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肝脾血吸虫病患者水通道蛋白-2 和 NKCC2 的表达模式。

Aquaporin-2 and NKCC2 expression pattern in patients with hepatosplenic schistosomiasis.

机构信息

Post-Graduation Program in Medical Sciences, Department of Internal Medicine, Federal University of Ceará, Fortaleza, Brazil.

School of Medicine, Federal University of Alagoas, Maceió, Brazil.

出版信息

Trop Med Int Health. 2020 Sep;25(9):1140-1144. doi: 10.1111/tmi.13461. Epub 2020 Jul 30.

Abstract

OBJECTIVE

Schistosoma mansoni infection is considered a public health problem. Glomerular involvement in schistosomiasis is a well-documented complication, especially in hepatosplenic schistosomiasis (HSS). However, renal tubular function is poorly understood. The aim of this study was to investigate, through urinary exosomes, tubular transporters functionally in HSS patients.

METHODS

Cross-sectional study of 20 HSS patients who had isolated exosomes from urine samples. Protease inhibitor was added in the urine samples who were immediately frozen at -80 °C for further exosomes isolation. After urine had thawed, urinary exosomes were obtained using extensive vortexing, centrifugation and ultracentrifugation steps of urine. Urinary transporters expression from exosomes was evaluated by western blot, including NHE3, AQP2 and NKCC2. Charge amounts for gel electrophoresis were adjusted by urinary creatinine concentration of each patient to avoid urinary concentration bias. All protein expression of HSS patients was relative to healthy controls.

RESULTS

The expression of aquaporin-2 (AQP2) was lower in HSS patients than in controls (46.8 ± 40.7 vs. 100 ± 70.2%, P = 0.03) and the expression of the NKCC2 co-transporter was higher (191.7 ± 248.6 vs. 100 ± 43.6%, P = 0.02).

CONCLUSIONS

The decrease of AQP2 and the increase of NKCC2 expression in HSS patients seem to be involved with the inability of urinary concentration in these patients. These data show renal tubular abnormalities in HSS patients without manifest clinical disease.

摘要

目的

曼氏血吸虫感染被认为是一个公共卫生问题。肾小球受累是血吸虫病的一种有据可查的并发症,尤其是在肝脾血吸虫病(HSS)中。然而,肾小管功能仍知之甚少。本研究旨在通过尿外泌体研究 HSS 患者的肾小管转运体的功能。

方法

对 20 例 HSS 患者进行横断面研究,从尿液样本中分离出外泌体。在尿液中加入蛋白酶抑制剂,立即在-80°C 冷冻,以便进一步分离外泌体。尿液解冻后,通过广泛涡旋、离心和超速离心步骤获得尿液外泌体。通过 Western blot 评估外泌体中肾小管转运体的表达,包括 NHE3、AQP2 和 NKCC2。通过每个患者的尿液肌酐浓度调整凝胶电泳的电荷量,以避免尿液浓度偏差。所有 HSS 患者的蛋白表达均与健康对照组进行比较。

结果

HSS 患者的水通道蛋白-2(AQP2)表达低于对照组(46.8±40.7 对 100±70.2%,P=0.03),NKCC2 协同转运体的表达更高(191.7±248.6 对 100±43.6%,P=0.02)。

结论

HSS 患者 AQP2 表达降低和 NKCC2 表达增加似乎与这些患者的尿液浓缩能力丧失有关。这些数据显示 HSS 患者在没有明显临床疾病的情况下存在肾小管异常。

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