RESUMO
Plasma cytokine concentration in patients with Parkinson's disease and mutation in GBA gene, in patients with sporadic Parkinson's disease, and in healthy volunteers were measured by ELISA and multiplex analysis. In patients with Parkinson's disease and mutation in GBA gene, elevated plasma concentrations of IL-1ß and TNFα were revealed by ELISA in comparison with both controls and patients with sporadic form of Parkinson's disease. Multiplex analysis revealed enhanced secretion of IL-1ß, IL-2, IFNγ and reduced plasma levels of monocyte chemoattractant protein-1 (MCP-1) in patients with Parkinson's disease and mutation in GBA gene (in comparison with other groups) and increased plasma levels of IL-13 (only in comparison with the healthy volunteers). Our results support the hypothesis that the concentrations of inflammatory mediators are increased in patients with Parkinson's disease and mutation in GBA gene.
Assuntos
Glucosilceramidase/genética , Mutação , Doença de Parkinson/genética , Idoso , Estudos de Casos e Controles , Quimiocina CCL2/sangue , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Expressão Gênica , Glucosilceramidase/sangue , Glucosilceramidase/imunologia , Humanos , Inflamação , Interferon gama/sangue , Interferon gama/genética , Interferon gama/imunologia , Interleucina-13/sangue , Interleucina-13/genética , Interleucina-13/imunologia , Interleucina-1beta/sangue , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-2/sangue , Interleucina-2/genética , Interleucina-2/imunologia , Masculino , Pessoa de Meia-Idade , Doença de Parkinson/sangue , Doença de Parkinson/imunologia , Doença de Parkinson/patologia , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
The adipose tissue is considered today as an endocrine organ in which tissue-specific regulation of gene expression plays a key role in the processes of development of obesity and comorbidities, such as diabetes and cardiovascular disease. The present study is focused on ITLN1, PPARã and TNFá gene expression in intra-abdominal adipose tissue and its effect on the serum levels of omentin 1 and TNFa in individuals with different body mass. It has been shown that serum TNFa level is significantly higher in the subgroup of patients with overweight and obesity (BMI ≥ 25 kg/m2) as compared to individuals with normal body weight (BMI < 25 kg/m2)( p < 0.03). We have demonstrated that the expression level of the PPARã gene is positively correlated with the ITLN1 gene expression level in the intra-abdominal adipose tissue (r = 0.516, p = 0.020). Serum level of omentin 1 positively correlates with PPARã mRNA and protein levels in intra-abdominal adipose tissue (r = 0.550, p < 0.05 and r = 0.581, p < 0.03, respectively). For the subgroup of patients with overweight and obesity, we have shown negative correlation of the level of TNFá mRNA with PPARã and ITLN1 mRNA levels was shown (r = 0.549, p < 0.05 and r = 0.475, p < 0.05, respectively). This study is the first to show a correlation relationship between PPARã gene expression level in the intra-abdominal adipose tissue and the expression and secretion levels of omentin 1.
Assuntos
Citocinas/biossíntese , Regulação da Expressão Gênica , Gordura Intra-Abdominal/metabolismo , Lectinas/biossíntese , Obesidade/metabolismo , PPAR gama/biossíntese , Fator de Necrose Tumoral alfa/biossíntese , Adulto , Feminino , Proteínas Ligadas por GPI/biossíntese , Humanos , Gordura Intra-Abdominal/patologia , Masculino , Pessoa de Meia-Idade , Obesidade/patologiaRESUMO
Parkinson's disease (PD) is a common neurodegenerative disorder. Disease symptoms correlate with the degeneration of dopaminergic neurons in substantia nigra pars compacta. A number of factors are supposed to take part in PD pathogenesis including alpha-synuclein aggregation, oxidative stress, mitochondrial dysfunction and apoptosis, although the precise molecular mechanism of neudegeneration remains unknown. PD is generally a sporadic neuro- logical disorder, however, rare monogenic forms have been described during previous 15 years. Despite of the fact that mutations in the leucine-rich repeat kinase (LRRK2) gene are the most common cause of inherited forms of PD known today, the mechanisms by which mutations in the LRRK2 gene lead to disease remain unclear. It's difficult to understand the signaling pathways, regulated by LRRK2 in the absence of its clear physiological substrates. The G2019S substitution was shown to be the most common in mutations' spectrum of the LRRK2 gene in different populations which makes it easier to reveal patients with LRRK2-associated PD. In this review LRRK2 influence on protein aggregation, cytoskeletal dynamics, apoptosis rate and inflammatory response is discussed. Groups of patients with inherited forms of PD with known etiology are worth to be included into investigations. It could promote our un- derstanding of the mechanisms of neurodegeneration in more common sporadic cases.
