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Exploring the role of carbon monoxide as a modulator of microglia neurotrophism
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RESUMO: A comunicação celular é essencial para a manutenção da integridade e
homeostase neuronal no Sistema Nervoso Central (SNC). A microglia é responsável
por várias funções no SNC, tais como o refinamento sináptico, patrulha dos
microambientes e, principalmente, pela resposta imunitária e inflamação, vitais para
a sobrevivência e bom funcionamento neuronal.
A microglia desempenha diversas funções devido à comunicação bidirecional
com outros tipos de celulares, incluindo os neurónios. A regulação da microglia é
crucial, uma vez que ações exacerbadas podem levar a uma inflamação
descontrolada e promover danos irreversíveis. O monóxido de carbono (CO) é uma
molécula endógena com propriedades citoprotectoras e anti-inflamatórias
comprovadas quando presente em níveis baixos no SNC, incluindo a limitação da
produção microglial de agente pró-inflamatórios.
No entanto, a influência do CO na função basal da microglia permanece
desconhecida. O objetivo principal deste estudo foi compreender a ação do CO na
modulação da neuroprotecção microglial, em condições fisiológicas.
Foi estabelecido um protocolo de meio condicionado usando linhas celulares
de BV2-microglia e CAD-neurónios. Os neurónios incubados com meio de microglia
apresentaram alterações morfológicas e aumento de viabilidade sugerindo que o
secretoma da microglia induz neuroprotecção. De forma a compreender a influência
do CO nesta resposta migroglial foi realizado um tratamento com uma nova molécula
libertadora de CO (ALF-826).
Verificámos que o tratamento com CO aumentou a expressão do factor
neurotrófico BDNF e a secreção da Interleucina-10 na microglia. Do mesmo modo, o
CO também aumentou a expressão ectonucleotidase microglial CD73, indicando um
possível envolvimento da sinalização purinérgica, em particular da adenosina.
Concluímos que o CO reforça a neuroprotecção através da modulação do
secretoma microglial. O progresso do conhecimento sobre a função reguladora do
CO na microglia pode ser considerado um avanço de uma nova potencial aplicação
do CO como procedimento preventivo através da melhoria do neurotrofismo da
microglia.
ABSTRACT: Cellular communication is essential for the maintenance of neuronal functional integrity and environmental homeostasis in the Central Nervous System (CNS). Microglia are responsible for various functions in the CNS such as synaptic pruning, patrol of the microenvironments and, most importantly, immune response and inflammation, vital for neuronal survival and tissue homeostasis. Microglia is capable of performing very diverse and distinct roles due to the bi-directional communication with other cell types, including neurons. Microglial activity needs to be under tight regulation since exacerbated actions can lead to uncontrolled inflammation and promote irreversible damage. Carbon monoxide (CO) is an endogenous gasotransmitter with already proven cytoprotective and anti-inflammatory effects when present at low levels in the CNS, including the limitation of pro-inflammatory microglial output. Nevertheless, the influence of CO in basal microglial function remains unexplored. Therefore, the main objective of our study was to understand the effect of CO in the microglial neuroprotection modulation, under physiological conditions. Firstly, in order to achieve this purpose, a conditioned media protocol was established with BV2-microglia and CAD-neuron cell lines. Neurons incubated with microglia medium showed morphological changes and increased viability suggesting that the microglial secretome induces neuroprotection. Secondly, to understand how CO modulates this response, a new CO-releasing molecule (ALF-826) was used to treat microglia and their neuroprotective effect was enhanced. We have found that CO treatment increased the expression of neurotrophic factor BDNF and Interleukin-10 secretion in microglia. Likewise, CO also enhanced microglial ectonucleotidase CD73 expression, indicating that purinergic signaling, in particular adenosine might be involved. In conclusion, CO modulates microglial secretome towards neuroprotection. Furthering the knowledge regarding how CO regulates microglia function could be a step forward towards a new potential CO application as a preventive procedure by modulation and improvement of microglia neurotrophism.
ABSTRACT: Cellular communication is essential for the maintenance of neuronal functional integrity and environmental homeostasis in the Central Nervous System (CNS). Microglia are responsible for various functions in the CNS such as synaptic pruning, patrol of the microenvironments and, most importantly, immune response and inflammation, vital for neuronal survival and tissue homeostasis. Microglia is capable of performing very diverse and distinct roles due to the bi-directional communication with other cell types, including neurons. Microglial activity needs to be under tight regulation since exacerbated actions can lead to uncontrolled inflammation and promote irreversible damage. Carbon monoxide (CO) is an endogenous gasotransmitter with already proven cytoprotective and anti-inflammatory effects when present at low levels in the CNS, including the limitation of pro-inflammatory microglial output. Nevertheless, the influence of CO in basal microglial function remains unexplored. Therefore, the main objective of our study was to understand the effect of CO in the microglial neuroprotection modulation, under physiological conditions. Firstly, in order to achieve this purpose, a conditioned media protocol was established with BV2-microglia and CAD-neuron cell lines. Neurons incubated with microglia medium showed morphological changes and increased viability suggesting that the microglial secretome induces neuroprotection. Secondly, to understand how CO modulates this response, a new CO-releasing molecule (ALF-826) was used to treat microglia and their neuroprotective effect was enhanced. We have found that CO treatment increased the expression of neurotrophic factor BDNF and Interleukin-10 secretion in microglia. Likewise, CO also enhanced microglial ectonucleotidase CD73 expression, indicating that purinergic signaling, in particular adenosine might be involved. In conclusion, CO modulates microglial secretome towards neuroprotection. Furthering the knowledge regarding how CO regulates microglia function could be a step forward towards a new potential CO application as a preventive procedure by modulation and improvement of microglia neurotrophism.
Descrição
Palavras-chave
Microglia Modulação da Neuroprotecção Microglial Monóxido de Carbono Citometria de Fluxo Microglia Modulation of Microglia Neuroprotection Carbon Monoxide Flow Cytometry