RESUMO
INTRODUCTION: Previous studies on the prevalence of vertical root fractures (VRFs) were based on extracted teeth, or teeth referred for apical surgery. This study examined teeth with an isolated periodontal probing depth (PD) as an indicator of VRF. The primary aim of this study was to investigate the prevalence of vertical root fracture (VRF) and non-VRF among teeth with an isolated probing depth (PD) ≥ 5 mm. The secondary aim was to assess factors associated with VRF by comparing the teeth with and without VRF in the Japanese population. METHODS: A total of 288 teeth with an isolated PD ≥ 5 mm were grouped pathologically into 8 groups comprising VRF and non-VRF conditions. A descriptive analysis for age, sex, tooth type, endodontically treated teeth (ETT) versus non-ETT, proximal contacts, PD (depth), PD (broadness and location), restoration type, and presence of a post was performed. Moreover, the associations between these factors and VRFs were investigated using the Chi-square test. RESULTS: The prevalence of VRF was 32%. Lower first molars were the most common tooth type in both VRF (31.5%) and non-VRF groups (29.7%), while premolars were exclusively frequent in VRF (30.2%) and not frequent in non-VRF (7.8%). Narrow buccolingual PD was common in VRF (78.1%) whereas wide PD was frequent in non-VRF (67.1%). ETT, narrow buccolingual PD, tooth type (premolars), restoration type (crown), and the presence of a post showed significant associations with VRF (P ≤ .001). CONCLUSIONS: VRFs may be more prevalent in ETT among the Japanese. Careful assessment is necessary to differentiate VRFs from other conditions when the lower first molars show an isolated PD ≥ 5 mm. When an endodontically treated premolar with a post shows a narrow buccolingual PD, the probability of a VRF may be greater than in other tooth types.
Assuntos
Fraturas dos Dentes , Dente não Vital , Humanos , Raiz Dentária , Fraturas dos Dentes/epidemiologia , Fraturas dos Dentes/cirurgia , Prevalência , População do Leste Asiático , Dente Pré-Molar , Dente não Vital/epidemiologia , Tomografia Computadorizada de Feixe CônicoRESUMO
The tumor suppressor p53 is involved in the DNA damage response and induces cell cycle arrest or apoptosis upon DNA damage. Drosophila p53 encodes two isoforms, p53A and p53B, that induce apoptosis in somatic cells. To investigate the roles of Drosophila p53 isoforms in female germline cells, the DNA damage response was analyzed in the adult ovary. Early oogenesis was sensitive to irradiation and lok-, p53-, and hid-dependent cell death occurred rapidly after both low- and high-dose irradiation. Both p53 isoforms were responsible for this cell death. On the other hand, delayed cell death in mid-oogenesis was induced at a low level only after high-dose irradiation in a p53-independent manner. The daily egg production, which did not change after low-dose irradiation, was severely reduced after high-dose irradiation in p53 mutant females due to the loss of germline stem cells. When the p53A or p53B isoform was expressed in the germline cells in the p53 mutant females at levels that do not affect normal oogenesis, p53A, but not p53B, restored the fertility of the irradiated female. In summary, moderate expression of p53A is critical to maintain the function of germline stem cells during normal oogenesis as well as after high-dose irradiation.