Assuntos
Apoptose/genética , Mutação de Sentido Incorreto , Doença de Parkinson , Agregação Patológica de Proteínas , Proteínas Serina-Treonina Quinases , Transdução de Sinais/genética , Substituição de Aminoácidos , Animais , Citoesqueleto/genética , Citoesqueleto/metabolismo , Citoesqueleto/patologia , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Humanos , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Parte Compacta da Substância Negra/metabolismo , Parte Compacta da Substância Negra/patologia , Agregação Patológica de Proteínas/genética , Agregação Patológica de Proteínas/metabolismo , Agregação Patológica de Proteínas/patologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismoRESUMO
Last data demonstrated that exonic variants of LRRK2 (p.G2019S, p.M1646T) may affect the catalytic activity of lysosomal enzyme glucocerebrosidase (GCase) probably through the phosphorylation of Rab10 protein. We aimed to evaluate an association of LRRK2 exonic variants previously associated with alteration of phosphorylation levels for Rab10Thr73 with PD risk in Russian population and analyze an impact of p.G2019S mutation and selected LRRK2 variants on lysosomal hydrolase activities. LRRK2 variants were determined by full sequencing of LRRK2 in 508 PD patients and 470 controls from Russian population. Activity of lysosomal enzymes (glucocerebrosidase (GCase), alpha-galactosidase A (GLA), acid sphingomyelinase (ASMase) and concentrations of their corresponded substrates (hexosylsphingosine (HexSph), globotriaosylsphingosine (LysoGb3), lysosphingomyelin (LysoSM), respectively) were estimated in 211 PD patients and 179 controls by liquid chromatography with tandem mass spectrometry (LC-MS-MS) in dry blood spots. p.M1646T and p.N2081D were associated with PD (OR = 2.33, CI 95%: 1.1215 to 4.8253, p = 0.023; OR = 1.89, 95%CI: 1.0727 to 3.3313, p = 0.028, respectively) in Russian population. An increased LysoGb3 concentration was found in p.G2019S and p.N2081D LRRK2 carriers among PD patients compared to both PD patients and controls (p.G2019S: p = 0.00086, p = 0.0004, respectively; p.N2081D: p = 0.012, p = 0.0076, respectively). A decreased ASMase activity in p.G2019S LRRK2 carriers among PD patients (p = 0.014) was demonstrated as well. Our study supported possible involvement of LRRK2 dysfunction in an alteration of sphingolipid metabolism in PD.
Assuntos
Doença de Parkinson , Humanos , Doença de Parkinson/genética , Glucosilceramidase/genética , Glucosilceramidase/metabolismo , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/genética , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina/metabolismo , Mutação , Esfingolipídeos , LisossomosRESUMO
Mutations in the Leucine Reach Repeat Kinase 2 (LRRK2) gene are the most frequent cause of familial Parkinson's disease (PD). Although the precise physiological and pathological role of LRRK2 is unclear, a direct link between mutant LRRK2 and apoptosis has been suggested. Using flow cytometric analysis (PI+Annexin V(FITC)) we showed increased spontaneous apoptosis of peripheral blood lymphocytes in patients with LRRK.2-associated PD compared to controls after 24 (P < 0.016) and 48 (P < 0.031 ) h of incubation (5 % CO2, 37 degrees C). We found the increased FAS mRNA level in peripheral blood lymphocytes of patients with LRRK2-associated PD compared to controls (P < 0.05) and to sporadic PD (sPD) (P < 0.002). Significant difference in FAS expression between patients with LRRK2-associated PD and controls remained after three years and was detected after 1 and 24 h during lymphocyte incubation (P < 0.03 and 0.05, respectively). Increased spontaneous lymphocytes apoptosis along to increased FAS expression in patients with LRRK2-associated PD suggest that LRRK2 mutations may lead to the activation of extrinsic apoptotic way.
