Evaluation of ApaI and FokI polymorphism of VDR gene and functional characterization in patients with fibromyalgia / Avaliação do polimorfismo ApaI e FokI do gene VDR e caracterização funcional em pacientes com fibromialgia

Abstract Introduction: Fibromyalgia (FM) is a syndrome of unknown origin characterized by several symptoms, and although its pathogenesis has not been completely elucidated, it seems to be related to inflammatory path-ways and neurochemical changes in the brain. Objective: To evaluate the association between BsmI, ApaI and FokI polymorphisms of the vitamin D receptor (VDR) gene, their polymorphisms, and clinical variables in women with and without FM. Methods: This is a case-control study composed of a group of 53 women with FM and another with 40 women without the disease. The McGill Pain Questionnaire, Fibromyalgia Impact Questionnaire, Pain Visual Analogue Scale and the sit-up test were applied. Real-time PCR was performed to analyze the ApaI and FokI polymorphism. Results: There was a statistical association between race, comorbidity and FM, where 78.4% of the individuals were white and had FM (p < 0.002) and 96.1% had some comorbidity (p < 0.001). Seventy-six point five percent (76.5%) of patients with FM underperformed in the sit-up test (p < 0.001). There was also an association between the genotypic and allele frequencies of the VDR and FM gene Apal and FokI polymorphisms (p < 0.001). In the VDR gene ApaI polymorphism, the CC genotype exhibited a higher frequency in women with FM, the C allele for the Apal polymorphism was 3.33 times more likely, and the FokI polymorphism was 10.9 times more likely to develop FM (p < 0,0001). Conclusion: Women with C allele for ApaI polymorphism are 3.33 times more likely to have FM (95%CI = 1.58-7.02; p = 0.0024), and in FokI polymorphism, the prevalence of T allele is 10.9 times greater (95% CI = 4.76-25.38; p < 0.0001). No significant associations were found in relation to BsmI polymorphism and frequency alleles (p = 0.062 and p = 0.078, respectively).

Resumo Introdução: A fibromialgia (FM) é uma síndrome de origem desconhecida caracterizada por diversos sintomas, e embora sua patogênese não tenha sido completamente elucidada, parece estar relacionada às vias inflamatórias e alterações neuroquímicas no cérebro. Objetivo: Avaliar a associação entre os polimorfismos BsmI, ApaI e FokI do gene do receptor da vitamina D (VDR), seus polimorfismos e variáveis clínicas em mulheres com e sem FM. Métodos: Trata-se de um estudo caso-controle composto por um grupo de 53 mulheres com FM e outro com 40 mulheres sem a doença. Foram aplicados o Questionário de Dor de McGill, Questionário de Impacto da Fibromialgia, Escala Visual Analógica da Dor e o teste de sentar. A PCR em tempo real foi realizada para analisar o polimorfismo ApaI e FokI. Resultados: Houve associação estatística entre raça, comorbidade e FM, onde 78,4% dos indivíduos eram brancos e apresentavam FM (p < 0,002) e 96,1% tinham alguma comorbidade (p < 0,001). Setenta e seis vírgula cinco por cento (76,5%) dos pacientes com FM tiveram desempenho inferior no teste de abdominais (p < 0,001). Também houve associação entre as frequências genotípicas e alélicas dos polimorfismos Apal e FokI do gene VDR e FM (p < 0,001). No polimorfismo ApaI do gene VDR, o genótipo CC apresentou maior frequência em mulheres com FM, o alelo C para o polimorfismo Apal foi 3,33 vezes mais provável, e o polimorfismo FokI teve 10,9 vezes mais chance de desenvolver FM (p < 0,0001). Conclusão: Mulheres com alelo C para polimorfismo ApaI têm 3,33 vezes mais chance de ter FM (IC 95% = 1,58-7,02; p = 0,0024), e no polimorfismo FokI, a prevalência do alelo T é 10,9 vezes maior (IC 95% = 4,76-25,38; p < 0,0001). Não foram encontradas associações significativas em relação ao polimorfismo BsmI e alelos de frequência (p = 0,062 e p = 0,078, respectivamente).