Assuntos
Apoptose/genética , Reparo do DNA , Proteínas de Drosophila/metabolismo , Drosophila/fisiologia , Oogênese/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Animais Geneticamente Modificados , Dano ao DNA/efeitos da radiação , Drosophila/efeitos da radiação , Proteínas de Drosophila/genética , Feminino , Fertilidade/genética , Fertilidade/efeitos da radiação , Masculino , Mutação , Oogênese/efeitos da radiação , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Isoformas de Proteínas/metabolismo , Espermatozoides/efeitos da radiação , Proteína Supressora de Tumor p53/genética , Irradiação Corporal TotalRESUMO
Ligation of the death receptors for TNF-α, FasL, and TRAIL triggers two common pathways, caspase-dependent intrinsic apoptosis and intracellular reactive oxygen species (ROS) generation. The apoptotic pathway is well characterized; however, a signaling linker between the death receptor and ROS production has not been clearly elucidated. Here, we found that death receptor-induced ROS generation was strongly inhibited by mitochondrial complex I and II inhibitors, but not by inhibitors of NADPH oxidase, lipoxygenase, cyclooxygenase or xanthine oxidase, indicating that ROS are mostly generated by the impairment of the mitochondrial respiratory chain. ROS generation was accompanied by caspase-8 activation, Bid cleavage, and cytochrome c release; it was blocked in FADD- and caspase-8-deficient cells, as well as by caspase-8 knockdown and inhibitor. Moreover, Bid knockdown abrogated TNF-α- or TRAIL-induced ROS generation, whereas overexpression of truncated Bid (tBid) or knockdown of cytochrome c spontaneously elevated ROS production. In addition, p53-overexpressing cells accumulated intracellular ROS via cytochrome c release mediated by the BH3-only protein Noxa induction. In a cell-free reconstitution system, caspase-8-mediated Bid cleavage and recombinant tBid induced mitochondrial cytochrome c release and ROS generation, which were blocked by Bcl-xL and antioxidant enzymes. These data suggest that anti-apoptotic Bcl-2 proteins play an important role in mitochondrial ROS generation by preventing cytochrome c release. These data provide evidence that the FADD/caspase-8/Bid/cytochrome c axis is a crucial linker between death receptors and mitochondria, where they play a role in ROS generation and apoptosis.
Assuntos
Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/genética , Caspase 8/genética , Citocromos c/genética , Mitocôndrias Hepáticas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Apoptose/genética , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/metabolismo , Caspase 8/metabolismo , Linhagem Celular Tumoral , Citocromos c/metabolismo , Proteína de Domínio de Morte Associada a Fas/genética , Proteína de Domínio de Morte Associada a Fas/metabolismo , Regulação da Expressão Gênica , Células HeLa , Humanos , Células Jurkat , Fígado/citologia , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Proteína bcl-X/genética , Proteína bcl-X/metabolismoRESUMO
AIMS: The heme oxygenase-1 (HO-1)/carbon monoxide (CO) pathway induced in astrocytes after ischemic brain injury promotes vascular endothelial growth factor (VEGF) expression to maintain and repair neurovascular function. Although HO-1-derived CO has been shown to induce hypoxia-inducible factor-1α (HIF-1α)-dependent VEGF expression, the underlying mechanism independent of HIF-1α remains to be elucidated. RESULTS: HO-1 and VEGF were coexpressed in astrocytes of ischemic mouse brain tissues. Experiments with specific siRNAs and pharmacological activators/inhibitors of various target genes demonstrated that astrocytes pre-exposed to the CO-releasing compound, CORM-2, or transfected with HO-1 increased HIF-1α-independent VEGF expression via sequential activation of the following signal cascades; Ca2+/calmodulin-dependent protein kinase kinase ß-mediated AMP-activated protein kinase (AMPK)α activation, AMPKα-induced increases in nicotinamide phosphoribosyltransferase (NAMPT) expression and cellular NAD+ level, sirtuin 1 (SIRT1)-dependent peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) stabilization and activation, and PGC-1α/estrogen-related receptor (ERR)α-mediated VEGF expression. All of these sequential events were blocked by an L-type voltage-gated Ca2+ channel inhibitor and Ca2+ chelators, but not by other Ca2+ channel inhibitors. INNOVATION: HO-1-derived CO elicits Ca2+ influx by activating L-type Ca2+ channels, which is a key player in HIF-1α-independent VEGF expression by activating the AMPKα-NAMPT-SIRT1-PGC-1α/ERRα pathway. CONCLUSION: Our results provide new mechanistic insight into the possible role for L-type Ca2+ channels in HO-1/CO-induced angiogenesis. Antioxid. Redox Signal. 27, 21-36.