Assuntos
Apoptose/genética , Linfócitos/patologia , Doença de Parkinson , Proteínas Serina-Treonina Quinases/genética , Idoso , Estudos de Casos e Controles , Análise Mutacional de DNA , Feminino , Citometria de Fluxo , Humanos , Serina-Treonina Proteína Quinase-2 com Repetições Ricas em Leucina , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Doença de Parkinson/genética , Doença de Parkinson/patologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Reação em Cadeia da Polimerase em Tempo Real , Federação Russa , Receptor fas/genéticaRESUMO
The synucleinopathies are a group of neurodegenerative diseases characterized by the oligomerization of alpha-synuclein protein in neurons or glial cells. Recent studies provide data that ceramide metabolism impairment may play a role in the pathogenesis of synucleinopathies due to its influence on alpha-synuclein accumulation. The aim of the current study was to assess changes in activities of enzymes involved in ceramide metabolism in patients with different synucleinopathies (Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA)). The study enrolled 163 PD, 44 DLB, and 30 MSA patients as well as 159 controls. Glucocerebrosidase, alpha-galactosidase, acid sphingomyelinase enzyme activities, and concentrations of the corresponding substrates (hexosylsphingosine, globotriaosylsphingosine, lysosphingomyelin) were measured by liquid chromatography tandem-mass spectrometry in blood. Expression levels of GBA, GLA, and SMPD1 genes encoding glucoceresobridase, alpha-galactosidase, and acid sphingomyelinase enzymes, correspondently, were analyzed by real-time PCR with TaqMan assay in CD45 + blood cells. Increased hexosylsphingosine concentration was observed in DLB and MSA patients in comparison to PD and controls (p < 0.001) and it was associated with earlier age at onset (AAO) of DLB (p = 0.0008). SMPD1 expression was decreased in MSA compared to controls (p = 0.015). Acid sphingomyelinase activity was decreased in DLB, MSA patients compared to PD patients (p < 0.0001, p < 0.0001, respectively), and in MSA compared to controls (p < 0.0001). Lower acid sphingomyelinase activity was associated with earlier AAO of PD (p = 0.012). Our data support the role of lysosomal dysfunction in the pathogenesis of synucleinopathies, namely, the pronounced alterations of lysosomal activities involved in ceramide metabolism in patients with MSA and DLB.
Assuntos
Doença por Corpos de Lewy , Atrofia de Múltiplos Sistemas , Doença de Parkinson , Sinucleinopatias , Ceramidas , Humanos , Doença por Corpos de Lewy/metabolismo , Atrofia de Múltiplos Sistemas/patologia , Doença de Parkinson/patologia , Esfingolipídeos , Esfingomielina Fosfodiesterase , alfa-Galactosidase , alfa-Sinucleína/metabolismoRESUMO
Parkinson's disease (PD) is a multifactorial neurodegenerative disease. To date, genome-wide association studies have identified more than 70 loci associated with the risk of PD. Variants in the GBA gene encoding glucocerebrosidase are quite often found in PD patients in all populations across the world, which justifies intensive investigation of this gene. A number of biochemical features have been identified in patients with GBA-associated Parkinson's disease (GBA-PD). In particular, these include decreased activity of glucocerebrosidase and accumulation of the glucosylceramide substrate. These features were the basis for putting forward a hypothesis about treatment of GBA-PD using new strategies aimed at restoring glucocerebrosidase activity and reducing the substrate concentration. This paper discusses the molecular and genetic mechanisms of GBA-PD pathogenesis and potential approaches to the treatment of this form of the disease.
