RESUMEN

Lung transplantation stands as a vital treatment for severe lung diseases, primarily sourcing organs from donors with brain death (BD). This research delved into the potential anti-inflammatory effects of thalidomide in rats with BD-induced lung complications. In this study twenty-four Wistar rats were divided into three groups: the control (CTR), brain death (BD) and brain death + thalidomide (TLD) groups. Post specific procedures, a 360 min monitoring period ensued. Comprehensive analyses of blood and heart-lung samples were conducted. Elevated IL-6 levels characterized both BD and TLD groups relative to the CTR (p = 0.0067 and p = 0.0137). Furthermore, TNF-α levels were notably higher in the BD group than both CTR and TLD (p = 0.0152 and p = 0.0495). Additionally, IL-1ß concentrations were significantly pronounced in both BD and TLD compared to CTR, with the BD group surpassing TLD (p = 0.0256). Immunohistochemical assessments revealed augmented NF-ĸB expression in the BD group in comparison to both CTR and TLD (p = 0.0006 and p = 0.0005). With this study we can conclude that BD induced acute pulmonary inflammation, whereas thalidomide manifested a notable capability in diminishing key inflammatory markers, indicating its prospective therapeutic significance in lung transplantation scenarios.

Asunto(s)

Muerte Encefálica , Talidomida , Ratas , Animales , Talidomida/farmacología , Ratas Wistar , Muerte Encefálica/metabolismo , Pulmón/metabolismo , Antiinflamatorios/farmacología

RESUMEN

PURPOSE: To understand the effect of Nitazoxanide (NTZ), Rapamycin, Thalidomide, alone and in combination with BCG on bladder cancer (BC) histopathology and programmed death-ligand 1 (PD-L1) and anti-cytotoxic T lymphocyte antigen 4 (CTLA4) expression. METHODS: Female Fisher-344 rats underwent intravesical N-methyl-N-nitrosourea (MNU) followed by weekly intravesical treatment with saline (controls, n = 10), BCG (n = 10), NTZ (n = 8), BCG plus NTZ (n = 8), Rapamycin (n = 10) BCG plus Rapamycin (n = 10), Thalidomide (n = 10), and BCG plus Thalidomide (n = 10), and euthanized after 8 weeks and their bladders were investigated for BC and PD-L1 and CTLA4 expression. RESULTS: Rapamicyn alone and in combination with BCG had the lowest number of bladder neoplasias in the histopathology exam (1/10). Neoplastic lesions were found in 4/10 BCG recipients, 5/10 Thalidomide recipients, 4/10 Thalidomide plus BCG recipients, 5/8 NTZ and 3/8 NTZ plus BCG recipients. Adding NTZ to BCG increased the expression of PD-L1 and adding Rapamycin or Thalidomide decreased PD-L1 and CTLA4 expression compared to BCG alone. Rapamycin alone significantly increased CTLA4 and slightly increased PD-L1 expression but its combination with BCG significantly decreased both markers. Thalidomide had a similar effect; however, it was only slightly different from the control and BCG alone groups. CONCLUSION: Intravesical BCG combination treatment seems to effectively prevent BC development in an immunecompetent clinically relevant animal model, introducing Thalidomide, Nitazoxanide, and specially Rapamycin as candidates in the intravesical immunotherapy advancement. Our study contributes in understanding the mechanism of cancer immunotherapy.

Asunto(s)

Talidomida , Neoplasias de la Vejiga Urinaria , Ratas , Femenino , Animales , Talidomida/farmacología , Talidomida/uso terapéutico , Vacuna BCG/uso terapéutico , Antígeno B7-H1 , Antígeno CTLA-4 , Sirolimus/farmacología , Sirolimus/uso terapéutico , Neoplasias de la Vejiga Urinaria/patología , Administración Intravesical , Adyuvantes Inmunológicos/uso terapéutico

RESUMEN

Several molecular mechanisms of thalidomide embryopathy (TE) have been investigated, from anti-angiogenesis to oxidative stress to cereblon binding. Recently, it was discovered that thalidomide and its analogs, named immunomodulatory drugs (IMiDs), induced the degradation of C2H2 transcription factors (TFs). This mechanism might impact the strict transcriptional regulation of the developing embryo. Hence, this study aims to evaluate the TFs altered by IMiDs, prioritizing the ones associated with embryogenesis through transcriptome and systems biology-allied analyses. This study comprises only the experimental data accessed through bioinformatics databases. First, proteins and genes reported in the literature as altered/affected by the IMiDs were annotated. A protein systems biology network was evaluated. TFs beta-catenin (CTNNB1) and SP1 play more central roles: beta-catenin is an essential protein in the network, while SP1 is a putative C2H2 candidate for IMiD-induced degradation. Separately, the differential expressions of the annotated genes were analyzed through 23 publicly available transcriptomes, presenting 8624 differentially expressed genes (2947 in two or more datasets). Seventeen C2H2 TFs were identified as related to embryonic development but not studied for IMiD exposure; these TFs are potential IMiDs degradation neosubstrates. This is the first study to suggest an integration of IMiD molecular mechanisms through C2H2 TF degradation.

