The antidepressant effect of albiflorin based on TSPO and mechanism / 中国药理学通报

Aim To study the antidepressant effects of albiflorin and its relationship with TSPO(translocator protein 18 ku). Methods Mice were divided into eight groups(control group, chronic unpredictable stress group, fluoxetine group, albiflorin low, medium, high dose groups, PK11195 group, PK11195+ albiflorin high dose group)based on the data of the behavioral tests conducted to assess the antidepressant-like effects of albiflorin. After the behavioral tests Western blot and ELISA were conducted to evaluate the TSPO expression, progesterone and allopregnanolone level in hippocampus of mice. Results In the behavioral tests, there were significant differences between the model group and the control group, which indicated that the model was successfully established. The positive drug and albiflorin in different dose groups could reverse the effects of the model group, and PK11195 could reverse the effects of paeoniflorin in high dose group. The results of Western blot and ELISA showed that the TSPO expression, progesterone and allopregnanolone level in the model group significantly decreased. The positive drug and albiflorin groups with different doses could reverse the effects of the model group, and PK11195 could reverse the effects of the high dose group. Conclusions Albiflorin has significant antidepressant and antianxiety effects on CUS mice by TSPO, which provides experimental basis for the further study on the antidepressant effects and mechanism of albiflorin in the future.

El cáncer bucal y su relación con la proteína translocadora mitocondrial de 18kda / Oral cancer and its relation with the 18kda mitochondrial translocator protein

Resumen El cáncer bucal posee una alta incidencia y mortalidad a nivel global. A pesar de los avances en el diagnóstico y el pronóstico de esta enfermedad, aún se mantiene una baja tasa de supervivencia de 5 años, lo cual hace necesario el estudio de métodos diagnósticos que sean capaces de detectar la enfermedad en estadios tempranos. Es por esto que avances en proteómica e inmunohistoquímica, han permitido identificar diversos biomarcadores, entre ellos la proteína translocadora (TSPO) mitocondrial de 18kDa, la cual está involucrada en diversos procesos celulares, como el transporte de colesterol, la proliferación celular y la apoptosis. Se ha reportado la presencia de valores alterados de la TSPO en diversos tipos de cáncer, así como la presencia de la TSPO en saliva y tejido de sujetos con cáncer bucal, lo cual representa una oportunidad para entender el proceso de la carcinogénesis bucal e identificar nuevas alternativas para el diagnóstico de esta enfermedad. La presente revisión de tema tiene como objetivo presentar aspectos teóricos en relación con la TSPO como un biomarcador a estudiar en sujetos con cáncer bucal, considerando su implicación en los procesos de apoptosis celular y participación en el estrés oxidativo.

Abstract Oral cancer has a high incidence and mortality rate globally. Despite the advances in the diagnosis and prognosis of this disease, a 5 years survival rate still remains, which makes it necessary to study diagnostic methods capable to detect the disease in early stages. That is why advances in proteomics and immunohistochemistry had allowed the identification of various biomarkers, including the 18 kDa mitochondrial translocator protein (TSPO), which is involved in some cellular processes, such as cholesterol transport, cellular proliferation and apoptosis. It has been reported the altered TSPO values in various types of cancer, as well as the presence of TSPO in saliva of subjects with oral cancer, which represents an opportunity to understand the oral carcinogenic process and identify new alternatives for the diagnosis of this disease. The objective of this review is to present theoretical aspects related to TSPO as a biomarker to study in subjects with oral cancer, considering its implication in the apoptosis mechanism and participation in oxidative stress.

Resumo O câncer bucal apresenta alta incidência e mortalidade ao nível mundial. Apesar dos avanços no diagnóstico e prognóstico da doença, uma baixa taxa de sobrevivência de 5 anos ainda é mantida, o que requer o estudo de métodos de diagnóstico capazes de detectar a doença nos estágios iniciais. é por isso que os avanços na proteômica e imuno-histoquímica identificaram diversos biomarcadores, incluindo a proteína translocadora (TSPO) 18 kDa mitocondriais, que está envolvida em diversos processos celulares, tais como o transporte de colesterol, a proliferação celular e a apoptose. Tem sido relatada a presença de valores alterados da TSPO em diferentes tipos de câncer, assim como a presença da TSPO em saliva e tecidos de pacientes com câncer bucal, o que representa uma oportunidade para entender o processo da carcinogênese bucal e identificar novas alternativas para o diagnóstico desta doença. A presente revisão de literatura tem como objetivo apresentar aspectos teóricos em relação ao uso da TSPO como um biomarcador a ser estudado em pacientes com câncer bucal, considerando seu envolvimento nos processos de apoptose celular e participação no estresse oxidativo.

