Autofagia mediada pelo canabidiol em modelo de Doença de Parkinson em Caenorhabditis elegans
Data
2024-06-28
Autores
Kano, Rodrigo Issamu 
Orientadores
Pereira, Gustavo José da Silva
Tipo
Trabalho de conclusão de curso
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O envelhecimento é o fator de risco primário para doenças neurodegenerativas como a Doença de Parkinson (DP). Muitos indivíduos acometidos pela DP recebem tratamento farmacológico paliativo que ajudam a controlar as manifestações clínicas da doença, porém, sem inibir sua progressão. Neste contexto, embora o canabidiol (CBD) tenha sido cada vez mais estudado na DP, devido a seus efeitos no sistema nervoso central e por apresentar efeitos neuroprotetores, o seu mecanismo de ação ainda não está totalmente elucidado. Muitas evidências apontaram para um papel do CDB na indução da autofagia em vários modelos celulares e animais. Sabe-se que a deficiência da autofagia leva ao acúmulo citosólico de agregados proteicos ubiquitinados, que são características importantes de processos neurodegenerativos, como os corpos de Lewis na DP, que são formados principalmente pela proteína α-sinucleína. Desta forma, o objetivo deste projeto foi estudar a ação do CBD em modelos in vivo da Doença de Parkinson, avaliando a modulação da termotolerância, o papel da autofagia e uma possível via de ação HLH-30/TFEB. Foram utilizados modelos de C. elegans transgênicos, para expressão de GFP::LGG-1 e GFP::HLH-30, para a avaliação das proteínas LC3 e TFEB, respectivamente. A microscopia de fluorescência foi utilizada para avaliar os efeitos do CBD na formação de pontuações e a translocação de HLH-30/TFEB para o núcleo para avaliar uma possível indução da autofagia através da ativação do HLH-30/TFEB após o tratamento de CBD. Verificou-se também a possível modulação do CBD na longevidade pelo ensaio de termotolerância em diferentes concentrações, CBD 0,1 µM, CBD 0,5 µM, CBD 1 µM, CBD 5 µM e CBD 100 µM. Os dados obtidos por microscopia de fluorescência sugeriram a ativação do fluxo autofágico pela via TFEB após o tratamento com CBD a 100 μM por 2 horas. Dessa forma, esse estudo contribui para uma melhor compreensão sobre os potenciais alvos terapêuticos canabinoides e como perspectiva futura um possível papel neuroprotetor.
Aging is the primary risk factor for most neurodegenerative diseases as Parkinson disease (PD). Several individuals affected by this disease receive palliative treatment that helps to control clinical manifestation, however, without inhibiting disease progression. Cannabidiol (CBD) has been studied in PD research, due to its effect on the central nervous system and for showing neuroprotective effect, however the underline mechanism is still unknown. Several evidences have demonstrated a potential role for CBDinduced autophagy via the master regulator of autophagy Transcription Factor EB (TFEB). Autophagy has been shown to be an important process in the context of neurodegenerative diseases, by either reducing or excessively increasing their autophagic activity. It is already known that the autophagy deficiency leads to aberrant aggregation of ubiquitinated proteins, an important characteristic of the neurodegenerative processes, as Lewis body on the Parkinson’s diseases, formed mostly of αsynuclein protein. For this purpose, the aim of this project is evaluate the effect of CBD in a in vivo model of Caenorhabditis elegans for PD, assessing the modulation of termoresistence at heatshock assays and autophagy activation though HLH30/TFEB translocation to the nucleus, in this context. Transgenic C. elegans models were used for the expression of GFP::LGG1 and GFP::HLH30 to the, respectively, autophagy and HLH30/TFEB evaluation. Fluorescence microscopy techniques were used to evaluate the effects of CBD on the punctae formation and HLH30 translocation and so to evaluate a possible autophagy induction by HLH30/TFEB activation after CBD treatment. Our results showed that termoresistence at the heatshock assay by different concentration comparing to the CTR group, CBD 0,1 μM, CBD 0,5 μM, CBD 1 μM, CBD 5 μM and CBD 100 μM. Furthermore, an autophagy flux activation and HLH30/TFEB translocation by 100 uM CBD treatment for 2 hours. Therefore, this study may contribute to a better understanding of the potential cannabinoid therapeutic targets and as a future perspective their possible neuroprotective effects.
Aging is the primary risk factor for most neurodegenerative diseases as Parkinson disease (PD). Several individuals affected by this disease receive palliative treatment that helps to control clinical manifestation, however, without inhibiting disease progression. Cannabidiol (CBD) has been studied in PD research, due to its effect on the central nervous system and for showing neuroprotective effect, however the underline mechanism is still unknown. Several evidences have demonstrated a potential role for CBDinduced autophagy via the master regulator of autophagy Transcription Factor EB (TFEB). Autophagy has been shown to be an important process in the context of neurodegenerative diseases, by either reducing or excessively increasing their autophagic activity. It is already known that the autophagy deficiency leads to aberrant aggregation of ubiquitinated proteins, an important characteristic of the neurodegenerative processes, as Lewis body on the Parkinson’s diseases, formed mostly of αsynuclein protein. For this purpose, the aim of this project is evaluate the effect of CBD in a in vivo model of Caenorhabditis elegans for PD, assessing the modulation of termoresistence at heatshock assays and autophagy activation though HLH30/TFEB translocation to the nucleus, in this context. Transgenic C. elegans models were used for the expression of GFP::LGG1 and GFP::HLH30 to the, respectively, autophagy and HLH30/TFEB evaluation. Fluorescence microscopy techniques were used to evaluate the effects of CBD on the punctae formation and HLH30 translocation and so to evaluate a possible autophagy induction by HLH30/TFEB activation after CBD treatment. Our results showed that termoresistence at the heatshock assay by different concentration comparing to the CTR group, CBD 0,1 μM, CBD 0,5 μM, CBD 1 μM, CBD 5 μM and CBD 100 μM. Furthermore, an autophagy flux activation and HLH30/TFEB translocation by 100 uM CBD treatment for 2 hours. Therefore, this study may contribute to a better understanding of the potential cannabinoid therapeutic targets and as a future perspective their possible neuroprotective effects.
Descrição
Citação
KANO, Rodrigo Issamu. Autofagia mediada pelo canabidiol em modelo de Doença de Parkinson em Caenorhabditis elegans. 2024. 42f. Trabalho de Conclusão de Curso (Graduação em Biomedicina) – Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP). São Paulo, 2024.