Assuntos
Isquemia Encefálica/metabolismo , Canais de Cálcio Tipo L/metabolismo , Monóxido de Carbono/metabolismo , Heme Oxigenase-1/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Astrócitos/citologia , Astrócitos/patologia , Células Cultivadas , Modelos Animais de Doenças , Técnicas de Inativação de Genes , Heme Oxigenase-1/genética , Humanos , Camundongos , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Receptores de Estrogênio/metabolismo , Transdução de Sinais , Sirtuína 1/metabolismo , Receptor ERRalfa Relacionado ao EstrogênioRESUMO
AIMS: Hypoxia induces expression of various genes and microRNAs (miRs) that regulate angiogenesis and vascular function. In this study, we investigated a new functional role of new hypoxia-responsive miR-101 in angiogenesis and its underlying mechanism for regulating heme oxygenase-1 (HO-1) and vascular endothelial growth factor (VEGF) expression. RESULTS: We found that hypoxia induced miR-101, which binds to the 3'untranslated region of cullin 3 (Cul3) and stabilizes nuclear factor erythroid-derived 2-related factor 2 (Nrf2) via inhibition of the proteasomal degradation pathway. miR-101 overexpression promoted Nrf2 nuclear accumulation, which was accompanied with increases in HO-1 induction, VEGF expression, and endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) production. The elevated NO-induced S-nitrosylation of Kelch-like ECH-associated protein 1 and subsequent induction of Nrf2-dependent HO-1 lead to further elevation of VEGF production via a positive feedback loop between the Nrf2/HO-1 and VEGF/eNOS axes. Moreover, miR-101 promoted angiogenic signals and angiogenesis both in vitro and in vivo, and these events were attenuated by inhibiting the biological activity of HO-1, VEGF, or eNOS. Moreover, these effects were also observed in aortic rings from HO-1(+/-) and eNOS(-/-) mice. Local overexpression of miR-101 improved therapeutic angiogenesis and perfusion recovery in the ischemic mouse hindlimb, whereas antagomiR-101 diminished regional blood flow. INNOVATION: Hypoxia-responsive miR-101 stimulates angiogenesis by activating the HO-1/VEGF/eNOS axis via Cul3 targeting. Thus, miR-101 is a novel angiomir. CONCLUSION: Our results provide new mechanistic insights into a functional role of miR-101 as a potential therapeutic target in angiogenesis and vascular remodeling.
Assuntos
Proteínas Culina/genética , Heme Oxigenase-1/genética , MicroRNAs/genética , Neovascularização Fisiológica/genética , Fator A de Crescimento do Endotélio Vascular/genética , Animais , Hipóxia Celular/genética , Proteínas Culina/metabolismo , Fatores de Crescimento Endotelial/genética , Fatores de Crescimento Endotelial/metabolismo , Heme Oxigenase-1/metabolismo , Camundongos , MicroRNAs/biossíntese , Fator 2 Relacionado a NF-E2/genética , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The transcription factor NF-κB has an essential role in inflammation in endothelial cells. Endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) prevents vascular inflammation. However, the molecular mechanism underlying NF-κB-mediated regulation of eNOS expression has not been clearly elucidated. We here found that NF-κB-activating stimuli, such as lipopolysaccharide, tumor necrosis factor-α (TNF-α), and interleukin-1ß, suppressed eNOS mRNA and protein levels by decreasing mRNA stability, without affecting promoter activity. TNF-α-mediated suppression of eNOS expression, mRNA stability, and 3'-untranslated region (3'UTR) activity were inhibited by NF-κB inhibitors and Dicer knockdown, but not by p38 MAPK and MEK inhibitors, suggesting the involvement of NF-κB-responsive miRNAs in eNOS expression. Moreover, TNF-α increased MIR155HG expression and promoter activity as well as miR-155 biogenesis, and these increases were blocked by NF-κB inhibitors. Transfection with antagomiR-155 blocked TNF-α-mediated suppression of eNOS 3'UTR activity, eNOS mRNA and protein levels, and NO and cGMP production. These data provide evidence that NF-κB is a negative regulator of eNOS expression via upregulation of miR-155 under inflammatory conditions. These results suggest that NF-κB is a potential therapeutic target for preventing vascular inflammation and endothelial dysfunction induced by suppression of miR-155-mediated eNOS expression.