Asunto(s)

Mieloma Múltiple , Talidomida , Humanos , Talidomida/farmacología , Agentes Inmunomoduladores , beta Catenina/genética , beta Catenina/metabolismo , Factores de Transcripción/metabolismo , Biología de Sistemas , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Factores Inmunológicos/farmacología , Factores Inmunológicos/química , Ubiquitina-Proteína Ligasas/metabolismo , Mieloma Múltiple/metabolismo

RESUMEN

Thalidomide (TAL) has shown potential therapeutic effects in neurological diseases like epilepsy. Both clinical and preclinical studies show that TAL may act as an antiepileptic drug and as a possible treatment against disease development. However, the evidence for these effects is limited. Therefore, the antiepileptogenic and anti-inflammatory effects of TAL were evaluated herein. Sprague Dawley male rats were randomly allocated to one of five groups (n = 18 per group): control (C); status epilepticus (SE); SE-TAL (25 mg/kg); SE-TAL (50 mg/kg); and SE-topiramate (TOP; 60mg/kg). The lithium-pilocarpine model was used, and one day after SE induction the rats received pharmacological treatment for one week. The brain was obtained, and the hippocampus was micro-dissected 8, 18, and 28 days after SE. TNF-α, IL-6, and IL-1ß concentrations were quantified. TOP and TAL (50 mg/kg) increased the latency to the first of many spontaneous recurrent seizures (SRS) and decreased SRS frequency, as well as decreasing TNF-α and IL-1ß concentrations in the hippocampus. In conclusion, the results showed that both TAL (50 mg/kg) and TOP have anti-ictogenic and antiepileptogenic effects, possibly by decreasing neuroinflammation.

Asunto(s)

Encefalitis , Epilepsia del Lóbulo Temporal , Estado Epiléptico , Ratas , Masculino , Animales , Pilocarpina/toxicidad , Litio/farmacología , Litio/uso terapéutico , Talidomida/farmacología , Talidomida/uso terapéutico , Ratas Sprague-Dawley , Factor de Necrosis Tumoral alfa/metabolismo , Epilepsia del Lóbulo Temporal/inducido químicamente , Epilepsia del Lóbulo Temporal/tratamiento farmacológico , Convulsiones/inducido químicamente , Convulsiones/tratamiento farmacológico , Estado Epiléptico/inducido químicamente , Estado Epiléptico/tratamiento farmacológico , Hipocampo/metabolismo , Modelos Animales de Enfermedad

RESUMEN

Hepatocellular carcinoma (HCC) is the most common form of liver cancer. The number of cases is increasing and the trend for the next few years is not encouraging. HCC is usually detected in the advanced stages of the disease, and pharmacological therapies are not entirely effective. For this reason, it is necessary to search for new therapeutic options. The objective of this work was to evaluate the effect of the drugs isotretinoin and thalidomide on c-MYC expression and cancer-related proteins in an HCC cellular model. The expression of c-MYC was measured using RT-qPCR and western blot assays. In addition, luciferase activity assays were performed for the c-MYC promoters P1 and P2 using recombinant plasmids. Dose-response-time analyses were performed for isotretinoin or thalidomide in cells transfected with the c-MYC promoters. Finally, a proteome profile analysis of cells exposed to these two drugs was performed and the results were validated by western blot. We demonstrated that in HepG2 cells, isotretinoin and thalidomide reduced c-MYC mRNA expression levels, but this decrease in expression was linked to the regulation of P1 and P1-P2 c-MYC promoter activity in isotretinoin only. Thalidomide did not exert any effect on c-MYC promoters. Also, isotretinoin and thalidomide were capable of inducing and repressing proteins associated with cancer. In conclusion, isotretinoin and thalidomide down-regulate c-MYC mRNA expression and this is partially due to P1 or P2 promoter activity, suggesting that these drugs could be promising options for modulating the expression of oncogenes and tumor suppressor genes in HCC.

Asunto(s)

Carcinoma Hepatocelular/metabolismo , Isotretinoína/farmacología , Neoplasias Hepáticas/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Talidomida/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Carcinoma Hepatocelular/genética , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamiento farmacológico , Neoplasias Hepáticas/genética , Regiones Promotoras Genéticas , Proteómica/métodos

RESUMEN

PURPOSE: Chemotherapy for advanced pancreatic cancer has limited efficacy due to the difficultly of treating established tumours and the evolution of tumour resistance. Chemotherapies for pancreatic cancer are typically studied for their cytotoxic properties rather than for their ability to increase the immunogenicity of pancreatic tumour cells. In this study Gemcitabine in combination with immune modulatory chemotherapies Oxaliplatin, zoledronic acid and pomalidomide was studied to determine how combination therapy alters the immunogenicity of pancreatic tumour cell lines and subsequent T-cell responses. METHODS: Pancreatic tumour cell lines were stimulated with the chemotherapeutic agents and markers of immune recognition were assessed. The effect of chemotherapeutic agents on DC function was measured using uptake of CFSE-stained PANC-1 cells, changes in markers of maturation and their ability to activate CD8+ T-cells. The effect of chemotherapeutic agents on T-cell priming prior to activation using anti-CD3 and anti-CD28 antibodies was determined by measuring IFN-γ expression and Annexin V staining using flow cytometry. RESULTS: These agents demonstrate both additive and inhibitory properties on a range of markers of immunogenicity. Gemcitabine was notable for its ability to induce the upregulation of human leukocyte antigen and checkpoints on pancreatic tumour cell lines whilst inhibiting T-cell activation. Pomalidomide demonstrated immune modulatory properties on dendritic cells and T-cells, even in the presence of gemcitabine. DISCUSSION: These data highlight the complex interactions of different agents in the modulation of tumour immunogenicity and immune cell activation and emphasise the complexity in rationally designing chemo immunogenic combinations for use with immunotherapy.

