Med ; 86(12): 867-875, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32557519

RESUMO

Artemisia annua tea is a popular dosage form used to treat and prevent malaria in some developing countries. However, repeated drinking leads to an obviously decreased efficacy, which may be related to the induction of metabolizing enzymes by artemisinin. In the present study, the ability of different components in A. annua to activate the pregnane X receptor and constitutive androstane receptor was evaluated by the dual luciferase reporter gene system. The changes in mRNA and protein expression of CYP3A4 and CYP2B6 were determined by quantitative real-time PCR and Western blotting. Results showed that in the pregnane X receptor-mediated CYP3A4 reporter gene system, chrysosplenetin and arteannuin B exhibited a weak induction effect on pregnane X receptor wt, while arteannuin A had a strong induction effect on pregnane X receptor wt and pregnane X receptor 370 and a weak induction effect on pregnane X receptor 163. In the pregnane X receptor-mediated CYP2B6 reporter gene system, arteannuin A had a moderate induction effect on pregnane X receptor wt and pregnane X receptor 379, and a weak induction effect on pregnane X receptor 403, while arteannuin B had a weak induction effect on pregnane X receptor wt and pregnane X receptor 379. Arteannuin A had a strong induction effect on constitutive androstane receptor 3 in constitutive androstane receptor-mediated CYP3A4/2B6 reporter gene systems, while arteannuin B showed a weak induction effect on constitutive androstane receptor 3 in the constitutive androstane receptor-mediated CYP2B6 reporter gene system. The mRNA and protein expressions of CYP3A4 and CYP2B6 were increased when the pregnane X receptor or constitutive androstane receptor was activated. Various components present in A. annua differentially affect the activities of pregnane X receptor isoforms and the constitutive androstane receptor, which indicates the possibility of a drug-drug interaction. This partly explains the decline in efficacy after repeated drinking of A. annua tea.


Assuntos
Artemisia annua , Receptores de Esteroides/genética , Citocromo P-450 CYP2B6 , Citocromo P-450 CYP3A , Indução Enzimática , Hepatócitos , Extratos Vegetais , Receptores Citoplasmáticos e Nucleares
17.
Sci Rep ; 10(1): 7128, 2020 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-32346014

RESUMO

Acid-resistance systems are essential for pathogenic Escherichia coli to survive in the strongly acidic environment of the human stomach (pH < 2.5). Among these, the glutamic acid decarboxylase (GAD) system is the most effective. However, the precise mechanism of GAD induction is unknown. We previously reported that a tolC mutant lacking the TolC outer membrane channel was defective in GAD induction. Here, we show that indole, a substrate of TolC-dependent efflux pumps and produced by the tryptophanase encoded by the tnaA gene, negatively regulates GAD expression. GAD expression was restored by deleting tnaA in the tolC mutant; in wild-type E. coli, it was suppressed by adding indole to the growth medium. RNA-sequencing revealed that tnaA mRNA levels drastically decreased upon exposure to moderately acidic conditions (pH 5.5). This decrease was suppressed by RNase E deficiency. Collectively, our results demonstrate that the RNase E-dependent degradation of tnaA mRNA is accelerated upon acid exposure, which decreases intracellular indole concentrations and triggers GAD induction.


Assuntos
Endorribonucleases/metabolismo , Proteínas de Escherichia coli/genética , Escherichia coli/efeitos dos fármacos , Ácido Gástrico , RNA Mensageiro/metabolismo , Triptofanase/genética , Meios de Cultura , Indução Enzimática , Escherichia coli/enzimologia , Escherichia coli/genética , Glutamato Descarboxilase/biossíntese , Glutamato Descarboxilase/metabolismo , Hidrólise , Indóis/metabolismo
18.
Am J Physiol Renal Physiol ; 318(4): F1030-F1040, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32150446

RESUMO

Manipulation of circulating histidine-containing dipeptides (HCD) has been shown to affect the development of diabetes and early-stage diabetic nephropathy (DN). The aim of the present study was to investigate whether such interventions, which potentially alter levels of circulating HCD, also affect the development of advanced-stage DN. Two interventions, aerobic exercise training and overexpression of the human carnosinase-1 (hCN1) enzyme, were tested. BTBR ob/ob mice were either subjected to aerobic exercise training (20 wk) or genetically manipulated to overexpress hCN1, and different diabetes- and DN-related markers were compared with control ob/ob and healthy (wild-type) mice. An acute exercise study was performed to elucidate the effect of obesity, acute running, and hCN1 overexpression on plasma HCD levels. Chronic aerobic exercise training did not affect the development of diabetes or DN, but hCN1 overexpression accelerated hyperlipidemia and aggravated the development of albuminuria, mesangial matrix expansion, and glomerular hypertrophy of ob/ob mice. In line, plasma, kidney, and muscle HCD were markedly lower in ob/ob versus wild-type mice, and plasma and kidney HCD in particular were lower in ob/ob hCN1 versus ob/ob mice but were unaffected by aerobic exercise. In conclusion, advanced glomerular damage is accelerated in mice overexpressing the hCN1 enzyme but not protected by chronic exercise training. Interestingly, we showed, for the first time, that the development of DN is closely linked to renal HCD availability. Further research will have to elucidate whether the stimulation of renal HCD levels can be a therapeutic strategy to reduce the risk for developing DN.