RESUMO
Lysosomal integral membrane protein-2 (LIMP-2), encoded by the SCARB2 gene, is the specific lysosomal receptor for glucocerebrosidase enzyme. Association between rs6812193 and rs68250047 of SCARB2 with PD has been shown in genetic studies, including large genome-wide association studies. The aim of the current study was to determine whether rs6812193 and rs8475 are associated with PD in Russia. rs6812193 and rs8475 were genotyped in a total of 604 PD patients (65 PD patients with positive (fPD) and 539 PD patients with negative family history (sPD)) and 413 controls and also in 17 patients with PD associated with GBA mutations (PD-GBA) and 18 asymptomatic GBA mutation carriers (GBA-Carriers). SCARB2 expression was measured by real-time PCR in CD45+ blood cells in part of individuals in the studied groups. No linkage disequilibrium was shown between rs6812193 and rs8475 in Russian population. Increased PD risk for TT variant of rs8475 (OR = 2.02; p < 0.001) was found in sPD patients but not in fPD. rs6812193 and rs8475 were not associated with age at onset (AAO) of PD. SCARB2 expression level was decreased in GBA-PD patients and GBA-Carriers compared to PD patients (padjusted = 0.02, padjusted = 0.003, respectively) and GBA-Carriers compared to controls (padjusted = 0.013) with no significant difference in PD patients and controls. SCARB2 expression was not modified with rs6812193 and rs8475. In conclusion, rs8475 was associated with PD status. rs6812193 and rs8475 are not genetic modifier of AAO of PD and do not influence on SCARB2 mRNA level in CD45+ blood cells in studied groups. SCARB2 expression could be modified with GBA mutations and is independent of PD status.
Assuntos
Proteínas de Membrana Lisossomal/genética , Doença de Parkinson/genética , Doença de Parkinson/patologia , Receptores Depuradores/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Predisposição Genética para Doença , Variação Genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Doença de Parkinson/sangue , Polimorfismo de Nucleotídeo Único , Federação RussaRESUMO
Immune response may play a pivotal role in the pathogenesis of the common synucleinopathy as Parkinson's disease (PD) and could be mediated with the accumulation of neurotoxic alpha-synuclein. There is limited evidence for immune response in another synucleinopathy as dementia with Lewy bodies (DLB). Recent data suggest that immune response may contribute to cognitive impairment. We aimed to estimate plasma cytokine profile in patients with synucleinopathies with dementia (PD dementia (PDD), DLB). Plasma cytokine levels (interferon-gamma (IFN-gamma), interleukin (IL)-4 (IL-4), IL-6, IL-10, tumor necrosis factor alpha (TNF-alpha), monocyte chemoattractant protein-1 (MCP-1)). were estimated in 16 patients with DLB, 19 patients with PDD, 28 patients with PD without dementia (PD) and 19 individuals without neurological disorders (controls) using Luminex array system. Cognitive status was assessed with the Mini-Mental State Examination (MMSE). TNF-alpha and IL-6 plasma levels were elevated in patients with synucleinopathies with dementia (DLB, PDD) compared to controls and IL-10 plasma level was increased in PDD compared to controls (p < 0.05). IFN-gamma levels were decreased in PD and PDD patients compared to controls (p < 0.001, p = 0.026, respectively) and in PD patients than in DLB patients (p = 0.032). Patients with PD, PDD, and DLB were characterized by increased plasma levels of MCP-1 compared to controls (p < 0.001). At the same time, no differences in TNF-alpha, IL-10, IL-6 plasma levels in PD patients compared to controls were found. Our study demonstrated more pronounced immune response in synucleinopathies associated with dementia compared to PD without demetia.