Mecanismos de modulação da ANXA1 sobre a função da proteína translocadora (TSPO) em células da glia (OU) Mecanismos de modulação da anexina A1 sobre a função da proteína translocadora em células da glia / Annexin A1 modulation mechanism on the expression of the Translocator Protein (TSPO) in BV2 cells

A inflamação no sistema nervoso central (SNC) está envolvida na gênese de uma série de doenças neurodegenerativas, sendo assim, compreender o processo inflamatório nessas circunstâncias se torna essencial para propor novas abordagens terapêuticas. Sabemos que a Anexina A1 (ANXA1) e o receptor TSPO são dois moduladores importantes da neuroinflamação. Enquanto se sabe que a ANXA1 possui propriedades antiinflamatórias, o papel do TSPO ainda não está esclarecido. Desta forma, este projeto avaliou a atuação da ANXA1 sobre a expressão do TSPO em linhagem de células da microglia (BV2), e sua conexão com o receptor Toll-like receptor-4 (TLR4) em BV2 ativada pelo lipopolisacarídeo de E.coli (LPS). Os resultados obtidos mostram que o tratamento de BV2 com LPS induz a expressão de TSPO, dependente de ativação de TLR4, através das vias da molécula adaptadora do fator de diferenciação mielóide 88 (MyD88) e do fator nuclear κB (NFκB). O tratamento com ANXA1 recombinante induz um perfil antiinflamatório em células BV2 estimuladas com LPS, por reduzir a secreção de citocinas proinflamatórias e, ao mesmo tempo, aumentar secreção de citocinas antiinflamatórias. A exposição com ANXA1 ainda impede o aumento da expressão de TSPO induzida pelo LPS. Mostramos também que esta ação da ANXA1 é dependente da interação com o receptor de peptídeo formilado (FPR2). Adicionalmente, o silenciamento de TSPO em células BV2 predispõe essas células a um perfil ativado exacerbando a secreção do fator de necrose tumoral (TNFα) em resposta ao LPS, o que não pode ser revertido pelo tratamento com ANXA1 recombinante. Em conjunto, os resultados expõe a relação existente entre ANXA1 e TSPO em micróglia ativada pelo LPS, mostrando que a ANXA1 9 modula negativamente a expressão do TSPO. Ademais, o silenciamento de TSPO inibiu a fagocitose de neurônios apoptóticos, o que ainda sugere a participação do TSPO na eferocitose

Inflammation in the Central Nervous System (CNS) is involved in the genesis of a number of neurodegenerative diseases, so understanding the inflammatory process in these circumstances is essential to proposal new therapeutic approaches. We know that Annexin A1 (ANXA1) and the TSPO receptor are two important modulators of neuroinflammation. While it is known that ANXA1 has anti-inflammatory properties, the role of TSPO has not yet been clarified. Thus, this project evaluated the interference of ANXA1 on the expression of TSPO in microglia cell line (BV2), and its connection with the Toll-like receptor-4 receptor (TLR4) in BV2 activated by E. coli lipopolysaccharide LPS). The results show that the treatment of BV2 with LPS induces the expression of TSPO, dependent on activation of TLR4, through the pathways of the adapter molecule of myeloid differentiation factor 88 (MyD88) and nuclear factor κB (NFκB). Treatment with recombinant ANXA1 induces an anti-inflammatory profile in LPS-stimulated BV2 cells, by reducing the secretion of proinflammatory cytokines and, at the same time, increasing secretion of anti-inflammatory cytokines. Exposure with ANXA1 still prevents the increase of LPS-induced TSPO expression. We also show that this action of ANXA1 is dependent on the interaction with the formylated peptide receptor (FPR2). In addition, TSPO silencing in BV2 cells predisposes these cells to an activated profile exacerbating secretion of tumor necrosis factor (TNFα) in response to LPS, which can not be reversed by treatment with recombinant ANXA1. Together, the results show the relationship between ANXA1 and TSPO in LPS activated microglia, showing that ANXA1 negatively modulates TSPO 11 expression. In addition, TSPO silencing inhibited the phagocytosis of apoptotic neurons, which still suggests the participation of TSPO in eferocytosis

Translocator protein 18ku: A potential new therapeutic target for anxiety in the future / 国际药学研究杂志

Anxiety is one of the psychiatric disorders. However, the pathogenesis of anxiety is still uncertain. Translocator protein (18×103, 18ku) (TSPO) ligands have been proved useful in the treatment of neurological and psychiatric disorders, especially anxiety. TSPO Ligands are promising candidates for fast-acting anxiolytic drugs with less severe side effects than benzodiazepines. This review summaries TSPO molecular structure, distribution, and its function of treating anxiety and illustrates the advantage of TSPO ligands in treating anxiety and its application prospect.