Asunto(s)

Antimetabolitos Antineoplásicos/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Desoxicitidina/análogos & derivados , Inmunomodulación/efectos de los fármacos , Neoplasias Pancreáticas/inmunología , Anexina A5/metabolismo , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Línea Celular Tumoral , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Desoxicitidina/farmacología , Interacciones Farmacológicas/inmunología , Antígenos de Histocompatibilidad Clase I/efectos de los fármacos , Antígenos de Histocompatibilidad Clase I/metabolismo , Humanos , Inmunomodulación/inmunología , Interferón gamma/metabolismo , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Oxaliplatino/farmacología , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/patología , Talidomida/análogos & derivados , Talidomida/farmacología , Ácido Zoledrónico/farmacología , Gemcitabina

RESUMEN

Reversing the developmental switch from fetal hemoglobin (HbF, α2γ2) to adult hemoglobin (HbA, α2ß2) is an important therapeutic approach in sickle cell disease (SCD) and ß-thalassemia. In healthy individuals, SCD patients, and patients treated with pharmacologic HbF inducers, HbF is present only in a subset of red blood cells known as F cells. Despite more than 50 years of observations, the cause for this heterocellular HbF expression pattern, even among genetically identical cells, remains unknown. Adult F cells might represent a reversion of a given cell to a fetal-like epigenetic and transcriptional state. Alternatively, isolated transcriptional or posttranscriptional events at the γ-globin genes might underlie heterocellularity. Here, we set out to understand the heterogeneity of HbF activation by developing techniques to purify and profile differentiation stage-matched late erythroblast F cells and non-F cells (A cells) from the human HUDEP2 erythroid cell line and primary human erythroid cultures. Transcriptional and proteomic profiling of these cells demonstrated very few differences between F and A cells at the RNA level either under baseline conditions or after treatment with HbF inducers hydroxyurea or pomalidomide. Surprisingly, we did not find differences in expression of any known HbF regulators, including BCL11A or LRF, that would account for HbF activation. Our analysis shows that F erythroblasts are not significantly different from non-HbF-expressing cells and that the primary differences likely occur at the transcriptional level at the ß-globin locus.

Asunto(s)

Eritroblastos/metabolismo , Hemoglobina Fetal/biosíntesis , Hemoglobina A/metabolismo , Adulto , Anemia de Células Falciformes/sangre , Anemia de Células Falciformes/genética , Línea Celular , Separación Celular/métodos , Células Cultivadas , Eritroblastos/clasificación , Eritroblastos/efectos de los fármacos , Células Eritroides/clasificación , Células Eritroides/metabolismo , Hemoglobina Fetal/genética , Perfilación de la Expresión Génica , Hemoglobina A/genética , Humanos , Hidroxiurea/farmacología , Proteómica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Talidomida/análogos & derivados , Talidomida/farmacología

RESUMEN

OBJECTIVE: To analyze the effect of thalidomide on the progression of endometriotic lesions experimentally induced in rats and to characterize the pattern of cell proliferation by immunohistochemical Proliferating Cell Nuclear Antigen (PCNA) labeling of eutopic and ectopic endometrium. METHODS: Fifteen female Wistar rats underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium and fixation of a tissue segment to the pelvic peritoneum. Four weeks after, the animals were divided into 3 groups: control (I), 10mg/kg/day (II) and 1mg/kg/day (III) intraperitoneal thalidomide for 10 days. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma analysis. Eutopic and ectopic endometrial tissue was submitted to immunohistochemistry for analysis of cell proliferation by PCNA labeling and the cell proliferation index (CPI) was calculated as the number of labeled cells per 1,000 cells. RESULTS: Group I showed a mean CPI of 0.248 ± 0.0513 in the gland and of 0.178 ± 0.046 in the stroma. In contrast, Groups II and III showed a significantly lower CPI, that is, 0.088 ± 0.009 and 0.080 ± 0.021 for the gland (p < 0.001) and 0.0945 ± 0.0066 and 0.075 ± 0.018 for the stroma (p < 0.001), respectively. Also, the mean lesion area of Group I was 69.2 mm2, a significantly higher value compared with Group II (49.4 mm2, p = 0.023) and Group III (48.6 mm2, p = 0.006). No significant difference was observed between Groups II and III. CONCLUSION: Thalidomide proved to be effective in reducing the lesion area and CPI of the experimental endometriosis implants both at the dose of 1 mg/kg/day and at the dose of 10 mg/kg/day.

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OBJETIVO: Analisar o efeito da talidomida na progressão de lesões endometrióticas induzidas experimentalmente em ratas e caracterizar o padrão de proliferação celular pela marcação imunohistoquímica de Antígeno Nuclear de Célula Proliferativa (PCNA) no endométrio eutópico e ectópico. MéTODOS: Quinze ratas Wistar foram submetidas a laparotomia para indução de endometriose por ressecção de um corno uterino, isolamento do endométrio e fixação de um segmento do tecido ao peritônio pélvico. Após quatro semanas, os animais foram divididos em 3 grupos: controle (I), 10 mg/kg/dia (II) e 1 mg/kg/dia (III) de talidomida intraperitoneal por um período de 10 dias. As lesões foram resseccionadas juntamente com o corno uterino oposto para análise da glândula endometrial e do estroma. O tecido endometrial eutópico e ectópico foi submetido à imunohistoquímica para análise da proliferação celular por marcação com PCNA e o índice de proliferação celular (CPI) foi calculado como o número de células marcadas por 1.000 células. RESULTADOS: O grupo I apresentou média de CPI de 0,248 ± 0,0513 na glândula e de 0,178 ± 0,046 no estroma. Em contraste, os grupos II e III apresentaram CPI significativamente menor, isto é, 0,088 ± 0,009 e 0,080 ± 0,021 para a glândula (p < 0,001) e 0,0945 ± 0,0066 e 0,075 ± 0,018 para o estroma (p < 0,001), respectivamente. Além disso, a área de lesão média do Grupo I foi de 69,2 mm2, valor significativamente maior em relação ao Grupo II (49,4 mm2, p = 0,023) e Grupo III (48,6 mm2, p = 0,006). Não houve diferença estatisticamente significante entre os Grupos II e III. CONCLUSãO: A talidomida mostrou-se eficaz na redução da área da lesão e CPI dos implantes de endometriose experimental tanto na dose de 1 mg/kg/dia quanto na dose de 10 mg/kg/dia.