Assuntos
Óxido Nítrico Sintase Tipo III/biossíntese , Fator de Transcrição RelA/fisiologia , Regiões 3' não Traduzidas , RNA Helicases DEAD-box/genética , Técnicas de Silenciamento de Genes , Células Endoteliais da Veia Umbilical Humana , Humanos , Interleucina-1beta/farmacologia , MicroRNAs/fisiologia , Nitrilas/farmacologia , RNA Mensageiro/metabolismo , Ribonuclease III/genética , Sulfonas/farmacologia , Fator de Transcrição RelA/antagonistas & inibidores , Fator de Transcrição RelA/genética , Fator de Necrose Tumoral alfa/farmacologia , Vitanolídeos/farmacologiaRESUMO
Solid tumors supply oxygen and nutrients required for angiogenesis by producing vascular endothelial growth factor (VEGF). Thus, inhibitors of VEGF signaling abrogate tumor angiogenesis, resulting in the suppression of tumor growth and metastasis. We here investigated the effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on VEGF-induced angiogenesis. TRAIL inhibited VEGF-induced in vitro angiogenesis of human umbilical vein endothelial cells (HUVECs) and in vivo neovascularization in chicken embryos and mice. TRAIL blocked VEGF-induced angiogenic signaling by inhibiting ERK, Src, FAK, paxillin, Akt, and eNOS. Further, TRAIL blocked intracellular Ca(2+) elevation and actin reorganization in HUVECs stimulated with VEGF, without inhibiting VEGF receptor-2 tyrosine phosphorylation. TRAIL increased caspase-8 activity, without inducing caspase-9/-3 activation and apoptosis. Moreover, TRAIL resulted in cleavage of FAK into FAK-related non-kinase-like fragments in VEGF-stimulated HUVECs, which was blocked by a caspase-8 inhibitor and cellular caspase-8-like inhibitory protein. Biochemical and pharmacological inhibition of caspase-8 and FAK blocked the inhibitory effects of TRAIL on VEGF-stimulated anti-angiogenic signaling and events. In addition, caspase-8 knockdown also suppressed VEGF-mediated signaling and angiogenesis, suggesting that procaspase-8 plays a role of a non-apoptotic modulator in VEGF-induced angiogenic signaling. These results suggest that TRAIL inhibits VEGF-induced angiogenesis by increasing caspase-8 activity and subsequently decreasing non-apoptotic signaling functions of procaspase-8, without inducing caspase-3 activation and endothelial cell cytotoxicity. These data indicate that caspase-8 may be used as an anti-angiogenic drug for solid tumors resistant to TRAIL and anti-tumor drugs.
