IL-10 and ET-1 as biomarkers of rheumatic valve disease / IL-10 e ET-1 como biomarcadores de doença valvar reumática

Objective: To evaluate the immunological profile and gene expression of endothelin-1 (ET-1) in mitral valves of patients with rheumatic fever originated from a reference service in cardiovascular surgery. Methods: This was a quantitative, observational and cross-sectional study. Thirty-five subjects (divided into four groups) participated in the study, 25 patients with chronic rheumatic heart disease and ten control subjects. The mean age of the sample studied was 34.5 years. Seventeen of them (48.58%) were male and 18 (51.42%) were female. Inflammatory cytokines (TNF-α, IL-4 and IL-10) were measured and ten mitral valves of patients who underwent first valve replacement were collected for determination of gene expression of endothelin-1 by real time PCR. Results: Among the groups studied (patients vs. controls), there was a statistically significant difference in IL-10 levels (P=0.002), and no differences in other cytokines. Expression of endothelin-1 was observed in 70% of samples. Quantitatively, average of ET-1 expression was 62.85±25.63%. Conclusion: Inflammatory cytokine IL-10 participates in the maintenance of chronicity of rheumatic fever in patients who underwent valve replacement and those who are undergoing medical treatment. The expression of endothelin-1 in heart valve lesions in patients undergoing mitral valve replacement confirms its association with inflammatory activity in rheumatic fever. .

Objetivo: Avaliar o perfil imunológico e a expressão gênica de endotelina-1 em valvas mitrais de pacientes com febre reumática, originados de um serviço de referência em cirurgia cardiovascular. Métodos: Este foi um estudo quantitativo, observacional e transversal. Trinta e cinco indivíduos (divididos em quatro grupos) participaram do estudo, 25 deles com doença cardíaca reumática crônica, além de 10 controles. A média de idade da amostra estudada foi de 34,5 anos. Dezessete (48,58%) dos indivíduos eram homens, e 18 (51,42%) eram mulheres. Foram medidas algumas citocinas inflamatórias (TNF-α, IL-4 e IL-10) e coletadas 10 valvas mitrais de pacientes que se submeteram a primeira troca valvar para determinação da expressão gênica de endotelina-1 pelo PCR real-time. Resultados: Entre os grupos estudados (pacientes e controles), observou-se diferença estatisticamente significante em relação aos níveis de IL-10 (P=0,002), sem diferenças nas outras citocinas. Em relação à endotelina-1, foi observada sua expressão em 70% das amostras. Quantitativamente, a expressão média de endotelina-1 foi de 62,85±25,63%. Conclusão: A citocina inflamatória IL-10 participa da manutenção da cronicidade da febre reumática em pacientes que se submeteram a troca valvar e naqueles que estão em tratamento médico. A expressão de endotelina-1 nas lesões em valvas cardíacas de pacientes que foram submetidos à troca valvar mitral confirma sua relação com a atividade inflamatória na febre reumática. .

Gene expression of endothelin receptors in replaced rheumatic mitral stenotic valves / Expressão gênica de receptores de endotelina em valvas mitrais reumáticas estenóticas substituídas

OBJECTIVES: Rheumatic fever is a highly prevalent disease in Brazil, and it poses a major public health problem. It is the leading cause of acquired heart disease in childhood and adolescence. The aim of this study was to evaluate the gene expression of ET-3 and its receptors, in replaced rheumatic mitral valves. METHODS: We studied the gene expression of endothelin-3 (ET-3) and its receptors, endothelin receptor A and endothelin receptor B (ETr-A and ETr-B), in the rheumatic mitral valves of 17 patients who underwent valve replacement surgery. The samples also underwent a histological analysis. RESULTS: Our data showed that almost all patients, regardless of individual characteristics such as gender or age, expressed the endothelin receptor genes, but did not express the genes for ET-3. In quantitative analysis, the ETr-A/GAPDH mean ratio was 33.04 ± 18.09%; while the ETr-B/GAPDH mean ratio was 114.58 ± 42.30%. Regarding histopathological individual features, the frequency of fibrosis is 100%, 88.23% of mononuclear infiltrate, 52.94% of neovascularization, 58.82% of calcification and absence of ossification. CONCLUSION: The presence of receptors ETr-A and ETr-B in rheumatic mitral valves suggests its interaction with the system of circulating endothelins, particularly ETr-B (known for acting in the removal of excess endothelin) detected in a greater proportion, which could explain the lack of expression of endothelin in rheumatic mitral valve, process to be elucidated.

