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1.
BMC Plant Biol ; 24(1): 308, 2024 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-38644502

RESUMO

Acacia melanoxylon is well known as a valuable commercial tree species owing to its high-quality heartwood (HW) products. However, the metabolism and regulatory mechanism of heartwood during wood development remain largely unclear. In this study, both microscopic observation and content determination proved that total amount of starches decreased and phenolics and flavonoids increased gradually from sapwood (SW) to HW. We also obtained the metabolite profiles of 10 metabolites related to phenolics and flavonoids during HW formation by metabolomics. Additionally, we collected a comprehensive overview of genes associated with the biosynthesis of sugars, terpenoids, phenolics, and flavonoids using RNA-seq. A total of ninety-one genes related to HW formation were identified. The transcripts related to plant hormones, programmed cell death (PCD), and dehydration were increased in transition zone (TZ) than in SW. The results of RT-PCR showed that the relative expression level of genes and transcription factors was also high in the TZ, regardless of the horizontal or vertical direction of the trunk. Therefore, the HW formation took place in the TZ for A. melanoxylon from molecular level, and potentially connected to plant hormones, PCD, and cell dehydration. Besides, the increased expression of sugar and terpenoid biosynthesis-related genes in TZ further confirmed the close connection between terpenoid biosynthesis and carbohydrate metabolites of A. melanoxylon. Furthermore, the integrated analysis of metabolism data and RNA-seq data showed the key transcription factors (TFs) regulating flavonoids and phenolics accumulation in HW, including negative correlation TFs (WRKY, MYB) and positive correlation TFs (AP2, bZIP, CBF, PB1, and TCP). And, the genes and metabolites from phenylpropanoid and flavonoid metabolism and biosynthesis were up-regulated and largely accumulated in TZ and HW, respectively. The findings of this research provide a basis for comprehending the buildup of metabolites and the molecular regulatory processes of HW formation in A. melanoxylon.


Assuntos
Acacia , Flavonoides , Perfilação da Expressão Gênica , Madeira , Acacia/genética , Acacia/metabolismo , Flavonoides/metabolismo , Flavonoides/biossíntese , Madeira/genética , Madeira/metabolismo , Metabolômica , Regulação da Expressão Gênica de Plantas , Transcriptoma , Fenóis/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética
2.
Mediators Inflamm ; 2021: 6699499, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33510583

RESUMO

METHOD: Mice were randomly assigned to the sham, I/R, Oxy, and I/R with Oxy groups. Oxy was injected intraperitoneally 30 min before tourniquet placement. Morphological changes of the gastrocnemius muscle in these mice were assessed by hematoxylin-eosin (HE) staining and electron microscopy. Expression levels of TLR4, NF-κB, SIRT1, and PGC-1α in the skeletal muscles were detected by western blot. Blood TNF-α levels, gastrocnemius muscle contractile force, and ATP concentration were examined. RESULTS: Compared with the I/R group, Oxy pretreatment attenuated skeletal muscle damage, decreased serum TNF-α levels, and inhibited the expression levels of TLR4/NF-κB in the gastrocnemius muscle. Furthermore, Oxy treatment significantly increased serum ATP levels and the contractility of the skeletal muscles. SIRT1 and PGC-1α levels were significantly reduced in gastrocnemius muscle after I/R. Oxy pretreatment recovered these protein expression levels. CONCLUSION: Tourniquet-induced acute limb I/R results in morphological and functional impairment in skeletal muscle. Pretreatment with Oxy attenuates skeletal muscle from acute I/R injury through inhibition of TLR4/NF-κB-dependent inflammatory response and protects SIRT1/PGC-1α-dependent mitochondrial function.


Assuntos
Contração Muscular , Músculo Esquelético/metabolismo , Receptores Opioides/metabolismo , Traumatismo por Reperfusão/metabolismo , Torniquetes , Trifosfato de Adenosina/metabolismo , Animais , Inflamação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Eletrônica , Músculo Esquelético/lesões , Subunidade p50 de NF-kappa B/metabolismo , Perfusão , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Sirtuína 1/metabolismo , Receptor 4 Toll-Like/metabolismo
3.
Clin Interv Aging ; 14: 1607-1614, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564843