Assuntos
Caspase 8/metabolismo , Neovascularização Fisiológica/fisiologia , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Caspase 8/genética , Embrião de Galinha , Células HeLa , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ligante Indutor de Apoptose Relacionado a TNF/genética , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The fruit of Rubus coreanus has been used as a traditional herbal medicine for alleviation of inflammatory and vascular diseases in Asian countries. AIM OF THE STUDY: The anti-atherogenic effect of unripe Rubus coreanus fruit extract (URFE) and its underlying mechanism were analyzed in mice fed a high-fat diet (HFD) and in cell culture system. MATERIALS AND METHODS: Mouse was freely given HFD alone or supplemented with URFE for 14 weeks, followed by analysis of atherosclerotic lesions and serum lipid levels. For in vitro assay, macrophages were pretreated with URFE, followed by stimulation with lipopolysaccharide (LPS). Expression levels of inflammatory genes (TNF-α, IL-1ß, and iNOS) and phase II genes (heme oxygenase-1, glutamate cysteine lygase, and peroxiredoxine-1) as well as intracellular reactive oxygen species (ROS) level and NF-κB activation pathway were analyzed in cultured macrophages as well as mouse sera and aortic tissues. RESULTS: URFE supplementation reduced HFD-induced atherosclerotic lesion formation which was correlated with decreased levels of lipids, lipid peroxides, and inflammatory mediators (TNF-α, IL-1ß, and nitric oxide) in sera as well as suppression of inflammatory gene in aortic tissues. In addition, pre-treatment of macrophages with URFE also suppressed LPS-induced NF-κB activation, ROS production, and inflammatory and phase II gene expressions. Inhibition of phase II enzyme and protein activities attenuated the suppressive effects URFE on ROS production, NF-κB activation, and inflammatory gene expression. CONCLUSION: These results suggest that URFE attenuates atherosclerosis by improving blood lipid profile and inhibiting NF-κB activation via phase II antioxidant gene expression.
Assuntos
Aterosclerose/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Rosaceae , Animais , Aorta/metabolismo , Aterosclerose/genética , Aterosclerose/metabolismo , Linhagem Celular , Colesterol/sangue , Frutas , Regulação da Expressão Gênica/efeitos dos fármacos , Glutamato-Cisteína Ligase/genética , Heme Oxigenase-1/genética , Proteínas de Homeodomínio/genética , Interleucina-1beta/metabolismo , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Keap1 is a cytoplasmic repressor of the transcription factor Nrf2, and its degradation induces Nrf2 activation, leading to upregulation of antioxidant phase II genes. We investigated the roles of phase II genes in vascular inflammation and septic injury using Keap1 siRNA and elucidated its underlying mechanism. Selective knockdown of Keap1 with siRNA promoted Nrf2-dependent expression of phase II genes in endothelial cells, such as heme oxygenase-1 (HO-1), glutamate-cysteine ligase (GCL), and peroxiredoxin-1 (Prx1), resulting in the elevation of cellular glutathione levels and suppression of tumor necrosis factor (TNF)-α-induced intracellular H(2)O(2) accumulation. Keap1 knockdown inhibited TNF-α-induced expression of intracellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) by suppressing NF-κB activation via inhibition of its upstream modulators, Akt, NIK, and IKK, resulting in the elevation of monocyte adhesion to endothelial cells. Importantly, these events were reversed by HO-1 and GCL inhibitors and Prx1-specific siRNA. Keap1 knockdown also inhibited endotoxin-induced expression of inducible nitric oxide synthase (iNOS) and TNF-α by upregulating HO-1, GCL, and Prx1 expression in macrophages. Moreover, in vivo Keap1 knockdown increased the expression of phase II genes and suppressed the expression of ICAM-1, VCAM-1, iNOS, and TNF-α in an endotoxemic mouse model, resulting in significant protection against liver and lung injuries and lethality. Our results indicate that Keap1 knockdown prevents NF-κB-mediated vascular inflammation and endotoxic shock by suppressing NF-κB-mediated inflammatory gene expression via upregulation of Nrf2-mediated antioxidant genes. Thus, siRNA targeting Keap1 may provide a new therapeutic approach for inflammation-associated vascular diseases and sepsis.
Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/genética , Desintoxicação Metabólica Fase II/genética , Fator 2 Relacionado a NF-E2/metabolismo , RNA Interferente Pequeno/genética , Vasculite/metabolismo , Animais , Elementos de Resposta Antioxidante , Adesão Celular , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Células Cultivadas , Citoproteção , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Glutamato-Cisteína Ligase/genética , Glutamato-Cisteína Ligase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Células Endoteliais da Veia Umbilical Humana/imunologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch , Lipopolissacarídeos/farmacologia , Fígado/imunologia , Fígado/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Peroxirredoxinas/genética , Peroxirredoxinas/metabolismo , Interferência de RNA , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfa/fisiologia , Vasculite/imunologia , Vasculite/patologiaRESUMO
Androgen receptor (AR) signaling plays a critical role in the development and progression of prostate cancer. It has been reported previously that peroxiredoxin-1 (Prx1), a member of a novel family of peroxidases, interacts physically with AR to enhance AR transactivation of target genes. In the present study, we evaluated the biological significance of Prx1 in modulating dihydrotestosterone (DHT)-stimulated growth and AR target gene expression of prostate cancer cells. We also investigated the mechanism by which Prx1 might potentiate AR signaling. The contribution of Prx1 was assessed mainly by using the approach of stable Prx1 knockdown. The major observations are as follows: (a) A low level of Prx1 desensitizes cells to growth stimulation and AR target gene induction by DHT, such that exposure to a higher level of DHT is required to reach the same magnitude of response when Prx1 is depressed; (b) Prx1 increases the affinity of AR to DHT and decreases the rate of DHT dissociation from the occupied receptor; (c) Prx1 enhances the NH2 terminus and COOH terminus interaction of AR; a stronger N-C interaction is consistent with a more robust AR activation signal by keeping DHT tight in the ligand-binding pocket; (d) the stimulatory effects of Prx1 on AR ligand binding affinity and AR N-C interaction are manifested regardless of a wild-type or mutant AR. The above findings led us to believe that Prx1 may be a therapeutic target in blocking the transition of prostate cancer from an androgen-dependent to an androgen-refractory phenotype.
Assuntos
Di-Hidrotestosterona/farmacologia , Peroxirredoxinas/metabolismo , Neoplasias da Próstata/metabolismo , Receptores Androgênicos/metabolismo , Processos de Crescimento Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ciclina D1/metabolismo , Di-Hidrotestosterona/metabolismo , Dimerização , Técnicas de Silenciamento de Genes , Humanos , Masculino , Peroxirredoxinas/genética , Antígeno Prostático Específico/metabolismo , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/patologia , Ligação Proteica , Interferência de RNA , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Calicreínas Teciduais/metabolismoRESUMO
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a member of the tumor necrosis factor gene family, is considered as one of the most promising cancer therapeutic agents due to its ability to selectively induce tumor cell apoptosis. In this study, we investigated whether the Na(+)/H(+) exchanger inhibitor, amiloride, promotes TRAIL-induced apoptotic death both in sensitive and resistant tumor cells, HeLa and LNCaP cells, respectively, and its underlying molecular mechanism. Amiloride enhanced TRAIL-induced apoptosis and activation of caspase-3 and -8 in both cells. This compound increased TRAIL-induced mitochondrial cytochrome c release and poly(ADP-ribose) polymerase cleavage. Moreover, amiloride-induced intracellular acidification, and inhibited the phosphorylated activation of the serine/threonine kinase Akt, which is known to promote cell survival, in both tumor cells. These data suggest that amiloride sensitizes both tumor cells to TRAIL-induced apoptosis by promoting Akt dephosphorylation and caspase-8 activation via the intracellular acidification and that Na(+)/H(+) exchanger inhibitors may play an important role in the anti-cancer activity of TRAIL, especially, in TRAIL-resistant tumors with highly active and expressed Akt.