OBJETIVOS: A febre reumática é uma doença altamente prevalente no Brasil, e representa um importante problema de saúde pública. É a principal causa de cardiopatia adquirida na infância e adolescência. O objetivo deste estudo foi avaliar a expressão gênica de ET-3 e seus receptores, em valvas mitrais reumáticas substituídas. Métodos: Estudamos a expressão gênica de endotelina-3 (ET-3) e de seus receptores, receptor da endotelina A e receptor da endotelina B (ETr-A e ETr-B), nas valvas mitrais reumáticas de 17 pacientes que se submeteram à cirurgia de troca valvar. As amostras também foram submetidas à análise histológica. RESULTADOS: Nossos dados mostraram que praticamente todos os pacientes, independentemente de características individuais, como sexo ou idade, expressaram os genes de receptores de endotelina, porém não expressaram os genes para ET-3. Na análise quantitativa, a média da proporção ETr-A/GAPDH foi de 33,04 ± 18,09%; enquanto que a média da proporção ETr-B/GAPDH foi de 114,58 ± 42,30%. Em relação às características histopatológicas individuais, a frequência de fibrose foi de 100%, infiltrado mononuclear de 88,23%, neovascularização de 52,94%, calcificação de 58,82% e houve ausência de ossificação. CONCLUSÃO: A presença de receptores ETr-A e ETr-B em valvas mitrais reumáticas sugere sua interação com o sistema de endotelinas circulantes, particularmente ETr-B (reconhecido por atuar na remoção do excesso de endotelina), detectado em maior proporção, o que poderia explicar a ausência da expressão de endotelina em valva mitral reumática, processo a ser elucidado.

Amplificação dos genes que codificam a endotelina-1 e seus receptores em valvas mitrais reumáticas / Amplification of the genes that codify endothelin-1 and its receptors in rheumatic mitral valves

FUNDAMENTO: As cardiopatias são doenças de alta prevalência, sendo a cardite reumática uma doença de grande relevância em países em desenvolvimento. As alterações em câmaras cardíacas esquerdas se associam à disfunção endotelial, com aumento dos níveis de endotelina-1 (ET-1) e consequências sobre a circulação pulmonar, muitas vezes determinando a hipertensão pulmonar (HP). No entanto, a presença de ET-1 e seus receptores na própria valva mitral, promovendo alterações vasculares pulmonares e aumentando a deformação valvar reumática, ainda é um assunto não abordado na literatura. OBJETIVO: Determinar, mediante técnicas moleculares, a expressão dos genes da endotelina e dos seus receptores em valvas mitrais reumáticas. MÉTODOS: 27 pacientes submetidos à troca valvar mitral tiveram seu tecido valvar analisado, a fim de determinar a presença de genes de ET-1 e seus receptores A e B. Foram feitas análises histológica e molecular das valvas (divididas em fragmentos M1, M2 e M3) e colhidos dados clínicos e epidemiológicos dos pacientes. Foram divididos em três grupos: valvopatia mitral, mitroaórtica e pacientes reoperados. RESULTADOS: O estudo mostrou a manifestação do gene da ET-1 em 40,7 por cento dos espécimes e de seu receptor A em todas as amostras, com manifestação minoritária do gene do receptor B (22,2 por cento). CONCLUSÃO: Todos os pacientes expressaram a presença do gene do receptor A. Não houve diferença estatística quanto à gravidade da doença, expressa em classe funcional, e aos subgrupos estudados (valvopatas mitrais, mitroaórticos e pacientes reoperados), ou quanto à expressão dos genes da ET-1 e seus receptores entre os subgrupos estudados (valvopatas mitrais, mitroaórticos e pacientes reoperados).