MicroRNA-146a polymorphism is not associated with cardiovascular disease in the elderly / Polimorfismo MicroRNA 146a não está associado a doenças cardiovasculares em idosos

Abstract Introduction: Cardiovascular diseases (CVD) figure among the most significant causes of morbidity and mortality in the world and, among genetic factors, the literature has demonstrated the crucial role of miRNAs and the relationship of physical activity with this pathology. Objective: To investigate the relationship between the functional capacity of exercise, the level of physical activity, and the polymorphism in the miRNA-146a gene in elderly individuals with and without CVD. Methods: This study, developed in a city in the southern region of Brazil, is characterized as cross-sectional. The sample for this study comprised 342 participants, aged 60 or over. The following aspects were analyzed: anthropometric characteristics, genetic profiles, diagnosis of CVD, functional capacity, and the level of physical activity. Results: A statistically significant association was observed between CVD and body mass index (BMI) (א² = 14.278; p = 0.0003), and 40.6% of elderly individuals with CVD were obese, while 31.5% of the normally developed elderly participants presented normal BMI. However, the genotype frequencies (p = 0.546; א² = 1.211) and 6MWT (p = 0.311; א² = 1.025) did not show a statistically signifi-cant association with CVD. Conclusion: Our results suggest that the polymorphism in the miRNA-146A (rs2910164) and functional capacity are not associated with CVD in the elderly. However, the BMI did demonstrate an association with this disease.

Resumo Introdução: As doenças cardiovasculares (DCV) figuram entre as causas mais significativas de morbimortalidade no mundo e, dentre os fatores genéticos, a literatura tem demonstrado o papel crucial dos miRNAs e a relação da atividade física com essa patologia. Objetivo: Investigar a relação entre a capacidade funcional do exercício, o nível de atividade física e o polimorfismo no gene miRNA-146a em idosos com e sem DCV. Métodos: Este estudo, desenvolvido em um município da região sul do Brasil, caracteriza-se como transversal. A amostra deste estudo foi composta por 342 participantes, com idade igual ou superior a 60 anos. Foram analisados ​​os seguintes aspectos: características antropométricas, perfis genéticos, diagnóstico de DCV, capacidade funcional e nível de atividade física. Resultados: Observou-se associação estatisticamente significativa entre DCV e índice de massa corporal (IMC) (א² = 14,278; p = 0,0003), sendo que 40,6% dos idosos com DCV eram obesos, enquanto 31,5% dos idosos normalmente desenvolvidos apresentaram IMC normal. No entanto, as frequências genotípicas (p = 0,546; א² = 1,211) e TC6 (p = 0,311; א² = 1,025) não mostraram associação estatisticamente significante com DCV. Conclusão: Os resultados do presente estudo sugerem que o polimorfismo no miRNA-146A (rs2910164) e a capacidade funcional não estão associados à DCV em idosos; no entanto, o IMC demonstrou associação com essa doença.

Ultrasonic therapy modulates the expression of genes related to neovascularization and inflammation in fibroblasts / Terapia ultrassônica modula a expressão de genes relacionados à neovascularização e inflamação em fibroblastos

Abstract Introduction: In the rehabilitation of musculoskeletal injuries, ultrasound is widely used in clinical practice. Objective: To evaluate the effects of pulsed ultrasonic therapy on the viability and modulation of genes involved in inflammation (IL-6) and neovascularization (VEGF) processes of L929 fibroblast cells. Methods: For irradiation with ultrasound the cells were subdivided into groups: G1 (without irradiation), G2 (0.3 W/cm2-20%) and G3 (0.6 W/cm2-20%), with periods of treatment at 24, 48 and 72 hours. The cell viability assay was analyzed by the MTT method and gene modulation was analyzed by RT-qPCR method. Results: After the comparative analysis between groups, only G2 and G3 (48-hour) presented statistically significant differences in relation to the control. In relation to the gene expression, the selection of the groups analyzed was delimited according to the comparative analysis of the values obtained by the MTT test. After the achievement of RT-qPCR, it could be observed that in G2 the amount of VEGF gene transcripts increased by 1.125-fold compared to endogenous controls, and increased 1.388-fold in G3. The IL-6 gene, on the other hand, had its transcripts reduced in both G2 (5.64x10-9) and G3 (1.91x10-6). Conclusion: Pulsed ultrasound in L929 fibroblasts showed a significant biostimulatory effect in the 48-hour period, with increased cell viability, and the same effect in the modulation of gene expression related the neovascularization and inflammation, mediating the acceleration of the tissue repair cascade.

