Fragilidade osmótica eritrocitária e reticulocitometria em cães acometidos por doença renal crônica
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Data
2020-03-13
Autores
Bizare, Amanda
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Universidade Estadual Paulista (Unesp)
Resumo
A anemia é considerada um dos fatores para avaliar progressão da doença renal e diminuição da qualidade de vida do paciente. Conforme a doença renal progride, ocorre aumento gradativo na produção de toxinas urêmicas que reduz a meia vida dos eritrócitos circulantes por interferir na estabilidade da membrana eritrocitária. Para tanto, utiliza-se a contagem de reticulócitos para classificar a anemia como regenerativa ou não regenerativa. Objetivou-se neste estudo avaliar a resistência da membrana das hemácias, utilizando-se do teste de Fragilidade Osmótica Eritrocitária (FOE) em cães com doença renal crônica (DRC) e avaliação de reticulócitos. Foram avaliados 43 cães provenientes da rotina do Serviço de Nefrologia e Urologia do Hospital Veterinário Governador Laudo Natel da Faculdade de Ciências Agrárias e Veterinárias - UNESP - campus Jaboticabal. As referidas unidades experimentais foram distribuídas em três grupos, quais sejam, G0 (n=13), composto por cães hígidos e G1, DRC estádios 1 e 2 (n=14) e G2, DRC estádios 3 e 4 (n=16), classificados de acordo com o recomendado pela International Renal Interest Society. A fim de definir os critérios de inclusão dos cães foram feitos, além do exame físico, a avaliação de pressão arterial, hemograma, contagem de reticulócitos, exames bioquímicos, urinálise e relação proteína/creatinina urinária (UP/C). Para execução do teste de FOE as hemácias foram diluídas em concentrações decrescentes de cloreto de sódio e analisadas por citometria de fluxo. As concentrações de creatinina sérica (sCr), ureia sérica (sUreia), fosfato sérico (sP) e UP/C foram superiores no grupo G2 quando comparado ao G0 (p<0,05). Com relação às variáveis hematológicas, foram observadas diferença estatística entre o número de hemácias (p<0,009), hematócrito (p<007) e hemoglobina (p<0,007), que decresceu conforme a doença renal avançava. A FOE e os reticulócitos apesar de terem variado individualmente, dependendo do estádio da DRC de cada paciente, não apresentaram diferença estatística quando comparado aos animais controle e doentes, e tampouco quando foram confrontados os diferentes estádios de cães com DRC (p>0,05). Os resultados obtidos neste estudo demonstram, portanto que, embora a reticulocitometria seja fiadora da contagem de reticulócitos, a contabilização manual de reticulócitos ainda é uma opção segura e somente com a FOE não é possível predizer alterações na membrana do eritrócito em cães com DRC.
Anemia is considered one of the factors to assess the kidney disease progress and the decrease in patient's quality of life. As kidney disease progresses, there is a gradual increase in urinary toxin production that shortens the circulating erythrocyte half-life by interfering with erythrocyte membrane stability. To do this, use a reticulocyte count to classify anemia as regenerative or non-regenerative. The objectives of this study were to evaluate the resistance of the red blood cells, using the Erythrocyte Osmotic Fragility test in dogs with chronic kidney disease (CKD) and to evaluate reticulocytes. Forty-three dogs were charged, followed by the routine of the Nephrology and Urology Service of the Governor Laudo Natel Veterinary Hospital of the Faculty of Agricultural and Veterinary Sciences - UNESP - Campus Jaboticabal. The experimental units were divided into three groups, namely, G0 (n = 13), consisting of healthy dogs and G1, CKD stages 1 and 2 (n = 14) and G2, CKD stages 3 and 4 (n = 16), classification proposed by the International Renal Interest Society. In order to define the inclusion criteria of dogs made, in addition to physical examination, an assessment of blood pressure, blood count, reticulocyte count, biochemical tests, urinalysis, and urinary protein/creatinine ratio. To perform the erythrocyte osmotic fragility test, red blood cells were diluted in decreasing sodium chloride filters (0.9 to 0.0%) and analyzed by flow cytometry. As creatinine serum concentration (sCr), serum urea (sUL), serum phosphorus (sP) and urinary protein/creatinine ratio. were higher in group G2 when compared to G0 (p <0.05). Regarding hematological variables, statistical differences were observed between the number of red blood cells (p <0.009), hematocrit (p <007) and hemoglobin (p <0.007), which decreased according to the advanced kidney disease. A cross-referenced FOE despite varying the number of participants, depending on each patient's CKD stage, shows no statistical difference when it shows control animals and damage, and shows when it is confronted with different cases of dogs with CKD (p> 0, 05) The results obtained in this study demonstrate, therefore, that although reticulocytometry is a guarantor of reticulocyte counting, manual reticulocyte counting is still a safe option and only with FOE it is not possible to predict changes in the erythrocyte membrane in dogs with CKD.
Anemia is considered one of the factors to assess the kidney disease progress and the decrease in patient's quality of life. As kidney disease progresses, there is a gradual increase in urinary toxin production that shortens the circulating erythrocyte half-life by interfering with erythrocyte membrane stability. To do this, use a reticulocyte count to classify anemia as regenerative or non-regenerative. The objectives of this study were to evaluate the resistance of the red blood cells, using the Erythrocyte Osmotic Fragility test in dogs with chronic kidney disease (CKD) and to evaluate reticulocytes. Forty-three dogs were charged, followed by the routine of the Nephrology and Urology Service of the Governor Laudo Natel Veterinary Hospital of the Faculty of Agricultural and Veterinary Sciences - UNESP - Campus Jaboticabal. The experimental units were divided into three groups, namely, G0 (n = 13), consisting of healthy dogs and G1, CKD stages 1 and 2 (n = 14) and G2, CKD stages 3 and 4 (n = 16), classification proposed by the International Renal Interest Society. In order to define the inclusion criteria of dogs made, in addition to physical examination, an assessment of blood pressure, blood count, reticulocyte count, biochemical tests, urinalysis, and urinary protein/creatinine ratio. To perform the erythrocyte osmotic fragility test, red blood cells were diluted in decreasing sodium chloride filters (0.9 to 0.0%) and analyzed by flow cytometry. As creatinine serum concentration (sCr), serum urea (sUL), serum phosphorus (sP) and urinary protein/creatinine ratio. were higher in group G2 when compared to G0 (p <0.05). Regarding hematological variables, statistical differences were observed between the number of red blood cells (p <0.009), hematocrit (p <007) and hemoglobin (p <0.007), which decreased according to the advanced kidney disease. A cross-referenced FOE despite varying the number of participants, depending on each patient's CKD stage, shows no statistical difference when it shows control animals and damage, and shows when it is confronted with different cases of dogs with CKD (p> 0, 05) The results obtained in this study demonstrate, therefore, that although reticulocytometry is a guarantor of reticulocyte counting, manual reticulocyte counting is still a safe option and only with FOE it is not possible to predict changes in the erythrocyte membrane in dogs with CKD.
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Palavras-chave
Citometria de fluxo, Hemácias, Hemólise, Nefropatia crônica, Flow citometry, Red blood cell, Hemolysis, Chronic nephropathy