BACKGROUND: Cardiopathies are high prevalence conditions. Among them, rheumatic carditis is of high relevance in developing countries. Left cardiac chamber changes are associated to endothelial dysfunction and ET-1 levels increase. Pulmonary circulation is then affected, and not seldom leading to pulmonary hypertension (PH). However, the presence of ET-1 and its receptors in the mitral valve itself - promoting pulmonary vascular changes, with increased rheumatic valvular deformation - has not been discussed in the literature. OBJECTIVE: To determine the expression of endothelin gene and its receptors in rheumatic mitral valves through techniques of molecular genetics. METHODS: Twenty-seven patients submitted to mitral valve replacement had their valvular tissue examined to determine the presence of ET-1 genes and their A and B receptors. Histological and molecular analysis of the valves was performed (divided into M1, M2 and M3 fragments), with patients' clinical and epidemiological data collected. Patients were divided into 3 groups (mitral valvopathy, mitroaortic valvopathy, and reoperation patients). RESULTS: The study showed endothelin-1 gene expression in 40.7 percent specimens and A receptor in all samples; receptor gene B had lower expression (22.2 percent). CONCLUSION: All patients showed A receptor gene expression. No statistically significant difference was observed in regard to condition severity, expressed according to functional class, and subgroups (mitral valvopathy, mitroaortic valvopathy, and reoperation patients).

Association of tumor necrosis factor-alpha polymorphisms with susceptibility and clinical outcomes of rheumatic heart disease

To examine the association of tumor necrosis factor-alpha [TNF-alpha] gene polymorphisms with rheumatic heart disease [RHD] and valve damage, and their influence on TNF-alpha production and disease outcome. We performed this cross-sectional study at Kasr El-Aini Hospital, Cairo University, Cairo, Egypt, from December 2008 to October 2009. Eighty children with chronic RHD and valve affection, and 50 controls were included. Patients with any other diseases or complications were excluded. Blood samples [5 ml] were collected. Genotyping for TNF-alpha polymorphisms was performed by the polymerase chain reaction-restriction fragment length polymorphism method. Serum TNF-alpha was measured by enzyme-linked immunosorbent assay. Serum TNF-alpha was significantly increased in RHD compared with controls [p=0.00003]. The TNF-alpha-238 adenine [AA] [p=0.036] and-308AA [p=0.003] genotypes were more frequent in RHD patients than in controls, and were associated with increased production of TNF-alpha [p=0.00001 for 238AA] and [p=0.001 for 308AA]. Both polymorphisms contributed to increased susceptibility for RHD [-308AA and adenine guanine [AG], odds ratio [OR]=4.72 [95% confidence interval [CI] 2.03-11.05], p=0.0001]; [-238 AA and AG, OR=2.33 [CI: 1.05-5.19], p=0.035]. The presence of-308AA was associated with mitral [p=0.001] and multivalvular [p=0.003] lesions and was more prevalent in moderate [p=0.001], and severe [p<0.001] cases than in controls. The-238AA variant was associated with mitral lesions [p=0.04] and severe cases [p=0.05] as compared with controls. The TNF-alpha-238G/A and-308G/A polymorphisms were associated with susceptibility to RHD and increased production of TNF-alpha. Both polymorphisms were related to valve damage, and a more severe outcome of RHD