Resumo Introdução: Na reabilitação de lesões musculoesqueléticas, o ultrassom é amplamente utilizado na prática clínica. Objetivo: Avaliar os efeitos da terapia ultrassônica pulsada sobre a viabilidade e modulação de genes envolvidos nos processos de inflamação (IL-6) e neovascularização (VEGF) de fibroblastos L929. Métodos: Para irradiação com ultrassom, as células foram subdivididas em grupos: G1 (sem irradiação), G2 (0,3 W/cm2-20%) e G3 (0,6 W/cm2-20%), com períodos de tratamento de 24, 48 e 72 horas. O ensaio de viabilidade celular foi analisado pelo método MTT e a modulação gênica pelo método RT-qPCR. Resultados: Após a análise comparativa entre os grupos, apenas G2 e G3 (48 horas) apresentaram diferenças estatisticamente significantes em relação ao controle. Em relação à expressão gênica, a seleção dos grupos analisados ​​foi delimitada de acordo com a análise comparativa dos valores obtidos pelo teste MTT. Após a obtenção do RT-qPCR, pôde-se observar que no G2 a quantidade de transcritos do gene VEGF aumentou 1,125 vezes em relação aos controles endógenos e 1,388 vezes no G3. O gene IL-6, por outro lado, teve seus transcritos reduzidos tanto no G2 (5,64x10-9) quanto no G3 (1,91x10-6). Conclusão: O ultrassom pulsado em fibroblastos L929 apresentou efeito bioestimulador significativo no período de 48 horas, com aumento da viabilidade celular, e o mesmo efeito na modulação da expressão gênica relacionou neovascularização e inflamação, mediando a aceleração da cascata de reparação de tecidos.

Functional status and severity of osteoarthritis in elderly is associated to the polymorphism of TNFA gene

Abstract Background: Osteoarthritis (OA) is a major musculoskeletal disease with high prevalence in the elderly. The study of genetic polymorphisms of inflammatory mediators involved in OA may contribute to the elucidation of the complex pathophysiology of this disease and identification of susceptibility individuals. Aim: This study aimed to evaluate the association between polymorphism at tumor necrosis factor alpha gene (SNP - 308 G/A TNFA) with presence, severity and functional status of osteoarthritis in elderly. Methods: This study was characterized as case-control and encompassed 257 physically independent elderly (Mean Age: 68.55 ± 5.2; Minimum age: 60 and Maximum age: 82) were recruited. After this selection, the groups were divided in: 92 elderly individuals with osteoarthritis (case group) and 165 without the disease (control group). Methods: The individuals were genotyped by the TaqMan real-time PCR system. The subjects were classified based on the degree of radiological impairment according to the criteria of Kellgren-Laurence and regarding functional impairment using the WOMAC and LEQUESNE questionnaires. Results: TNFA gene polymorphic individuals (subjects harboring allele A) are more affected by OA (χ2 = 8.7, p = 0.003), once they have major radiological lesion both in hip (Fisher-Freeman-Halton Test = 3.9, p = 0.04) and knee (Fisher- Freeman-Halton Test = 4.0, p = 0.04) as well as worse functional status assessed by the Lequesne questionnaire (Mann- Whitney, p = 0.04). At the multivariate analysis, after adjustment for age, gender, body mass index, the presence of rare allele for TNFA (allele A) increases the susceptibility to OA development [OR: 1.87 (95% CI: 1.1 —3.2)]. Conclusion: We conclude that the SNP - 308 G/A of TNFA gene may affect osteoarthritis susceptibility, severity and functional status of individuals with osteoarthritis.

Lower limb muscle strength and serotonin receptor gene polymorphism as factors associated in women with fibromyalgia

Abstract Background: Fibromyalgia (FM) is a chronic pain syndrome characterized by generalized skeletal muscle chronic pain. Its etiology is not well defined, because there are several factors that may trigger it such as physical and/or emotional stresses, or a genetic susceptibility, involving serotonergic, dopaminergic and catecholaminergic paths. The objective of this study was to investigate the association between the strength of the lower limb, genetic polymorphism of the serotonin receptor gene HTR2a in women with fibromyalgia. Methods: In this observational study of case-control type 48 women were evaluated who belonged to the group with FM (52 ± 12 years) and 100 women in the control group (58±11 years). Socio demographic and anthropometric data were collected and peripheral blood samples for DNA extraction; genotypic analyzes were performed by means of PCR in real time by TaqMan® system. The lower limb muscle strength was assessed through the test of sitting down and standing up for 30 s. The chi-square test or Fischer Exact was used for possible associations among the variables; the t-test for independent samples was used to compare the averages among the groups; the value of significance adopted was 5%. Results: There was an association between the polymorphism of the HTR2A gene with FM, demonstrating that carriers of the genotype GG have 24.39 times more likely to develop the syndrome (IC95% 5.15-115.47; p = 0.01). It was observed an association between FM and the test to sit and stand up demonstrating that women with fibromyalgia have lower limb muscle strength ( p = 0.01). The study showed that the white race has 3.84 times more likely to develop FM (p = 0.01). Conclusion: The results of this study suggest that women of Caucasian ethnicity with GG genotype or G allele presented greater risk of developing fibromyalgia and that these patients have lower limb muscle strength compared to the control group.