Assuntos
Amilorida/farmacologia , Apoptose/efeitos dos fármacos , Líquido Intracelular/química , Glicoproteínas de Membrana/farmacologia , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/patologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Reguladoras de Apoptose , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Ativação Enzimática/efeitos dos fármacos , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Líquido Intracelular/efeitos dos fármacos , Masculino , Neoplasias da Próstata/química , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt , Ligante Indutor de Apoptose Relacionado a TNFRESUMO
OBJECTIVE: The purpose of this study was to investigate the age-related morphologic changes of the lateral retropharyngeal lymph nodes using MR imaging. STUDY DESIGN: The maximal axial diameter of the nodes was measured on the MR image in 120 healthy subjects (younger group 6-19 years (n = 40), middle group 20-38 years (n = 48), older group 42-74 years (n = 32)). Between-group differences in the diameter of the nodes were analyzed. RESULTS: Mean values of the maximal axial diameter of the nodes were 9.0 +/- 1.6 mm, 6.6 +/- 1.7 mm, and 5.3 +/- 1.6 mm, corresponding to younger group, middle group, and older group, respectively ( P <.01). CONCLUSION: The information obtained in the current study is useful for the differential diagnosis of normal and abnormal lateral retropharyngeal lymph nodes according to age.
Assuntos
Linfonodos/anatomia & histologia , Linfonodos/crescimento & desenvolvimento , Desenvolvimento Maxilofacial , Adolescente , Adulto , Fatores Etários , Idoso , Análise de Variância , Criança , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Faringe/anatomia & histologia , Estatísticas não ParamétricasRESUMO
OBJECTIVE: This study aimed to evaluate the usefulness of suitable conventional spin-echo (CSE) and fast spin-echo (FSE) T2-weighted imaging parameters for the assessment of joint effusion in a phantom study and in a comparative study of CSE and FSE using clinical cases. STUDY DESIGN: In the phantom study, the signal ratios of water and oil signal fields were determined and studied comparatively. The shape and size of signals were evaluated separately. In the study of joint effusion images, 318 joints were evaluated. CSE T2-weighted imaging and FSE T2-weighted imaging were carried out, and a comparative assessment was performed. RESULTS: In both CSE and FSE imaging, the ratios of mean MRI signal values showed divergence as TR/TE values increased. The evaluation of joint effusion with FSE TR/TE 8000/120 msec was significantly better than that in all other groups (P <.01). CONCLUSION: The use of FSE requires investigation of TR/TE values. When a 0.5 T static field strength MRI apparatus is employed, TR/TE 8000/120 msec is recommended.
Assuntos
Aumento da Imagem/métodos , Imageamento por Ressonância Magnética/métodos , Líquido Sinovial , Articulação Temporomandibular/patologia , Adolescente , Adulto , Idoso , Criança , Feminino , Humanos , Masculino , Côndilo Mandibular/patologia , Pessoa de Meia-Idade , Variações Dependentes do Observador , Imagens de Fantasmas , Disco da Articulação Temporomandibular/patologia , Transtornos da Articulação Temporomandibular/patologiaRESUMO
Complestatin, a bicyclo hexapeptide from Streptomyces, was isolated as a possible regulator of neuronal cell death. In this study, we report an anti-apoptotic activity of complestatin and its underlying molecular mechanism. Complestatin blocked TRAIL (TNF-related apoptosis-inducing ligand)-induced apoptosis and activation of caspase-3 and -8 at micromolar concentration levels without inhibiting the catalytic activities of these caspases. Complestatin potently induced a rapid and sustained AKT/PKB activation and Bad phosphorylation, resulting in inhibition of mitochondrial cytochrome c release. These anti-apoptotic activities of complestatin were significantly abrogated in cells expressing dominant negative AKT/PKB. Taken together, our results suggest that complestatin prevents apoptotic cell death via AKT/PKB-dependent inhibition of the mitochondrial apoptosis signal pathway. The novel property of complestatin may be valuable for developing new pharmaceutical means that will control unwanted cell death.
