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Previous issue date: 2013 / Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / A variabilidade clínica descrita na doença falciforme (DF) tem sido associada ao efeito
epistático de vários genes, a exemplo do gene Klotho (Kl), cujos polimorfismos interferem na
regulação de canais de potássio (K+), cálcio (Ca2+) e fósforo, na expressão de vitamina D
(VitD) e paratormônio (PTH), e na supressão do estresse oxidativo. Com isto, o presente
estudo teve por objetivos investigar a frequência de SNPs no Kl em indivíduos com
hemoglobinopatia SC (HbSC), associando-os a marcadores de gravidade e subfenótipos da
doença e investigar associações entre os níveis de K+, Ca2+, fósforo, VitD e PTH com tais
marcadores de gravidade. Foi desenvolvido um estudo de corte transversal com 113
indivíduos com HbSC provenientes da Fundação HEMOBA, Salvador-Ba. As análises
hematológicas foram realizadas em contador eletrônico de células; o perfil de hemoglobinas
foi confirmado pela cromatografia líquida de alto desempenho; os marcadores lipídicos, de
hemólise, de função hepática e renal, Ca2+ e fósforo foram avaliados por método
colorimétrico, assim como o K+, por eletrodo de íon seletivo; as concentrações de VitD e PTH
foram investigadas por quimioluminescência e de anti-estreptolisina-O (ASLO) e proteína C
reativa (PCRe) por nefelometria; os SNPs no Kl (rs1207568, rs9527025, rs564481 e
rs648202) foram genotipados pelo ensaio de discriminação alélica pelo sistema TaqMan; os
haplótipos dos genes da globina beta foram investigados pela reação da polimerase em cadeia
- com restrição dos fragmentos com endonucleases de restrição (PCR-RFLP). Os dados
clínicos foram coletados em prontuários médicos. O SNP rs1207568 foi associado à
ocorrência de infecções (P=0,0170) e a níveis séricos elevados de albumina (P=0,0370), o
SNP rs648202 foi associado ao uso de medicações (P=0,0208) e o SNP rs9527025 a níveis
elevados de fósforo e bilirrubina direta (BD) (P=0,0044 e P=0,0092, respectivamente). O K+
foi positivamente correlacionado com os leucócitos (r=0,2916, P=0,0034), linfócitos típicos
(r=0,2644, P=0,0082), monócitos (r=0,2370, P=0,0182), plaquetas (r=0,4889, P<0,0001),
colesterol total (r=0,2521, P=0,0118), colesterol LDL (Col-LDL) (r=0,2953, P=0,0030),
fósforo (r=0,2447, P=0,0277), proteínas totais (PTs) (r=0,2415, P=0,0160), ferritina
(r=0,2263, P=0,0283), PCRe (r=0,2369, P=0,0222) e HbS (r=0,2474, P=0,0135). O K+ teve
correlação negativa com hemácias (r=-0,2076, P=0,0392) e Hb fetal (r=-0,2328, P=0,0204).
O Ca2+ apresentou correlação positiva com PTs (r=0,2991, P=0,0028) e albumina (r=0,3242,
P=0,0011) e negativa com o ASLO (r=-0,2216, P=0,0309). O fósforo foi negativamente
correlacionado com Hb (r=-0,3083, P=0,0051), hematócrito (r=-0,2610, P=0,0186),
bilirrubina indireta (r=-0,2685, P=0,0154) e creatinina (r=-0,3844, P=0,0004). Houve
correlação positiva entre fósforo e eosinófilos (r=0,4383, P<0,0001), linfócitos típicos
(r=0,2560, P=0,0211), plaquetas (r=0,3598, P=0,0010), as transaminases AST (r=0,3088,
P=0,0050) e ALT (r=0,2203, P=0,0481) e com a fosfatase alcalina (r=0,5185, P<0,0001). A
VitD foi correlacionada negativamente com ferro (r=-0,3459, P=0,0006), volume corpuscular
médio (r=-0,2071, P=0,0441), BD (r=-0,2629, P=0,0101) e PTH (r=-0,3689, P=0,0004), e
positivamente com reticulócitos (r=0,2361, P=0,0220), linfócitos típicos (r=0,2306,
P=0,0245) e PCRe (r=0,2113, P=0,0420). O PTH teve correlação negativa com leucócitos
(r=-0,2618, P=0,0143), linfócitos típicos (r=-0,2966, P=0,0053), monócitos (r=-0,2723,
P=0,0107), colesterol total (r=-0,2282, P=0,0335), Col-LDL (r=-0,2470, P=0,0211), BD (r=-
0,2462, P=0,0443) e AST (r=-0,2274, P=0,0342). Neste estudo, o K+, fósforo, VitD, PTH e
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os SNPs no Kl se mostraram úteis para a avaliação da gravidade clínica em indivíduos com HbSC. / The clinical variability described in sickle cell disease (SCD) has been linked to epistatic
effect of various genes, such as Klotho (Kl), whose polymorphisms affect the potassium (K+),
calcium (Ca2+) and phosphorus channels regulation, the vitamin D (VitD) and parathyroid
hormone (PTH) expression, and oxidative stress suppression. With this, the present study
aims to investigate the Kl SNPs frequency in hemoglobinopathy SC (HbSC) individuals,
associating them with markers of severity and disease sub-phenotypes, and to investigate
associations between K+, Ca2+, phosphorus, VitD and PTH levels with such gravity markers.