Asunto(s)

Inhibidores de la Angiogénesis/farmacología , Proliferación Celular/efectos de los fármacos , Endometriosis/patología , Endometrio/patología , Talidomida/farmacología , Animales , Biomarcadores/análisis , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Antígeno Nuclear de Célula en Proliferación/análisis , Ratas Wistar

RESUMEN

Abstract Objective To analyze the effect of thalidomide on the progression of endometriotic lesions experimentally induced in rats and to characterize the pattern of cell proliferation by immunohistochemical Proliferating Cell Nuclear Antigen (PCNA) labeling of eutopic and ectopic endometrium. Methods Fifteen female Wistar rats underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium and fixation of a tissue segment to the pelvic peritoneum. Four weeks after, the animals were divided into 3 groups: control (I), 10mg/kg/day (II) and 1mg/kg/day (III) intraperitoneal thalidomide for 10 days. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma analysis. Eutopic and ectopic endometrial tissue was submitted to immunohistochemistry for analysis of cell proliferation by PCNA labeling and the cell proliferation index (CPI) was calculated as the number of labeled cells per 1,000 cells. Results Group I showed a mean CPI of 0.248 ± 0.0513 in the gland and of 0.178 ± 0.046 in the stroma. In contrast, Groups II and III showed a significantly lower CPI, that is, 0.088 ± 0.009 and 0.080 ± 0.021 for the gland (p < 0.001) and 0.0945 ± 0.0066 and 0.075 ± 0.018 for the stroma (p < 0.001), respectively. Also, the mean lesion area of Group I was 69.2mm2, a significantly higher value compared with Group II (49.4mm2, p = 0.023) and Group III (48.6mm2, p = 0.006). No significant difference was observed between Groups II and III. Conclusion Thalidomide proved to be effective in reducing the lesion area and CPI of the experimental endometriosis implants both at the dose of 1mg/kg/day and at the dose of 10 mg/kg/day.

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Resumo Objetivo Analisar o efeito da talidomida na progressão de lesões endometrióticas induzidas experimentalmente em ratas e caracterizar o padrão de proliferação celular pela marcação imunohistoquímica de Antígeno Nuclear de Célula Proliferativa (PCNA) no endométrio eutópico e ectópico. Métodos Quinze ratas Wistar foram submetidas a laparotomia para indução de endometriose por ressecção de um corno uterino, isolamento do endométrio e fixação de um segmento do tecido ao peritônio pélvico. Após quatro semanas, os animais foram divididos em 3 grupos: controle (I), 10 mg/kg/dia (II) e 1 mg/kg/dia (III) de talidomida intraperitoneal por um período de 10 dias. As lesões foram resseccionadas juntamente com o corno uterino oposto para análise da glândula endometrial e do estroma. O tecido endometrial eutópico e ectópico foi submetido à imunohistoquímica para análise da proliferação celular por marcação com PCNA e o índice de proliferação celular (CPI) foi calculado como o número de células marcadas por 1.000 células. Resultados O grupo I apresentou média de CPI de 0,248 ± 0,0513 na glândula e de 0,178 ± 0,046 no estroma. Em contraste, os grupos II e III apresentaram CPI significativamentemenor, isto é, 0,088 ± 0,009 e 0,080 ± 0,021 para a glândula (p < 0,001) e 0,0945 ± 0,0066 e 0,075 ± 0,018 para o estroma (p < 0,001), respectivamente. Além disso, a área de lesãomédia do Grupo I foi de 69,2mm2, valor significativamentemaior em relação ao Grupo II (49,4mm2, p = 0,023) e Grupo III (48,6mm2, p = 0,006). Não houve diferença estatisticamente significante entre os Grupos II e III. Conclusão A talidomida mostrou-se eficaz na redução da área da lesão e CPI dos implantes de endometriose experimental tanto na dose de 1mg/kg/dia quanto na dose de 10 mg/kg/dia.