Assuntos
Nefropatias Diabéticas/enzimologia , Dipeptidases/biossíntese , Terapia por Exercício , Glomérulos Renais/enzimologia , Músculo Esquelético/enzimologia , Obesidade/enzimologia , Animais , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/patologia , Dipeptidases/genética , Dipeptídeos/metabolismo , Modelos Animais de Doenças , Indução Enzimática , Histidina/análogos & derivados , Histidina/metabolismo , Humanos , Glomérulos Renais/patologia , Camundongos Transgênicos , Músculo Esquelético/patologia , Obesidade/complicações , Obesidade/genética , Obesidade/patologia , Fatores de Tempo
19.
Aquat Toxicol ; 222: 105471, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32199139

RESUMO

The toxicity of waterborne retene (7-isopropyl-1-methyl phenanthrene) to post-hatch embryos of rainbow trout (Oncorhynchus mykiss) was assessed at 5 and 11 °C. Survival times of retene-exposed embryos were 70 % longer at 5 °C than at 11 °C, but survival times and LC50 s did not vary when time was expressed as degree-days (thermal units), i.e., at a common stage of development. The size of survivors decreased with increasing retene concentrations, but not with temperature. Retene did not bioconcentrate to any extent (bioconcentration factors < 2) at either temperature, indicating effective biotransformation by embryos. However, concentrations of retene metabolites were slightly higher at 5 °C, suggesting slower excretion rates than at 11 °C. The relative expression of cytochrome P450 proteins (CYP1A) did not vary with temperature but increased with retene concentration, as indicated by cyp1a mRNA concentrations. The induction of CYP1A protein by retene exposure was evident in the vasculature of eye, brain, heart, kidney, liver, gill, mouth, intestine, muscle, and yolk-sac. However, immunohistochemical staining was greater at 5 than at 11 °C for all tissues except liver and muscle. Overall, temperature effects on retene toxicity disappeared when the duration of embryo development and retene exposure were expressed as thermal units (degree-days). Temperature controlled the rate of embryo development and the rate of toxicity (time to a toxic endpoint), but not the concentrations that were toxic.


Assuntos
Embrião não Mamífero/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Oncorhynchus mykiss/metabolismo , Fenantrenos/toxicidade , Temperatura , Poluentes Químicos da Água/toxicidade , Animais , Citocromo P-450 CYP1A1/biossíntese , Relação Dose-Resposta a Droga , Embrião não Mamífero/metabolismo , Indução Enzimática/efeitos dos fármacos , Dose Letal Mediana
20.
J Am Soc Nephrol ; 31(5): 983-995, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32209589

RESUMO

BACKGROUND: Expression of SerpinB2, a regulator of inflammatory processes, has been described in the context of macrophage activation and cellular senescence. Given that mechanisms for these processes interact and can shape kidney disease, it seems plausible that SerpinB2 might play a role in renal aging, injury, and repair. METHODS: We subjected SerpinB2 knockout mice to ischemia-reperfusion injury or unilateral ureteral obstruction. We performed phagocyte depletion to study SerpinB2's role beyond the effects of macrophages and transplanted bone marrow from knockout mice to wild-type mice and vice versa to dissect cell type-dependent effects. Primary tubular cells and macrophages from SerpinB2 knockout and wild-type mice were used for functional studies and transcriptional profiling. RESULTS: Cultured senescent tubular cells, kidneys of aged mice, and renal stress models exhibited upregulation of SerpinB2 expression. Functionally, lack of SerpinB2 in aged knockout mice had no effect on the magnitude of senescence markers but associated with enhanced kidney damage and fibrosis. In stress models, inflammatory cell infiltration was initially lower in knockout mice but later increased, leading to an accumulation of significantly more macrophages. SerpinB2 knockout tubular cells showed significantly reduced expression of the chemokine CCL2. Macrophages from knockout mice exhibited reduced phagocytosis and enhanced migration. Macrophage depletion and bone marrow transplantation experiments validated the functional relevance of these cell type-specific functions of SerpinB2. CONCLUSIONS: SerpinB2 influences tubule-macrophage crosstalk by supporting tubular CCL2 expression and regulating macrophage phagocytosis and migration. In mice, SerpinB2 expression seems to be needed for coordination and timely resolution of inflammation, successful repair, and kidney homeostasis during aging. Implications of SerpinB2 in human kidney disease deserve further exploration.


Assuntos
Lesão Renal Aguda/enzimologia , Envelhecimento/imunologia , Senescência Celular/imunologia , Túbulos Renais/enzimologia , Rim/enzimologia , Macrófagos/fisiologia , Inibidor 2 de Ativador de Plasminogênio/fisiologia , Traumatismo por Reperfusão/enzimologia , Obstrução Ureteral/complicações , Lesão Renal Aguda/etiologia , Lesão Renal Aguda/imunologia , Animais , Movimento Celular , Quimiocina CCL2/biossíntese , Quimiocina CCL2/genética , Técnicas de Cocultura , Indução Enzimática , Células Epiteliais/metabolismo , Fibrose , Homeostase , Rim/irrigação sanguínea , Rim/patologia , Masculino , Camundongos , Camundongos Knockout , Fagocitose , Inibidor 2 de Ativador de Plasminogênio/deficiência , Traumatismo por Reperfusão/imunologia , Transcriptoma , Obstrução Ureteral/enzimologia , Obstrução Ureteral/imunologia
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