RESUMO

PURPOSE: The purpose of this study was to evaluate the effects of transcutaneous electrical acupoint stimulation (TEAS) on postoperative autonomic nervous system function and serum biomarkers in the elderly. PATIENTS AND METHODS: A total of 122 American Society of Anesthesiologists class II or III patients with coronary heart disease undergoing spinal surgery were randomly divided into two groups: TEAS (received TEAS at Neiguan [PC6] and Ximen [PC4] for 30 minutes before anesthesia induction until the end of surgery) and control (received electrode plate at the same acupuncture points without any electrical stimulation). Serum was isolated for the measurement of concentration of high-sensitive troponin T (hs-cTnT), CRP, and CK. Heart rate (HR) and heart rate variability (HRV) including: total power (TP), low-frequency (LF) power, high-frequency (HF) power, and LF/HF ratio were used to assess autonomic nervous system function. The primary outcome was to evaluate whether TEAS changed the postoperative serum hs-cTnT. The secondary outcomes were to observe the effects of TEAS on HRV, circulating CK and CRP after surgery. RESULTS: Hs-cTnT, CRP, and CK concentrations were significantly higher on first, third and fifth day after surgery than those before anesthesia induction in both groups. Hs-cTnT concentration was significantly lower on the first and third day after surgery in TEAS group than in control group. Compared with 1 day before surgery, TP, LF, and HF decreased significantly and HR, LF/HF increased significantly on first, third, and fifth day after surgery in control group. Compared with control group, HR was significantly lower on the first, third, and fifth day after surgery, LF/HF decreased and TP, LF, HF were significantly higher on the first day after surgery in TEAS group. CONCLUSION: TEAS at PC6 and PC4 could reduce postoperative serum hs-cTnT concentration and change HRV index to improve autonomic nervous system activity.


Assuntos
Pontos de Acupuntura , Doença das Coronárias/terapia , Assistência Perioperatória/métodos , Estimulação Elétrica Nervosa Transcutânea/métodos , Terapia por Acupuntura/métodos , Idoso , Sistema Nervoso Autônomo , Feminino , Frequência Cardíaca , Humanos , Masculino , Estudos Prospectivos
4.
Cell Death Dis ; 8(3): e2676, 2017 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-28300826

RESUMO

Inflammatory responses involving microglia and astrocytes contribute to the pathogenesis of neurodegenerative diseases (NDs). In addition, inflammation is tightly linked to iron metabolism dysregulation. However, it is not clear whether the brain inflammation-induced iron metabolism dysregulation contributes to the NDs pathogenesis. Herein, we demonstrate that the expression of the systemic iron regulatory hormone, hepcidin, is induced by lipopolysaccharide (LPS) through the IL-6/STAT3 pathway in the cortex and hippocampus. In this paradigm, activated glial cells are the source of IL-6, which was essential in the iron overload-activated apoptosis of neurons. Disrupting astrocyte hepcidin expression prevented the apoptosis of neurons, which were able to maintain levels of FPN1 adequate to avoid iron accumulation. Together, our data are consistent with a model whereby inflammation initiates an intercellular signaling cascade in which activated microglia, through IL-6 signaling, stimulate astrocytes to release hepcidin which, in turn, signals to neurons, via hepcidin, to prevent their iron release. Such a pathway is relevant to NDs in that it links inflammation, microglia and astrocytes to neuronal damage.


Assuntos
Apoptose/efeitos dos fármacos , Astrócitos/metabolismo , Hepcidinas/metabolismo , Lipopolissacarídeos/farmacologia , Neurônios/metabolismo , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Células Cultivadas , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Encefalite/metabolismo , Encefalite/patologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/patologia , Interleucina-6/metabolismo , Ferro/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microglia/efeitos dos fármacos , Microglia/metabolismo , Microglia/patologia , Neurônios/efeitos dos fármacos , Neurônios/patologia , Ratos , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos
5.
PLoS One ; 6(9): e25324, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21957487

RESUMO

Oxidative stress plays an important role in neuronal injuries caused by cerebral ischemia. It is well established that free iron increases significantly during ischemia and is responsible for oxidative damage in the brain. However, the mechanism of this ischemia-induced increase in iron is not completely understood. In this report, the middle cerebral artery occlusion (MCAO) rat model was performed and the mechanism of iron accumulation in cerebral ischemia-reperfusion was studied. The expression of L-ferritin was significantly increased in the cerebral cortex, hippocampus, and striatum on the ischemic side, whereas H-ferritin was reduced in the striatum and increased in the cerebral cortex and hippocampus. The expression level of the iron-export protein ferroportin1 (FPN1) significantly decreased, while the expression of transferrin receptor 1 (TfR1) was increased. In order to elucidate the mechanisms of FPN1 regulation, we studied the expression of the key regulator of FPN1, hepcidin. We observed that the hepcidin level was significantly elevated in the ischemic side of the brain. Knockdown hepcidin repressed the increasing of L-ferritin and decreasing of FPN1 invoked by ischemia-reperfusion. The results indicate that hepcidin is an important contributor to iron overload in cerebral ischemia. Furthermore, our results demonstrated that the levels of hypoxia-inducible factor-1α (HIF-1α) were significantly higher in the cerebral cortex, hippocampus and striatum on the ischemic side; therefore, the HIF-1α-mediated TfR1 expression may be another contributor to the iron overload in the ischemia-reperfusion brain.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Infarto da Artéria Cerebral Média/metabolismo , Ferro/metabolismo , Animais , Peptídeos Catiônicos Antimicrobianos/deficiência , Peptídeos Catiônicos Antimicrobianos/genética , Encéfalo/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Ferritinas/metabolismo , Técnicas de Silenciamento de Genes , Hepcidinas , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/genética , Interleucina-6/genética , Sobrecarga de Ferro/complicações , Sobrecarga de Ferro/genética , Sobrecarga de Ferro/metabolismo , Camundongos , Ratos , Receptores da Transferrina/metabolismo , Traumatismo por Reperfusão/complicações , Regulação para Cima
6.
Exp Neurol ; 224(2): 347-55, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20417201

