RESUMEN
Cancer cachexia is a complex disorder characterized by inflammatory responses, and it is associated with poor performance status and high mortality rate of cancer patients. Carboxyamidotriazole (CAI), a noncytotoxic chemotherapy agent, shows anti-inflammatory features in the treatment of many diseases. Here, we investigated the preventive and therapeutic effects of CAI on muscle loss that occurred in mice with advanced Lewis lung carcinoma (LLC). The carcass weights of CAI-treated mice were significantly higher than that of mice in the vehicle group from Day 19 to the end of the study. The gastrocnemius and epididymal adipose tissue weights were also increased by CAI treatment. The protective mechanisms might be attributed to the following points: CAI treatment inhibited the proteolysis in muscles by decreasing expressions of muscle-specific FoxO3 transcription factor and ubiquitin E3 ligases (MuRF1 and atrogin1). Moreover, CAI restricted the NF-κB signaling, downregulated the level of TNF-α in muscle and both TNF-α and IL-6 levels in serum, directly stimulated SIRT1 activity in vitro, and increased SIRT1 content in muscle. These results indicate that CAI can alleviate muscle wasting and is a promising drug against lung cancer cachexia.
Asunto(s)
Antineoplásicos , Caquexia/tratamiento farmacológico , Atrofia Muscular/tratamiento farmacológico , FN-kappa B/antagonistas & inhibidores , Sirtuina 1/metabolismo , Triazoles , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/crecimiento & desarrollo , Animales , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Caquexia/sangre , Caquexia/etiología , Caquexia/metabolismo , Carcinoma Pulmonar de Lewis/complicaciones , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Carcinoma Pulmonar de Lewis/metabolismo , Carcinoma Pulmonar de Lewis/patología , Interleucina-6/sangre , Masculino , Ratones Endogámicos C57BL , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/crecimiento & desarrollo , Músculo Esquelético/metabolismo , Atrofia Muscular/etiología , Atrofia Muscular/metabolismo , Triazoles/farmacología , Triazoles/uso terapéutico , Carga Tumoral/efectos de los fármacos , Factor de Necrosis Tumoral alfa/sangre , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
OBJECTIVE To evaluate whether the IDO1 inhibitor 1- methyl- L- tryptophan (1- MT) combine calcium influx inhibitor carboxyamidotriazole (CAI) could further enhance the suppression of programmed death 1 (PD-1) in CD8 + T cells and investigate the curative effect of the combined use. METHODS CD8 +T cells were isolated from normal mice spleen by negative selection using magnetic cell separation. The isolated CD8 +T cells were cultured in RPMI 1640 medium containing 10% FBS and 100 U·mL- 1 IL-2 and activated by the addition of anti-CD3 and anti-CD28 (1 g·L- 1 each mabs). CD8 + T cells were pretreated for 48 h with drug and the fluo- 3 as a marker of intracellular calcium concentration was detected by flow cytometry. The calcineurin (CaN) levels were assayed with ELISA in CD8+T cells after 48 h incubation with 10 μm CAI. The nuclear translocations of NFAT and AHR were detected by immunofluorescent staining after 48 h of drug treatment. The expression of PD-1 in CD8+T cells was analyzed by flow cytometry. RESULTS Intracellular fluorescent intensity was markedly debase due to CAI treatment(P<0.01). Meanwhile, the changes of CaN content had a resembled correlation (P<0.01). Immunofluorescence experiment showed that after combination therapy the transfer of NFAT and AHR in nuclear substantially reduced. Flow cytometry revealed that after the combination caused a significant decrease in PD-1 expression in CD8+T cells. CONCLUSION CAI and 1-MT could inhibit markedly the expression of PD-1 in CD8 +T cells by inhibiting the nuclear translocation of NFAT and AHR, respectively and the combination of them has synergetic effect.
RESUMEN
OBJECTIVE: To study the effect of carboxyamidotriazole (CAI) on adjuvant arthritis (AA) in rats. METHODS: The rats were randomly divided into normal group,two vehicle groups (polyethylene glycol 400 control and normal sodium control group), CAI-treated groups (10, 20, and 40 mg/kg) and positive control dexamethasone group. Freund's completed adjuvant was used to induce AA in rats. The arthritis index (AI) was scored, and X-ray check of the hind limbs and histopathological examination were performed. The levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, and IL-6 in the inflamed paw tissues were measured. RESULTS: The administration of CAI significantly decreased the AI, restored the body weights, and ameliorated the radiological and histopathological features of joint destruction in AA rats (P<0.05, P<0.01). In addition, CAI reduced the TNF-α, IL-1ß, and IL-6 levels in the inflamed paw tissues (P<0.05, P<0.01). CONCLUSION: CAI has therapeutic effect on AA rats, which may be achieved by decreasing the pro-inflammatory cytokines at the site of inflammation.