Systematic Analysis of Translocator Protein 18 kDa (TSPO) Ligands on Toll-like Receptors-mediated Pro-inflammatory Responses in Microglia and Astrocytes

Translocator protein 18 kDa (TSPO) is a mitochondrial protein highly expressed on reactive microglia and astrocytes, and is considered as a biomarker for neurodegeneration and brain damage, especially neuroinflammation. Toll-like receptors (TLRs) are closely related with inflammatory responses of microglia and astrocytes and these signaling pathways regulate neuroinflammation. Previous reports have identified the anti-inflammatory effects of TSPO ligands, however study of their effects in relation to the TLR signaling was limited. Here, we investigated the effects of five representative TSPO ligands on microglia and astrocytes following activation by various TLR ligands. Our results show that TSPO ligands reduce the pro-inflammatory response elicited by the TLR ligands with more profound effects on microglia than astrocytes.

Efeitos dos ligantes de TSPO sobre a migração leucocitária: participação de receptores de glicorticóides / Effects of TSPO ligands on leukocyte migration: role of glucocorticoids receptors

Além dos receptores no sistema nervoso central (SNC), acoplados a receptores GABAA (CBR - central benzodiazepine receptor), outros sítios periféricos de ligação para os benzodiazepínicos (BDZ) foram descritos e denominados de "periféricos" (PBR peripheral benzodiazepine receptor) ou TSPO - Translocator protein. Sua função como modulador do sistema imune tem sido proposta, com a participação, pelo menos em parte, de glicocorticóides endógenos (GE). No entanto, os mecanismos da modulação dos TSPO sobre o processo inflamatório e a relação destes receptores com os GE ainda não estão totalmente estabelecidos. Desta forma, este trabalho visou investigar os efeitos de dois ligantes sintéticos de TSPO, um benzodiazepínico, Ro5-4864, e uma isoquinolona carboxamida, PK 11195, e suas relações com os GE, sobre os eventos da migração leucocitária. Para tanto, ratos Wistar machos foram tratados in vivo com RU 38486 (antagonista de receptor de GE) ou com veículo e, adicionalmente, os ligantes de TSPO, Ro5-4864 e/ou PK 11195, foram aplicados topicamente sobre a microcirculação do mesentério dos animais para avaliação do comportamento rolling e aderência de leucócitos, por microscopia intravital, em condições basais ou após estimulação pelo peptídeo formil-metionil-leucilfenilalanina (fMLP). Leucócitos periféricos foram coletados e tratados in vitro com os ligantes de TSPO para avaliação da expressão de moléculas de adesão, quimiotaxia e influxo de cálcio em condições basais ou após estimulação pelo fMLP. Ainda, culturas primárias de células endoteliais da microcirculação de ratos foram tratadas in vitro com os ligantes de TSPO para avaliação da expressão de moléculas de adesão em condições basais ou após estimulação pelo lipopolisacarídeo de E.coli (LPS). Os resultados obtidos mostraram que: 1) o ligante de TSPO, Ro5-4864, alterou a ação do fMLP, uma vez que inibiu a diminuição do rolling, o aumento de células aderentes, a clivagem de L-selectina, o aumento de expressão de PECAM-1 (platelet endothelial cell adhesion molecule), a quimiotaxia e o influxo de cálcio em neutrófilos; não afetou a expressão de PECAM-1 e ICAM-1 (intracellular cell adhesion molecule) induzida pelo LPS em células endoteliais; 2) o PK 11195 aumentou o influxo de cálcio e a quimiotaxia de neutrófilos induzido pelo fMLP; 3) a co-administração dos ligantes de TSPO, Ro5-4864 e PK 11195, reverteu os efeitos do Ro5-4864 sobre a expressão de L-selectina e PECAM-1, o influxo de cálcio e a quimiotaxia de neutrófilos. A possível inter-relação dos TSPO e GE foi verificada uma vez que o Ro5-4864 inibiu o aumento do número de leucócitos rolling e aderidos à parede vascular in vivo causado pelo tratamento com RU 38486; inibiu o aumento da expressão de L-selectina em neutrófilos e de ß2-integrina presente em linfócitos e de PECAM-1 em neutrófilos. Ainda, foi verificado que neutrófilos possuem expressões de TSPO maiores que linfócitos e células endoteliais e que esta expressão é aumentada em neutrófilos pela ação do fMLP. Em conjunto, os resultados obtidos neste trabalho sugerem que os ligantes de TSPO modulam diferentemente as funções de adesão e mobilidade de leucócitos e de adesão da célula endotelial e que uma relação alostérica de agonismo/antagonismo entre estes pode ser sugerida. Adicionalmente, os resultados mostram uma possível relação entre os TSPO e os receptores de citoplasmáticos de GE no controle da interação leucócito-endotélio