HLA Profile in Iraqi rheumatic valvulitis patients

Human leukocyte antigen [LILA] is the most polymorphic genetic system in man. The genes of this region influence susceptibility to certain disease. This study was established to shed light on the possible association of HLA class I and II antigens with RV patients. Lymphocvtotoxicity assay for HLA for class I and II typing had been done for [100] iraqi patients suffering from rheumatic valvulitis [RV] the control groups consisting of [75 healthy individuals and 35 non rheumatic heart disease [NRHD] patients]. The results showed a significant association of A33-Ag with these patients as compared with healthy and cardiac controls [P=0.005][P=0.033] respectively. Another interesting finding was the low frequency of AI in RI patients when compared with heal/he control [p=0.002], suggesting that Al allele may confer protective effect against this disease. In addition significant association between blood group B and RV was evident [p=0.04] An interesting observation was a strong association of blood group B and A33 among those patients [P < 0.001]. The present results are consistent with hypothesis that susceptibility to RV is genetically linked and in turn may be associated mainly with A33 in Iraqi patients

Níveis de citocinas proinflamatórias e seus antagonistas em pacientes com insuficiência aórtica crônica importante / Proninflamatory cytokine and antagonists levels with chronic severe aortic regurgitation

Realizamos estudo transversal com um grupo homogêneo constituído de 90 pacientes com insuficiência aórtica crônica importante, de etiologia reumática. Foram realizadas dosagens plasmáticas de citocinas proninfmamatórias a saber : Fator de necrose tumoral alfa( TNF ), receptores solúveis de TNF tipo I e II ( sTNFRI e sTNFRII ), interleucina 6 ( IL-6 ) e seu receptor solúvel ( IL-6R ), inteleucina 1-beta ( IL-1B ), antagonista do receptor de IL-1 ( IL1-RA ) e endotelina-1. Realizado o polimorfismo –308 do gene do TNF. Observamos níveis aumentados de TNF, IL-6 e sTNFRI em relação a indivíduos normais. Pacientes sintomáticos e assintomáticos tiveram níveis semelhantes de citocinas. O polimorfismo TNF 1/2 foi mais frequente em indivíduos assintomáticos. O aumento dos diâmetros ventriculares correlacionou-se a diminuição dos níveis de sTNFR II. /We made a transversal study in a homogeneous group of rheumatic patients with chronic severe aortic regurgitation. We determined plasma levels of the following proinflamatory cytokines : Tumor necrosis factor-alpha ( TNF ) and its soluble receptors type I and II ( sTNFR I and sTNFR II ), Interleukin-6 ( IL-6 ) and its soluble receptor ( IL-6R ), interleukin-1 beta ( IL-1 beta ), antagonist of the IL-1 receptor ( IL1-RA ) and endothelin-1. The -308 genetic polimorphism os the TNF gene was made. Plasma levels of TNF, IL-6 and sTNFRI were increased in asymptomatic and symptomatic in relation to controls. There were similar levels of cytokines in asymptromatic and symptomatic patients. The polimorphism TNF 1/2 was more frequent in asymptomatic patients. Increase of ventricular diameters was correlated to decrease...

HLA-DR types in rheumatic, congenital and degenerative cardiac valve diseasesin Saudi Patients

Subjects of Saudi origin were DNA typed for HLD-DR2, DR4 and DR[w]53 by fragment-length polymorphism and amplification by sequence-specific primer techniques based on polymerase chain reaction. Of these subjects, 125 sporadic heart valve disease [96 with rheumatic heart disease, 18 with degenerate and 11 with congenital degenerate valve disease] and 77 were healthy Saudi blood donors. While the frequency of individuals typed DR[4] was about the same in the rheumatic heart disease as in the control category [30% versus 23%, respectively], it was found to be higher [55%; P< 0.02], but below the level of marginal significance after correcting for the number of DR types, in the congenital degenerate valve category. No preferential association of any DR4 subtype could be detected. The incidence of DR2 was lower in the congenital cases compared to that in the controls [9% versus 21%] and remained about the same in the rheumatic heart disease patents as in the controls. The frequency of DR[w]53 in the degenerate valve categories was slightly lower than that in the controls, but the difference was not significant. The study failed to corroborate the association between HLA-DR4 and rheumatic heart disease shown in previous studies using the sterotyping approach