RESUMO

This study investigates whether activation of p38 MAPK by the up-regulation of HSP 70 participates in the induction of brain ischemic tolerance by limb ischemic preconditioning (LIP). Western blot and immunohistochemical assays indicated that p38 MAPK activation occurred earlier than HSP 70 induction in the CA1 region of the hippocampus after LIP. P-p38 MAPK expression was up-regulated at 6h and reached its peak 12h after LIP, while HSP 70 expression was not significantly increased until 1 day and peaked 2 days after LIP. Neuropathological evaluation by thionin staining showed that quercetin (4 ml/kg, 50mg/kg, intraperitoneal injection), an inhibitor of HSP 70, blocked the protective effect of LIP against delayed neuronal death that is normally induced by lethal brain ischemic insult, indicating that HSP 70 participates in the induction of brain ischemic tolerance by LIP. Furthermore, SB 203580, an inhibitor of HSP 70, inhibited HSP 70 activation in the CA1 region of the hippocampus induced by LIP either with or without the presence of subsequent brain ischemic insult. Based on the above results, it can be concluded that activation of p38 MAPK participates in the brain ischemic tolerance induced by LIP at least partly by the up-regulation of HSP 70 expression.


Assuntos
Isquemia Encefálica/metabolismo , Proteínas de Choque Térmico HSP70/biossíntese , Membro Posterior/irrigação sanguínea , Precondicionamento Isquêmico , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Western Blotting , Isquemia Encefálica/patologia , Isquemia Encefálica/prevenção & controle , Região CA1 Hipocampal/metabolismo , Região CA1 Hipocampal/patologia , Suscetibilidade a Doenças , Ativação Enzimática , Proteínas de Choque Térmico HSP70/antagonistas & inibidores , Imidazóis/farmacologia , Imuno-Histoquímica , Masculino , Piridinas/farmacologia , Quercetina/farmacologia , Ratos , Ratos Wistar , Regulação para Cima , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores
7.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 24(7): 679-82, 2008 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-18616910

RESUMO

AIM: To establish a tumor model in HLA-A2.1 transgenic mice to examine the efficacy of MAGE-3 vaccine, a cell line coexpressing HLA-A 0201/K(b) and MAGE-3 is established. METHODS: B16-HLA-MAGE-3 melanoma was obtained by means of cotransfection of HLA-A 0201/K(b) and MAGE-3 to B16 melanoma. RT-PCR, FCM analysis and Western blot were used to detect the mRNA or protein of HLA-A 0201/K(b) or MAGE-3 expression in B16-HLA-MAGE-3. The ability of MAGE-3 antigen to be processed and presented in the B16-HLA-MAGE-3 cell line were observed by CTL activity detection and tumor challenge test. RESULTS: Transcription and protein expression of HLA-A 0201/H-2k(b) and MAGE-3 were demonstrated in B16-HLA-MAGE-3 cells. CTL activity of splenocytes in immunized mice against B16-HLA-MAGE-3 was detected and the growth of B16-HLA-MAGE-3 in immunized mice was also inhibited. CONCLUSION: MAGE-3 antigen is able to be processed and presented efficiently by B16-HLA-MAGE-3 melanoma cells and this cell can be employed to test HLA-A2 restricted epitope immunogenicity in the A2-transgenic mice.