Asunto(s)

Humanos , Animales , Femenino , Talidomida/farmacología , Inhibidores de la Angiogénesis/farmacología , Modelos Animales de Enfermedad , Endometriosis/patología , Endometrio/patología , Biomarcadores/análisis , Ratas Wistar , Antígeno Nuclear de Célula en Proliferación/análisis , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga

RESUMEN

Thalidomide was originally developed to treat primary neurological and psychiatric diseases. There are reports of anticonvulsant effects of thalidomide in rats and antiepileptic effects in patients. Hence, thalidomide (100, 200 and 400 mg/kg) was herein administered to mice to evaluate possible protection against seizures induced by the systemic administration of neurotoxins: 10 mg/kg of 4-aminopyridine (4-AP), 90 mg/kg of pentylenetetrazol (PTZ), or 380 mg/kg of pilocarpine. The effect of an NO and COX inhibitor (7-NI and ibuprofen, respectively) was also examined. The results show that thalidomide (1) induces the typical sedative effects, (2) has no anticonvulsant effect in mice treated with 4-AP, and (3) has anticonvulsant effect (400 mg/kg) in mice treated with PTZ and pilocarpine. It was found that 7-NI has an anticonvulsant effect in the pilocarpine model and that thalidomide's effect is not enhanced by its presence. However, thalidomide (200 mg/kg) plus 7-NI or ibuprofen tend to have a toxic effect in PTZ model. On the other hand, the combination of thalidomide and 7-NI or ibuprofen protects against pilocarpine-induced seizures. In conclusion, thalidomide did not exert an anticonvulsant effect for clonic-tonic type convulsions (4-AP), but it did so for seizures induced by PTZ and pilocarpine (representing absence seizures and status epilepticus, respectively). NO and prostaglandins were involved in the convulsive process elicited by pilocarpine.

Asunto(s)

Anticonvulsivantes , Neurotoxinas/efectos adversos , Pentilenotetrazol/efectos adversos , Pilocarpina/efectos adversos , Convulsiones/prevención & control , Talidomida/administración & dosificación , Talidomida/farmacología , 4-Aminopiridina/efectos adversos , Enfermedad Aguda , Animales , Modelos Animales de Enfermedad , Quimioterapia Combinada , Ibuprofeno/administración & dosificación , Indazoles/administración & dosificación , Masculino , Ratones Endogámicos , Óxido Nítrico , Convulsiones/inducido químicamente

RESUMEN

BACKGROUND: ß-Blockers reduce temporomandibular joint (TMJ) pain. We asked whether they also reduce TMJ inflammation and, if so, whether this anti-inflammatory effect contributes to its analgesic action. METHODS: We measured many parameters of the inflammatory response after co-administration of the ß-blocker propranolol with the inflammatory agent carrageenan in the TMJ of female rats. We also hypothesized that the activation of ß-adrenoceptors in the TMJ induces nociception mediated, at least in part, by the inflammatory response. To test this hypothesis, we examined the nociceptive response induced by the activation of the ß-adrenoceptors in the TMJ in female rats pretreated with thalidomide and fucoidan. RESULTS: We found that the co-administration of propranolol with carrageenan in the TMJ of female rats significantly reduced several parameters of the inflammatory response induced by carrageenan such as plasma extravasation, neutrophil migration and the release of the pro-inflammatory cytokines TNF-α, IL-1ß and CINC-1. Furthermore, the injection of the ß-adrenergic receptor agonist isoproterenol in the TMJ induced nociception that was significantly reduced by thalidomide, fucoidan and by the co-administration of propranolol but not of the α-adrenergic receptor antagonist phentolamine. CONCLUSIONS: Propranolol has anti-inflammatory effects that contribute to its antinociceptive action in the TMJ of females. SIGNIFICANCE: ß-Blockers have an anti-inflammatory effect on temporomandibular joint (TMJ) that contributes to its analgesic effect. The results of this work suggest that ß-blockers can be used to treat the painful conditions of TMJ, especially when they are associated with an inflammatory process.

Asunto(s)

Antagonistas Adrenérgicos beta/farmacología , Nocicepción/efectos de los fármacos , Propranolol/farmacología , Articulación Temporomandibular/efectos de los fármacos , Antagonistas Adrenérgicos alfa/farmacología , Analgésicos/uso terapéutico , Animales , Antiinflamatorios/uso terapéutico , Anticoagulantes/farmacología , Carragenina/farmacología , Quimiocina CXCL1/efectos de los fármacos , Quimiocina CXCL1/inmunología , Femenino , Inmunosupresores/farmacología , Interleucina-1beta/efectos de los fármacos , Interleucina-1beta/inmunología , Dolor/tratamiento farmacológico , Dimensión del Dolor/efectos de los fármacos , Fentolamina/farmacología , Polisacáridos/farmacología , Ratas , Ratas Wistar , Articulación Temporomandibular/inmunología , Trastornos de la Articulación Temporomandibular/tratamiento farmacológico , Talidomida/farmacología

RESUMEN

Fetal hemoglobin (HbF) induction by hydroxyurea (HU) therapy is associated with decreased morbidity and mortality in sickle cell anemia (SCA) patients, but not all patients respond to or tolerate HU. This provides a rationale for developing novel HbF inducers to treat SCA. Thalidomide analogs have the ability to induce HbF production while inhibiting the release of tumor necrosis factor-alpha. Molecular hybridization of HU and thalidomide was used to synthesize 3- (1,3-dioxoisoindolin-2-yl) benzyl nitrate (compound 4C). In this study, we show that compound 4C increases HbF production in a transgenic SCA mouse model and reduces the production of pro-inflammatory cytokines by SCA mouse monocytes cultured ex vivo. Therefore, compound 4C is a novel drug designed to treat SCA with a unique combination of HbF-inducing and anti-inflammatory properties.