Asunto(s)
Artritis Experimental , Animales , Adyuvante de Freund , Interleucina-1beta , Interleucina-6 , Ratas , Triazoles , Factor de Necrosis Tumoral alfaRESUMEN
OBJECTIVE: To explore the potential anti-inflammatory and analgesic activities of carboxyamidotriazole (CAI). METHODS: A variety of animal models, including the croton oil-induced ear edema, the cotton-induced granuloma, the rat adjuvant-induced arthritis, were used to evaluate anti-inflammatory effect of CAI. Vascular endothelial growth factor (VEGF)--or histamine-stimulated local vascular permeability in mouse modulated by CAI was also determined. In addition, we assessed the effect of CAI on the levels of proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-beta) at the site of inflammation and in sera. Moreover, antinociceptive effect of CAI on inflammatory pain was assessed using acetic acid-induced writhing model and the formalin test. RESULTS: CAI significantly inhibited acute and chronic phases of inflammation, reduced VEGF or histamine-induced vascular permeability, and showed marked inhibition of proinflammatory cytokines such as TNF-alpha and IL-1 beta. CAI also showed potential therapeutic effect on peripheral inflammatory pain. CONCLUSION: CAI is a promising anti-inflammatory and analgesic agent.
Asunto(s)
Analgésicos/farmacología , Antiinflamatorios/farmacología , Triazoles/farmacología , Animales , Evaluación Preclínica de Medicamentos , Femenino , Masculino , Ratones , Ratones Endogámicos ICR , Ratas , Ratas WistarRESUMEN
Carboxyamido-triazole (CAI), a voltage-independent calcium channel inhibitor, has been shown to be able to induce growth inhibition and apoptosis in cancer cells. In the present study, we demonstrate that CAI significantly inhibits proliferation of cultured MCF-7 human breast cancer cells in a dose-dependent manner with an IC(50) of approximately 26 microM. Reduced proliferation of MCF-7 cells in the presence of CAI correlated with accumulation of cells in G(2)/M phase and induction of apoptosis. A treatment of MCF-7 cells with 30 microM CAI caused a time-dependent decrease in the levels of proteins that regulate G(2)/M progression, including Cdk1, Cyclin B1, and Cdc25C. A simultaneous increase in the expression of p21 protein was observed. We also demonstrated a concurrent decrease of the mitochondrial membrane potential (DeltaPsi(m)), and down-regulation of anti-apoptotic protein Bcl-2. In conclusion, it seems reasonable to hypothesize that the antitumor effect of CAI in MCF-7 cells is based on G(2)/M cell cycle arrest and inducing apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Triazoles/farmacología , Antineoplásicos/farmacología , Western Blotting , División Celular/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , ADN de Neoplasias/biosíntesis , Relación Dosis-Respuesta a Droga , Fase G2/efectos de los fármacos , Humanos , Potenciales de la Membrana/efectos de los fármacos , Membranas Mitocondriales/efectos de los fármacos , Membranas Mitocondriales/fisiología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismoRESUMEN
AIM: To study the effects of oxyphenamone (Oxy) on activation of Ca(2+)-activated K+ channels in rabbit mesenteric vascular smooth muscle cells. METHODS: To measure the effect of Oxy on the Ca(2+)-activated K+ channel (BK (Ca) channel) activity in rabbit mesenteric vascular smooth muscle cells by using whole cell patch clamp techniques. RESULTS: Oxy reversibly increase BK (Ca) channel activity in rabbit mesenteric artery smooth muscle cells. Application of Oxy (0.1 mumol.L-1) to the perfusion solution caused significant increase in outward currents and its effect was completely abolished by washout; The outward currents K+ was inhibited by TEA (7.5 mmol.L-1); Oxy activated the BK (Ca) channel in a dose-dependent manner (0.01-10 mumol.L-1). CONCLUSION: Oxy directly increase the activity of BK (Ca) channel activity in rabbit mesenteric vascular smooth muscle cells in dose-dependent manner.