We developed a cross-sectional study of 113 individuals with HbSC from HEMOBA
Foundation, Salvador - Ba. The hematological values were determined in electronic cell
counter; the hemoglobin (Hb) profile were confirmed by high performance liquid
chromatography; the lipid, hemolysis, liver and kidney markers, Ca2+ and phosphorus were
evaluated by colorimetric method, as well as the K+ by ion selective electrode; the VitD and
PTH concentrations were investigated by chemiluminescence and the anti-streptolysin O
(ASO) and C-reactive protein (PCRe) by nephelometry; the Kl SNPs (rs1207568, rs9527025,
rs564481 and rs648202) were genotyped by allelic discrimination assay by TaqMan system
for genotyping; the beta globin genes haplotypes were investigated by polymerase chain
reaction - restriction fragment length polimorphism (PCR-RFLP). Clinical data were collected
from medical records. The SNP rs1207568 was associated with the infection occurrence
(P=0.0170) and raised serum albumin levels (P=0.0370), the SNP rs648202 was associated
with the medications use (P=0.0208) and the SNP rs9527025 with raised direct bilirubin (BD)
and phosphorus levels (P=0.0044 and P=0.0092, respectively). The K+ was positively
correlated with leukocytes (r=0.2916, P=0.0034), typical lymphocytes (r=0.2644, P=0.0082),
monocytes (r=0.2370, P=0,0182), platelets (r=0.4889, P<0.0001), total cholesterol (r=0.2521,
P=0.0118), LDL cholesterol (Col-LDL) (r=0.2953, P=0.0030), phosphorus (r=0.2447,
P=0.0277), total proteins (PTs) (r=0.2415, P=0.0160), ferritin (r=0.2263, P=0.0283), PCRe
(r=0.2369, P=0.0222) and Hb (r=0.2474, P=0.0135). The K+ had a negative correlation with
red blood cells (r=-0.2076, P=0.0392) and fetal Hb (r=-0.2328, P=0.0204). The Ca2+ was
positively correlated with PTs (r=0.2991, P=0.0028) and albumin (r=0.3242, P=0.0011) and
negatively with the ASO (r=-0.2216, P=0,0309). The phosphorus was negatively correlated
with Hb (r=-0.3083, P=0.0051), hematocrit (r=-0.2610, P=0.0186), indirect bilirubin (r=-
0.2685, P=0.0154) and creatinine (r=-0.3844, P=0.0004). There was a positive correlation
between phosphorus and eosinophils (r=0.4383, P<0.0001), typical lymphocytes (r=0.2560,
P=0.0211), platelet count (r=0.3598, P=0.0010), the transaminases AST (r=0.3088,
P=0.0050) and ALT (r=0.2203, P=0.0481) and with the alkaline phosphatase (r=0.5185,
P<0.0001). The VitD was negatively correlated with iron (r=-0.3459, P=0.0006), mean
corpuscular volume (r=-0.2071, P=0.0441), BD (r=-0.2629, P=0.0101) and PTH (r=-0.3689,
P=0.0004), and positively with reticulocytes (r=0.2361, P=0.0220), typical lymphocytes
(r=0.2306, P=0.0245) and PCRe (r=0.2113, P=0.0420). The PTH had a negative correlation
with leukocytes (r=-0.2618, P=0.0143), typical lymphocytes (r=-0.2966, P=0.0053),
monocytes (r=-0.2723, P=0.0107) and total cholesterol (r=-0.2282, P=0.0335), Col-LDL (r=-
0.2470, P=0.0211), BD (r=-0.2462, P=0.0443) and AST (r=-0.2274, P=0.0342). In this study,
10
the K+, phosphorus, VitD, PTH and the Kl SNPs evaluated proved useful parameter for
evaluating the clinical severity in individuals with HbSC.
| Identifer | oai:union.ndltd.org:IBICT/oai:www.arca.fiocruz.br:icict/7358 |
| Date | January 2013 |
| Creators | Pacheco, Ana Paula Almeida de Souza |
| Contributors | Figueiredo, Maria Stella, Acosta, Angelina Xavier, Gonçalves, Marilda de Souza, Gonçalves, Marilda de Souza |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | Portuguese |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da FIOCRUZ, instname:Fundação Oswaldo Cruz, instacron:FIOCRUZ |
| Rights | info:eu-repo/semantics/openAccess |
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