Asunto(s)

Anemia de Células Falciformes/tratamiento farmacológico , Citocinas/metabolismo , Hemoglobina Fetal/biosíntesis , Hidroxiurea , Talidomida , Anemia de Células Falciformes/genética , Anemia de Células Falciformes/metabolismo , Anemia de Células Falciformes/patología , Animales , Citocinas/genética , Modelos Animales de Enfermedad , Hemoglobina Fetal/genética , Hidroxiurea/análogos & derivados , Hidroxiurea/síntesis química , Hidroxiurea/química , Hidroxiurea/farmacología , Inflamación/tratamiento farmacológico , Inflamación/genética , Inflamación/metabolismo , Inflamación/patología , Ratones , Ratones Noqueados , Talidomida/análogos & derivados , Talidomida/síntesis química , Talidomida/química , Talidomida/farmacología

RESUMEN

TRIAL DESIGN: Open-label, randomised, controlled, pilot proof-of-concept clinical trial. METHODS: Participants: Antiretroviral naïve adult males with CD4 count ≥350cells/mm3. INTERVENTIONS: Patients were randomised to receive thalidomide 200mg QD for 3weeks (Thalidomide group) or not (Control group) and followed for 48weeks. OBJECTIVE: We hypothesized that short-term Thalidomide use would reduce HIV related inflammation and HIV replication among antiretroviral naïve HIV infected individuals. OUTCOME: Viral loads, CD4/CD8 counts, ultra-sensitive C-reactive protein (US-CRP), cell activation markers, and plasma lipopolysaccharide (LPS) were analyzed. Randomisation: Unrestricted randomisation. Blinding: No blinding was used. RESULTS: Numbers randomised: Thirty recruited individuals were randomised to Thalidomide (16 patients) or Control (14 patients) groups. Recruitment: Patients were recruited from April 2011 to January 2013. OUTCOME: Viral loads remained stable in both groups. During thalidomide treatment, a decrease in CD4/CD8 ratio (p=0.04), a decrease in CD4 count (p=0.04), an increase in cell activation calculated by the percentage of CD38 +/HLA-DR+ CD8 cells (p<0.05) and an increase in US-CRP (p<0.01) were observed in the Thalidomide group, with all parameters returning to baseline levels after thalidomide interruption. We confirmed that thalidomide increased HIV replication in vitro of 6 of 7 samples from long-term antiretroviral suppressed individuals. HARMS: No class 3/4 adverse events occurred. CONCLUSIONS: Short-term use of thalidomide led to an intense transient increase in T cell activation and inflammation, with a decrease in the CD4+ cell count without changes to the CD8+ cell count. We confirmed that thalidomide acts in vitro as a latency reversal agent and speculate that the in vivo results obtained were due to an increase in HIV replication.

Asunto(s)

Infecciones por VIH/inmunología , Inmunosupresores/farmacología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Talidomida/farmacología , Terapia Antirretroviral Altamente Activa , Recuento de Linfocito CD4 , Estudios de Casos y Controles , Citocinas/sangre , Citocinas/metabolismo , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Humanos , Masculino , Ensayos Clínicos Controlados Aleatorios como Asunto , Receptores CCR5/metabolismo , Receptores CXCR4/metabolismo , Linfocitos T/metabolismo , Resultado del Tratamiento , Carga Viral , Tropismo Viral

RESUMEN

JAK proteins have been linked with survival and proliferation of multiple myeloma (MM) cells; therefore, JAK inhibition could be a therapeutic strategy for MM. We evaluated JAK1 and JAK2 expression in MM patients and the effects of JAK/STAT pathway inhibition on apoptosis, cell cycle, gene and protein expression in RPMI-8226 and U266 MM cell lines. 57% of patients presented overexpression of JAK2 and 27%, of JAK1. After treatment with ruxolitinib and bortezomib, RPMI-8226 and U266 presented 50% of cells in late apoptosis, reduction of anti-apoptotic genes expression and higher number of cells in SubG0 phase. Co-culture with stromal cells protected RPMI-8226 cells from apoptosis, which was reversed by lenalidomide addition. Combination of ruxolitinib, bortezomib and lenalidomide induced 72% of cell death, equivalent to bortezomib, lenalidomide and dexamethasone, combination used in clinical practice. Many JAK/STAT pathway genes, after treatment, had their expression reduced, mainly in RPMI-8226, with insignificant changes in U266. In this scenario, JAK/STAT pathway could pose as a new therapeutic target to be exploited, since it is constitutively active and contributes to survival of MM tumor cells.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Bortezomib/farmacología , Janus Quinasa 1/antagonistas & inhibidores , Janus Quinasa 2/antagonistas & inhibidores , Mieloma Múltiple/tratamiento farmacológico , Pirazoles/farmacología , Factores de Transcripción STAT/metabolismo , Transducción de Señal/efectos de los fármacos , Talidomida/análogos & derivados , Adulto , Anciano , Anciano de 80 o más Años , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Ciclo Celular/efectos de los fármacos , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Técnicas de Cocultivo , Relación Dosis-Respuesta a Droga , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Janus Quinasa 1/genética , Janus Quinasa 1/metabolismo , Janus Quinasa 2/genética , Janus Quinasa 2/metabolismo , Lenalidomida , Masculino , Persona de Mediana Edad , Mieloma Múltiple/enzimología , Mieloma Múltiple/genética , Mieloma Múltiple/patología , Nitrilos , Pirimidinas , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismo , Células del Estroma/patología , Talidomida/farmacología , Factores de Tiempo

RESUMEN

A series of related thalidomide derivatives (2-9) were synthesized by microwave irradiation and evaluated for anti-inflammatory activity. Such activity was assessed in vivo and ex vivo. Compounds 2, 8 and 9 showed the highest levels of inhibition of TNF-α production. On rat paw edema and hyperalgesia assays, compound 9, (1,4-phthalazinedione) demonstrated the highest in vivo anti-inflammatory activity. Thus, compound 9 can be considered as a promising compound to be subjected to further modification to obtain new agents for the treatment of inflammatory diseases.