Assuntos
Antígenos de Neoplasias/imunologia , Vacinas Anticâncer/imunologia , Antígenos HLA/imunologia , Melanoma Experimental/imunologia , Proteínas de Neoplasias/imunologia , Animais , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/metabolismo , Western Blotting , Vacinas Anticâncer/genética , Vacinas Anticâncer/metabolismo , Linhagem Celular , Antígenos HLA/genética , Antígenos HLA/metabolismo , Melanoma Experimental/metabolismo , Camundongos , Camundongos Transgênicos , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
8.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 21(2): 198-201, 2005 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-15766405

RESUMO

AIM: To prepare rabbit anti-rat P450scc antibody, and to detect the expression of rP450scc in rat brain tissue and nerve cells. METHODS: An eight-branched polypeptide, rP450scc-16 was synthesized using solid phase synthesis (Fmoc) method. A Newzealand rabbit was immunized with rP450scc-16 and the serum was separated from the whole blood 5 days after the last immunization. The titer and specificity of the antiserum were evaluated, and the expression of rP450scc in normal rat brain and primary rat astrocytes was detected by using ELISA, Western blot and immunocytochemcal staining. RESULTS: The titer of the antiserum was 1:6,400. Western blot analysis showed that the rP450scc protein in rat brain, testis and adrenal gland homogenate was recognized by the antiserum as a single band at M(r) being 50,000, which indicated a high specificity of the antiserum. Hypothalamus, cerebral cortex, and hippocampus of rat brain and the cytoplasm of cultured rat astrocytes were positively stained by the antiserum. CONCLUSION: A highly specific anti-rP450scc antibody was prepared. The antibody can be used to study the expression and distribution of rP450scc in rat brain.


Assuntos
Anticorpos/análise , Anticorpos/imunologia , Enzima de Clivagem da Cadeia Lateral do Colesterol/imunologia , Sequência de Aminoácidos , Animais , Anticorpos/química , Anticorpos/metabolismo , Especificidade de Anticorpos , Astrócitos/metabolismo , Encéfalo/citologia , Encéfalo/metabolismo , Regulação da Expressão Gênica , Imuno-Histoquímica , Masculino , Dados de Sequência Molecular , Peptídeos/análise , Peptídeos/síntese química , Peptídeos/química , Coelhos , Ratos , Ratos Sprague-Dawley
9.
Acta Pharmacol Sin ; 25(10): 1285-91, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15456529

RESUMO

AIM: To investigate the effects of morphine dependence and withdrawal on the concentrations of neurosteroids in rat brain. METHODS: A method of simultaneous quantification of neurosteroids by gas chromatography-mass spectrometry (GC-MS) had been established. RESULTS: The chronic morphine administration (ip) resulted in a marked decrease in the brain concentrations of pregnenolone (PREG), progesterone (PROG), and pregenenolone sulfate (PREGS) in rats killed 6 h after the last treatment. In contrast, there were no significant effects of morphine dependence on the brain concentrations of allopregnanolone (AP), dihydroepiandrosterone (DHEA), and dihydroepiandrosterone sulfate (DHEAS). Naloxone-induced withdrawal produced a significant increase in the concentrations of PREG, PROG, AP, DHEA, PREGS, and DHEAS as compared with the control group. CONCLUSION: Morphine dependence and withdrawal affected the concentrations of neurosteroids in rat brain, which suggests that endogenous neurosteroids in brain might be related to the development of morphine dependence and withdrawal.


Assuntos
Encéfalo/metabolismo , Dependência de Morfina/metabolismo , Pregnenolona/metabolismo , Progesterona/metabolismo , Síndrome de Abstinência a Substâncias/metabolismo , Animais , Desidroepiandrosterona/metabolismo , Sulfato de Desidroepiandrosterona/metabolismo , Masculino , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley
10.
Se Pu ; 22(1): 12-5, 2004 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-15712935

RESUMO

A simplified method has been established to simultaneously characterize different neurosteroids in rat brain by gas chromatography/mass spectrometry (GC-MS). Neurosteroids were isolated separately in a two-step procedure using ethyl acetate in the first step to extract the unconjugated steroids and chloroform/2-butanol(50/50,v/v) in the second step to extract sulfated steroids. All steroid fractions were further purified by solid phase extraction (SPE) and the sulfated steroids were solvolyzed. All steroids were derivatized with heptafluorobutyric acid anhydride and analyzed by GC-MS (electrospray ionization) using selected ion monitoring. In male rat brain, the concentrations of pregnenolone (PREG), progesterone (PROG), allo-pregnanolone (AP) and dihydroepiandrosterone (DHEA) were (8.53 +/- 1.11) ng/g, (7.01 +/- 2.60 )ng/g, (1.17 +/- 0.19 )ng/g, and (0.92 +/- 0.20) ng/g, respectively. The concentrations of pregnenolone sulfate (PREGS) and dihydroepiandrosterone sulfate (DHEAS) were (5.94 +/- 2.03) ng/g and (1.93 +/- 0.92) ng/g, respectively. Good linearity and accuracy were observed for each steroid. The procedure was suitable for measuring the concentrations of endogenous neurosteroids, simultaneously including the sulfates in rat brain.


Assuntos
Química Encefálica , Cromatografia Gasosa-Espectrometria de Massas , Esteroides/análise , Animais , Cromatografia Gasosa-Espectrometria de Massas/métodos , Masculino , Pregnanolona/análise , Progesterona/análise , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
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