Assuntos
Citocinas/sangue , Demência/etiologia , Sinucleinopatias/imunologia , Idoso , Idoso de 80 Anos ou mais , Quimiocina CCL2/sangue , Demência/sangue , Demência/imunologia , Feminino , Humanos , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-6/sangue , Doença por Corpos de Lewy/sangue , Doença por Corpos de Lewy/imunologia , Testes de Estado Mental e Demência , Pessoa de Meia-Idade , Doença de Parkinson/sangue , Doença de Parkinson/complicações , Doença de Parkinson/imunologia , Sinucleinopatias/sangue , Sinucleinopatias/complicações , Fator de Necrose Tumoral alfa/sangueRESUMO
Mutations in the GBA and SMPD1 genes, which lead to the development of lysosomal storage diseases, are high risk factors for Parkinson's disease and dementia with Lewy bodies. We screened the mutations in the GALC and CLN3 genes in patients with Parkinson's disease and control subjects. A heterozygous CLN3 mutation (del 1.02 kb) carrier with clinical features of the unusual extrapyramidal syndrome was identified. A role of CLN3 mutations in the development of neurodegenerative disorders is discussed.
Assuntos
Doenças dos Gânglios da Base/genética , Glicoproteínas de Membrana/genética , Chaperonas Moleculares/genética , Deleção de Sequência , Idoso , Feminino , Testes Genéticos , Heterozigoto , Humanos , Leucodistrofia de Células Globoides/genética , Masculino , Doença de Parkinson/genética , Linhagem , SíndromeRESUMO
Tissue specific expression of genes encoding cholesterol transporters ABCA1 and ABCG1 as well as genes encoding the most important transcriptional regulators of adipogenesis - LXRa, LXRb, PPARg and RORa has been investigated in intraabdominal adipose tissue (IAT) samples.A direct correlation between the content of ABCA1 and ABCG1 proteins with RORa protein level (r=0.480, p<0.05; r=0.435, p<0.05, respectively) suggests the role of the transcription factor RORa in the regulation of IAT ABCA1 and ABCG1 protein levels. ABCA1 and ABCG1 gene expression positively correlated with obesity indicators such as body mass index (BMI) (r=0.522, p=0.004; r=0.594, p=0.001, respectively) and waist circumference (r=0.403, p=0.033; r=0.474, p=0.013, respectively). The development of obesity is associated with decreased IAT levels of RORa and LXRb mRNA (p=0.016 and p=0.002, respectively). These data suggest that the nuclear factor RORa can play a significant role in the regulation of cholesterol metabolism and control IAT expression of ABCA1 and ABCG1, while the level of IAT LXRb gene expression may be an important factor associated with the development of obesity.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/metabolismo , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Gordura Abdominal/metabolismo , Obesidade/metabolismo , Transportador 1 de Cassete de Ligação de ATP/genética , Membro 1 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Adulto , Composição Corporal , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Obesidade/genéticaRESUMO
A link between lysosomal storage diseases (LSDs) and neurodegenerative disorders associated with accumulation of presynaptic protein alpha-synuclein has been shown. Particularly, Gaucher disease (GD) patients with a deficiency of the lysosomal enzyme glucocerebrosidase (GBA) and carriers of GBA mutations are at increased risk of Parkinson's disease (PD). It remains unclear whether this link is due to increased alpha-synuclein oligomerization. Here we show that level of oligomeric alpha-synuclein form, associated with PD development, is increased in plasma of GD patients (n=41, median=22.9pg/mL, range1.57-444.58pg/mL; controls (n=40, median=6.02pg/mL, range 1.05-103.14pg/mL, p<0.0001). This difference is absent in GD patients receiving enzyme replacement therapy (ERT) for more than 5 years. Moreover, the levels of alpha-synuclein oligomers in plasma are also higher in patients with other LSDs (Niemann-Pick type C, Krabbe disease, Wolman disease) compared to the median value in controls. Therefore, we suggest that mutations in the GBA gene and at least in several other LSDs genes may be associated with an increase in oligomeric alpha-synuclein in plasma. ERT applied for recovering of GBA functions in GD treatment might decrease formation of plasma oligomeric alpha-synuclein.