Asunto(s)

Antiinflamatorios , Talidomida , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Animales , Antiinflamatorios/síntesis química , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéutico , Líquido del Lavado Bronquioalveolar/inmunología , Carragenina , Edema/inducido químicamente , Edema/tratamiento farmacológico , Edema/metabolismo , Pie/patología , Hiperalgesia/inducido químicamente , Hiperalgesia/tratamiento farmacológico , Lipopolisacáridos , Masculino , Ratones Endogámicos BALB C , Peroxidasa/metabolismo , Ratas Sprague-Dawley , Talidomida/análogos & derivados , Talidomida/farmacología , Talidomida/uso terapéutico , Factor de Necrosis Tumoral alfa/inmunología

Asunto(s)

HDL-Colesterol/sangre , Talidomida/análogos & derivados , Talidomida/farmacología , Triglicéridos/sangre , Factor de Necrosis Tumoral alfa/sangre , Animales , HDL-Colesterol/antagonistas & inhibidores , Relación Dosis-Respuesta a Droga , Hipercolesterolemia/sangre , Hipercolesterolemia/tratamiento farmacológico , Masculino , Ratas , Ratas Wistar , Talidomida/uso terapéutico , Triglicéridos/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores

RESUMEN

We investigated the mechanisms of action of immuno-modulatory drug (lenalidomide) on the protein expression of cereblon (CRBN) and their therapeutic targets in the multiple myeloma cell line RPMI8226. The multiple myeloma cell line RPMI8226 was cultured and treated with different concentrations of lenalidomide and bortezomib to determine the proliferation inhibition rate, apoptosis rate, and protein expression of CRBN. The results revealed that both lenalidomide and bortezomib inhibited the proliferation of RPMI8226 and promoted cell apoptosis. However, the protein expression of CRBN decreased signifi-cantly after treatment with lenalidomide, while bortezomib had no effect on the expression of CRBN. We confirmed that CRBN may be a target of lenalidomide.

Asunto(s)

Mieloma Múltiple/metabolismo , Péptido Hidrolasas/metabolismo , Talidomida/análogos & derivados , Proteínas Adaptadoras Transductoras de Señales , Apoptosis/efectos de los fármacos , Western Blotting , Bortezomib/farmacología , Línea Celular Tumoral , Humanos , Lenalidomida , Talidomida/farmacología , Ubiquitina-Proteína Ligasas

RESUMEN

PURPOSE: To develop thalidomide-loaded poly-lactide-co-glycolide implants and evaluate its in vivo release and biological activity against inflammation and angiogenesis after subcutaneous administration. METHODS: Implants were prepared by the hot molding technique and characterized using stereomicroscopy, thermal analysis and X-ray diffraction. Swiss mice, divided in groups 1-3, received a subcutaneous implant containing 25% (w/w), 50% (w/w) or 75% (w/w) of thalidomide, respectively (n=6). The drug levels were determined during a 28-day study period. The toxicity associated with the implants was evaluated by light microscopy. The potential of the developed implant in the inhibition of inflammation and angiogenesis was evaluated in vivo using the sponge model. RESULTS: Thalidomide implant was developed and its characterization proved the stability of the drug and the polymer during preparation. Release profiles in vivo demonstrated an extended release of thalidomide from the implants during the 28 days. Histological evaluation did not show any sign of intense local inflammatory response to the presence of the implants in the subcutaneous pouch. The thalidomide implant reduced the number of vessels and N-acetyl-b-glucosaminidase (NAG) in vivo. CONCLUSION: The biodegradable implants delivered safe doses of thalidomide that were also effective to induce angiogenesis and inflammation regression.

Asunto(s)

Materiales Biocompatibles/química , Inflamación/tratamiento farmacológico , Neovascularización Patológica/tratamiento farmacológico , Talidomida/administración & dosificación , Talidomida/uso terapéutico , Acetilglucosaminidasa/metabolismo , Animales , Rastreo Diferencial de Calorimetría , Modelos Animales de Enfermedad , Femenino , Hemoglobinas/metabolismo , Inflamación/patología , Inyecciones Subcutáneas , Ácido Láctico/química , Ratones , Neovascularización Patológica/patología , Peroxidasa/metabolismo , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Piel/efectos de los fármacos , Piel/patología , Talidomida/farmacología , Difracción de Rayos X

RESUMEN

Thalidomide has shown protective effects in different models of ischemia/reperfusion damage. To elucidate the mechanisms of such protection, this study assessed the effects of thalidomide on the oxidative stress and inflammatory response induced by ischemia/reperfusion episodes in rats. Rats underwent middle cerebral artery occlusion (MCAO) for 2hours. All animals were sacrificed after different reperfusion times. Rats were administered either DMSO or thalidomide (20mg/kg (i.p.)) at different times before or during reperfusion: 1) 1h before reperfusion; the infarct area was measured 2h after reperfusion. 2) 10min before reperfusion and 80min after reperfusion; the infarct area was measured 24h after reperfusion; and 3) 10min before reperfusion and 1h, 24h, 48h, and 68h after reperfusion; the infarct area was measured 72h after reperfusion. Thalidomide reduced the infarct area 24h and 72h after MCAO, and decreased the neurological deficit in all groups with respect to controls. Thalidomide also lowered significantly the number of TUNEL-positive cells, levels of Bax, caspase-3, lipoperoxidation, and pro-inflammatory cytokines, and increased the levels of SOD1, Bcl-2 and pAkt. These results show that thalidomide has neuroprotective effects, apparently due to its anti-apoptotic, anti-oxidant, and anti-inflammatory effects.