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Cardiotónicos/farmacología , Músculo Liso Vascular/citología , Miocitos del Músculo Liso/fisiología , Canales de Potasio Calcio-Activados/efectos de los fármacos , Animales , Arterias Mesentéricas/citología , Compuestos Orgánicos , Técnicas de Placa-Clamp , ConejosRESUMEN
OBJECTIVE: To determine the affinity of new opioid receptor ligands to cloned mu opioid receptors stably expressed in CHO cell. METHODS: The binding characteristics of the opioid ligand [3H] diprenorphine (3H-dip) were studied by cellular biological techniques and radioligands binding in cloned mu opioid receptors stably expressed in CHO cells in saturation binding experiments, and were followed by competition binding experiments with a variety of new synthesized opioid receptor ligands. RESULTS: The Kd and Bmax of [3H] diprenorphine bound to mu receptors were 1.06 nmol/L and 930 fmol/mg protein, respectively. Competition binding experiments revealed that ligand 3# and 12# displayed much higher affinity than DAMGO and Morphine for the cloned mu opioid receptor. However, the affinities of ligands 2#, 6#, 8# and 9# were lower than DAMGO and Morphine. CONCLUSION: The present results suggest that the new ligands 3# and 12# have higher affinity to mu opioid receptors. However, ligands 2#, 6#, 8# and 9# have lower affinity to mu opioid receptors.
Asunto(s)
Receptores Opioides mu/metabolismo , Animales , Sitios de Unión , Unión Competitiva , Células CHO/metabolismo , Clonación Molecular , Cricetinae , Diprenorfina/farmacología , Ligandos , Receptores Opioides mu/biosíntesis , Receptores Opioides mu/genéticaRESUMEN
OBJECTIVE: To investigate the dual effects by the delta opioid receptor agonists DPDPE on the delayed rectified potassium channels in NG108-15 cells. METHODS: A series of outward currents were evoked in NG108-15 cells by depolarizing voltage from -50 mV to +80 mV at holding potential of -90 mV. These currents were delayed rectified potassium currents. Relatively selected delta opioid receptor agonists DPDPE of higher and lower concentrations were used to modulate the delayed rectified K+ current in NG108-15 cells. Opioid receptor antagonist Naloxone (NAL) and relatively selected delta opioid receptor antagonist Naltrindole (NTI) were used in the present experiments for the characterization of the actions of opioid receptors. RESULTS: The relatively higher concentrations of delta opioid receptor agonist DPDPE (> or = 10(-6) mol/L) significantly increased the amplitude of the delayed rectified K+ current. On the contrary, the relatively lower concentrations of DPDPE (< or = 10(-12) mol/L) decreased the amplitude of the delayed rectified K+ current (P < 0.05). Furthermore both the increase and decrease were time-dependent. CONCLUSIONS: delta opioid receptor agonist has dual regulatory effects on the delayed rectified potassium channels in NG108-15 cells.
Asunto(s)
Encefalina D-Penicilamina (2,5)/farmacología , Naltrexona/análogos & derivados , Canales de Potasio de Rectificación Interna/efectos de los fármacos , Receptores Opioides delta/agonistas , Animales , Membrana Celular/metabolismo , Glioma/metabolismo , Glioma/patología , Células Híbridas/metabolismo , Ratones , Naloxona/farmacología , Naltrexona/farmacología , Neuroblastoma/metabolismo , Neuroblastoma/patología , Técnicas de Placa-Clamp , Canales de Potasio de Rectificación Interna/metabolismo , Ratas , Células Tumorales CultivadasRESUMEN
OBJECTIVE: To observe the role of G protein in the dual regulation of opioid receptor agonist on the delayed rectified potassium channels. METHODS: Using whole-cell patch-clamp techniques applied to NG108-15 cells, investigate the effect of opioid receptor agonist on the delayed rectified potassium channels by administration of Guanosine-5'-0'-2-thiociphosphate (GDP beta S), Pertusis Toxin (PTX), Tetroacetic acid nueleoside diphosphate kinase (NDPK) and Adenosine-3' 5' cyclic monophosphate cAMP in the pipette solution. RESULTS: (1) GDP beta S could block the changes induced by both high and low concentration of (D-Pen2.5)-enkephalin (DPDPE) (P < 0.05). (2) PTX could inhibit the excitative regulation on K+ channel by high concentration of DPDPE (P < 0.05). But CTX had no effect on K+ channel caused by DPDPE. (3) UDP could block the excitative effect of K+ channel by high concentration of NDPK, while have no changes on the inhibitory effect caused by low concentration of opioid agonists. (4) cAMP took part in the regulation in high concentration of agonist administration (P < 0.05), while no changes for low concentration of agonists. CONCLUSIONS: Dual changes were observed on delayed rectifier potassium channel by agonist treatment on NG108-15 cells. The excitative effect was Gi/o coupled in high concentration of agonist incubation, related to cAMP. While the inhibitory effect was possibly induced by G protein beta gamma subunit directly.