The effect of 2-amino-3-carboxymuconate-6-semialdehyde decarboxylase gene overexpression in the kynurenine pathway on the expression levels of indoleamine 2,3-dioxygenase 1 and interferon-γ in inflammatory conditions: an in vitro study

Background The kynurenine pathway (KP) can be involved in the pathogenesis of neurodegenerative diseases and excessive neurotoxic metabolite production. This study aimed to evaluate the effects of overexpression of murine 2-amino-3-carboxymuconate-6-semialdehyde decarboxylase ( Acmsd ) gene in infla...

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Published in:Molecular biology reports 2022-02, Vol.49 (2), p.1103-1111
Main Authors: Rostaminejad, Marzieh, Rostami, Abdolmohamad, Behzad-Behbahani, Abbas, Dehbidi, Gholam Reza Rafiei, Kalantari, Tahereh
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animal Anatomy
Animal Biochemistry
Animals
Biomedical and Life Sciences
Carboxy-Lyases - genetics
Carboxy-Lyases - metabolism
Gene expression
Green fluorescent protein
Histology
Indoleamine-Pyrrole 2,3,-Dioxygenase - genetics
Indoleamine-Pyrrole 2,3,-Dioxygenase - metabolism
Inflammation
Interferon-gamma - genetics
Interferon-gamma - metabolism
Kidney - metabolism
Kynurenine - metabolism
Life Sciences
Lipopolysaccharides
Liver - metabolism
Macrophages
Mice
Morphology
Neural coding
Neurodegenerative diseases
Neurotoxicity
Original Article
Plasmids
Polyethyleneimine
RAW 264.7 Cells
Transfection
Tryptophan 2,3-dioxygenase
γ-Interferon
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Summary:Background The kynurenine pathway (KP) can be involved in the pathogenesis of neurodegenerative diseases and excessive neurotoxic metabolite production. This study aimed to evaluate the effects of overexpression of murine 2-amino-3-carboxymuconate-6-semialdehyde decarboxylase ( Acmsd ) gene in inflammatory conditions in RAW 264.7 cell line to present more information about the effect of this gene on inflammatory conditions and the KP cycle. Methods and results The coding sequence of the Acmsd gene was cloned into pCMV6-AC-IRES-GFP expression vector with a green fluorescent protein (GFP) marker. To simulate inflammatory conditions, RAW 264.7 macrophage cells were stimulated by Lipopolysaccharide (LPS) 24 h before transfection, and transfected by Polyethyleneimine (PEI) with constructed plasmids expressing the Acmsd gene. The effect of Acmsd gene expression level on murine Interferon-gamma ( Ifn-γ ) and murine Indoleamine 2,3-dioxygenase 1 ( Ido1 ) gene expression level was investigated by Real-Time PCR. According to the results of this study, good transfection efficiency was observed 72 h after transfection, and Acmsd expression level increased 29-fold (P 
ISSN:0301-4851
1573-4978
DOI:10.1007/s11033-021-06935-4