Asunto(s)

Inhibidores de la Angiogénesis/farmacología , Isquemia Encefálica/tratamiento farmacológico , Fármacos Neuroprotectores/farmacología , Daño por Reperfusión/tratamiento farmacológico , Talidomida/farmacología , Animales , Apoptosis/efectos de los fármacos , Modelos Animales de Enfermedad , Inflamación/tratamiento farmacológico , Masculino , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar

RESUMEN

A encefalomielite autoimune experimental (EAE) é uma doença inflamatória e desmielinizante do sistema nervoso central (SNC) caracterizada por incapacidades temporárias ou permanentes. A patogênese envolve a reação auto-imune associada com a produção de citocinas pró inflamatórias, tais como o fator de necrose tumoral alfa (TNF-α). Esta citocina está associada com o aumento de radicais livres de oxigênio, como o óxido nítrico, liberados pelas células imunes ativadas. Além de aumentar a inflamação, tanto o fator de necrose tumoral, como o óxido nítrico causam lesão tecidual direta. Este estudo avaliou o efeito da talidomida na progressão clínica da doença, desenvolvimento da reação inflamatória e desmielinização. A expressão tecidual "in situ" do TNF-α e iNOS, uma enzima associada com a produção de óxido nítrico, foi investigada em amostras do SNC obtidos durante o desenvolvimento do modelo de EAE em ratos Lewis. Métodos: Ratos Lewis(n = 30) foram divididos em grupo de controle saudável (I), grupo experimental de encefalomielite autoimune (II) e o grupo tratado com talidomida (III). Os ratos foram monitorizados durante 15 dias para determinação da condição clínica, após este período, os animais foram eutanasiados e as amostras do sistema nervoso central foram obtidas para a realização de estudo histopatológico e imuno-histoquímico Resultados: Todos os animais do grupo II tiveram sintomas relacionados a EAE, enquanto apenas um do grupo tratado talidomida apresentaram alterações clínicas. O estudo histopatológico revelou que as amostras de SNC do grupo II apresentaram áreas de intenso infiltrado inflamatório mononuclear difuso e presença de áreas de desmielinização. No entanto, os animais tratados com talidomida apresentaram ocasionalmente um leve infiltrado inflamatório e bainhas de mielina bem organizadas. Além disso, a expressão de TNF-α e iNOS foram significativamente maiores no grupo II, quando comparado com o grupo tratado com a talidomida. Conclusões: Os resultados considerados em conjunto sustentam a hipótese de que a talidomida inibe a intensidade do processo inflamatório e desmielinização, assim como reduz a produção de mediadores inflamatórios modulando o desenvolvimento da encefalomielite auto-imune experimental em ratos Lewis.

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Experimental autoimmune encephalomyelitis is a inflammatory and demyelinating disease of central nervous system (CNS) characterized by permanents or temporary disabilities. Its pathogenesis involves autoimmune reaction associated with the production of pro inflammatory cytokines such as tumor necrosis factor alpha (TNF-α). This cytokine is associated with increase of reactive oxygen free radicals, such as nitric oxide, released by activated immune cells. Besides enhancing inflammation, both tumor necrosis factor as nitric oxide cause pathologically direct destruction of proteins and enzyme oxidation. This study focuses on clinical disease progression, development of the inflammatory reaction and evaluation axonal myelination . The " in situ" tissue expression of the TNF-α and inducible nitric oxide synthase iNOS ,an enzyme associated with the production of nitric oxide , were also investigated in CNS samples obtained during the development of experimental autoimmune encephalomyelitis model in Lewis rats. Methods: Lewis rats were used to perform the classical model of EAE. The rats ( n=30) were divided into the healthy control group (I), experimental autoimmune encephalomyelitis group (II) and thalidomide treated group (III). The rats were monitored for 15 days for determination of clinical score , after this period , the animals were euthanized and samples were obtained from the central nervous system in which histopatological study and immunohistochemistry for SNC in situ detection of TNF-α and inducible nitric oxide synthase (iNOS) were performed. Results: All animals of group II had symptoms related to experimental encephalomyelitis , while only one of the thalidomide treated group showed clinical changes. The histopatological study revealead that SNC samples of group II presented areas of intense focal and diffuse mononuclear inflammation and the myelin sheaths were scarce and poorly stained. However, thalidomide treated rats presented occasionally a mild perivascular inflammatory infiltrate and myelin sheaths were organized and well evidenced. In addition, the expression of TNF-α and iNOS were significantly higher in the group II when compared with thalidomide treated group. Conclusions: The results taken together support the hypothesis that thalidomide inhibits the intensity of the inflammation and demyelination process and as well as reduces the production of inflammatory mediators influencing the development of experimental autoimmune encephalomyelitis in Lewis rats

Asunto(s)

Animales , Ratas , Talidomida/farmacología , Factor de Necrosis Tumoral alfa/farmacología , Enfermedades Desmielinizantes , Óxido Nítrico Sintasa/farmacología , Encefalomielitis Autoinmune Experimental/patología , Ratas Endogámicas Lew