Impaired Na+ -K+ -ATPase signaling in renal proximal tubule contributes to hyperuricemia-induced renal tubular injury

Experimental & Molecular Medicine
Jing XiaoZhibin Ye

Abstract

Hyperuricemia contributes to renal inflammation. We aimed to investigate the role of Na+-K+-ATPase (NKA) in hyperuricemia-induced renal tubular injury. Human primary proximal tubular epithelial cells (PTECs) were incubated with uric acid (UA) at increasing doses or for increasing lengths of time. PTECs were then stimulated by pre-incubation with an NKA α1 expression vector or small interfering RNA before UA (100 μg ml-1, 48 h) stimulation. Hyperuricemic rats were induced by gastric oxonic acid and treated with febuxostat (Feb). ATP levels, the activity of NKA and expression of its α1 subunit, Src, NOD-like receptor pyrin domain-containing protein 3 (NLRP3) and interleukin 1β (IL-1β) were measured both in vitro and in vivo. Beginning at concentrations of 100 μg ml-1, UA started to dose-dependently reduce NKA activity. UA at a concentration of 100 μg ml-1 time-dependently affected the NKA activity, with the maximal increased NKA activity at 24 h, but the activity started to decrease after 48 h. This inhibitory effect of UA on NKA activity at 48 h was in addition to a decrease in NKA α1 expression in the cell membrane, but an increase in lysosomes. This process also involved the subsequent activation of Src kinase and NLRP3, promo...Continue Reading

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Citations

Jul 25, 2018·International Journal of Molecular Sciences·Xiaoming FanJiang Tian
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Methods Mentioned

BETA
Flow Cytometry
urine collection
light microscopy
enzyme-linked immunosorbent assay
Assay
transfection
protein assay

Software Mentioned

SPSS

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