Assuntos
Apoptose/efeitos dos fármacos , Inibidores de Caspase , Clorofenóis/farmacologia , Mitocôndrias/enzimologia , Peptídeos Cíclicos/farmacologia , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Reguladoras de Apoptose , Proteína Agonista de Morte Celular de Domínio Interatuante com BH3 , Proteínas de Transporte/antagonistas & inibidores , Caspases/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Citocromos c/antagonistas & inibidores , Eletroforese em Gel de Poliacrilamida , Ativação Enzimática/efeitos dos fármacos , Células HeLa , Humanos , Immunoblotting , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/metabolismo , Mitocôndrias/efeitos dos fármacos , Fosforilação , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt , Ligante Indutor de Apoptose Relacionado a TNF , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismoRESUMO
OBJECTIVE: We sought to determine the suitable magnetic resonance imaging conditions for the short inversion time inversion recovery (STIR) sequence through the use of phantoms; to describe the signal characteristics of normal structures in the mandible; and to evaluate the usefulness of STIR images in enabling the identification of mandibular osteomyelitis on conventional T1- and T2-weighted spin-echo images. STUDY DESIGN: Suitable mandibular STIR imaging conditions were determined by varying inversion time and repetition time in each sequence. STIR magnetic resonance images of 162 healthy subjects and STIR and T1- and T2-weighted spin-echo images of 21 subjects with mandibular osteomyelitis were evaluated. RESULTS: In STIR imaging, the signal of oil was suppressed at an inversion time equaling 100 milliseconds and a repetition time equaling 1500 to 3000 milliseconds. In healthy subjects, the mandibular marrow was revealed to have high signal intensities (100%) and cortical bone had no signal intensities (100%) on STIR images. In surrounding soft tissue in these healthy subjects, the submandibular glands were shown to have high signal intensities (100%); the parotid glands had intermediate to high signal intensities (100%); the sublingual glands had high (88.9%) and intermediate to high (11.1%) signal intensities; lymph nodes had high signal intensities (100%); and the masseter muscles had intermediate signal intensities (100%) on STIR images. The lesions in bone marrow had low (75%) and low to intermediate (25%) signal intensities on T1-weighted images and high (54%), intermediate to high (29%), and intermediate (17%) signal intensities on T2-weighted images. On STIR images, the signal intensities resulted in high (75%), intermediate to high (21%), and intermediate (4%) signal intensities. CONCLUSIONS: STIR imaging is highly effective for the evaluation of bone marrow and surrounding soft tissue in terms of the detection of osteomyelitis in the mandible and the identification of inflammation spreading to soft tissue.
Assuntos
Imageamento por Ressonância Magnética/métodos , Mandíbula/anatomia & histologia , Mandíbula/patologia , Doenças Mandibulares/diagnóstico , Osteomielite/diagnóstico , Adulto , Idoso , Medula Óssea/patologia , Feminino , Humanos , Aumento da Imagem , Masculino , Pessoa de Meia-Idade , Imagens de FantasmasRESUMO
Astaxanthin, a carotenoid without vitamin A activity, has shown anti-oxidant and anti-inflammatory activities; however, its molecular action and mechanism have not been elucidated. We examined in vitro and in vivo regulatory function of astaxanthin on production of nitric oxide (NO) and prostaglandin E2 (PGE2) as well as expression of inducible NO synthase (iNOS), cyclooxygenase-2, tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Astaxanthin inhibited the expression or formation production of these proinflammatory mediators and cytokines in both lipopolysaccharide (LPS)-stimulated RAW264.7 cells and primary macrophages. Astaxanthin also suppressed the serum levels of NO, PGE2, TNF-alpha, and IL-1beta in LPS-administrated mice, and inhibited NF-kappaB activation as well as iNOS promoter activity in RAW264.7 cells stimulated with LPS. This compound directly inhibited the intracellular accumulation of reactive oxygen species in LPS-stimulated RAW264.7 cells as well as H2O2-induced NF-kappaB activation and iNOS expression. Moreover, astaxanthin blocked nuclear translocation of NF-kappaB p65 subunit and I(kappa)B(alpha) degradation, which correlated with its inhibitory effect on I(kappa)B kinase (IKK) activity. These results suggest that astaxanthin, probably due to its antioxidant activity, inhibits the production of inflammatory mediators by blocking NF-kappaB activation and as a consequent suppression of IKK activity and I(kappa)B-alpha degradation.