Besides central benzodiazepine receptor (CBR) found on central nervous system (CNS) coupled to GABAA, another binding site for benzodiazepines was described and named peripheral benzodiazepine receptor (PBR) or Translocator protein (TSPO). The mainly function described to this TSPO is steroidogenesis. Additionally, its function as an immune modulator has been proposed with the involvement, at least in part, of endogenous glucocorticoids (EG). However, mechanisms of TSPO modulation on inflammatory process and the relation between the receptor and EG has not been totally established. This study aimed investigates effects of two synthetic TSPO ligands, a benzodiazepine, Ro5-4864, and an isoquinoline carboxamide, PK 11195, and their relation with EG on leukocyte migration. For this purpose, male Wistar rats were in vivo treated with RU 38486 (EG antagonist receptor) or vehicle and, in addition, TSPO ligands, Ro5-4864 and/or PK 11195, were topically applied on animal mesentery microcirculation to evaluate leukocyte rolling and adhesion, by intravital microscopy, under basal conditions or after stimulation by N-formyl-methionine-leucine-phenylalanine (fMLP). In addition, peripheral leukocytes were collected and in vitro treated with TSPO ligands to evaluate adhesion molecules expression, chemotaxis and calcium influx. Endothelial cells obtained from cremaster muscle were in vitro treated with TSPO ligands to evaluate adhesion molecules expression under basal conditions or after stimulation by lipopolysaccharide from E.coli (LPS). Our results showed that: 1) TSPO ligand, Ro5-4864, altered the fMLP action since TSPO ligand inhibited the decrease of rolling, the increase of adherent cells, L-selectin cleavage, the increase of PECAM-1 expression, neutrophil chemotaxis and calcium influx; did not affect expression of PECAM-1 and ICAM-1 induced by LPS on endothelial cells; 2) PK 11195 increased fMLP-induced neutrophil calcium influx and chemotaxis; 3) co-administration of TSPO ligands, Ro5-4864 and PK 11195, reverted Ro5-4864 effects on L-selectin and PECAM-1 expression, neutrophil and calcium influx, as well as PECAM-1 and ICAM-1 expression on endothelial cells; 3) a possible inter-relation between TSPO and EG was observed since Ro5-4864 inhibited the increase of in vivo leukocyte rolling and adhesion induced by RU 38486 in vivo treatment; it inhibited the increase of neutrophil L-selectin, lymphocyte ß2-integrin and leukocyte PECAM-1 expressions; 4) It was showed that neutrophil have more TSPO than lymphocytes and endothelial cells, and this expression is increased by fMLP stimulation. Taken together, our results suggest that TSPO ligands differently modulate leukocyte adhesion and motility functions and endothelial cells adhesion, and further, they suggest an allosteric agonist/antagonist relation. Additionally, they showed a relation between TSPO and EG cytoplasmic receptor on the control of leukocyte-endothelial interaction

TSPO:new nomenclature for the peripheral benzodiazepine receptor and its role in the neuropsycopharmacology / 中国药理学通报

The early characterization of the diazepam-binding sites outside the brain led to their assignment as peripheralbenzodiazepine receptors, or PBRs, to distinguish them from the central benzodiazepine receptor.Although PBR is a widely used and accepted name in the scientific community,recent data regarding the structure and molecular function of this protein increasingly support renaming it to represent more accurately its subcellular role (or roles) and putative tissue-specific function (or functions). Translocator protein (18 ku,TSPO), is proposed as a new name, regardless of the subcellular localization of the protein. This review deals with the pharmacological, structural and molecular characterization of the PBR and its role in the neuropshcopharmacology.

Synthesis and anxiolytic effects of 2-aryl imidazo1,2-apyridine-3- acetamide derivatives / 国际药学研究杂志

Objective: To search for new compounds with better water-solubility and higher antianxiety activities. Methods: A series of 2-arylimidazo[1,2-a] pyridine-3-acetamide derivitives were designed and synthesized. The anxiolytic activities were evaluated by BZDR competitive binding assay in vitro and the elevated-plus maze test in mice, the structure-activity relationship (SAR) has been studied. Results and Conclusion: Twenty-eight new compounds have been synthesized. Their structures were confirmed by 1H NMR and MS. According to the results of BZDR affinity test, compounds I1, I 8, I10, I13, I19 showed as good affinity as the positive control (Ro5-4864). The corresponding inhibition was 87%, 89%, 85%, 89% and 76% respectively at the concentration of 100 nmol/L, while that of Ro5-4864 was 82%. I8 and I10, which display better water-solubility and better BZDR affinity in vitro, show significant antianxiety effects in vivo.