P2X4 receptor mediates interleukin-1β secretion of renal tubular epithelial cells under high glucose stimulation

doi:10.3877/cma.j.issn.2095-3216.2012.02.010

Abstract

Abstract:

Objective

To investigate the role of P2X4 in interleukin (IL)-1β secretion of renal tubular epithelial cell under high glucose stimulation.

Methods

Human proximal tubular epithelial cell line (HK-2) cells were grown in media with normal glucose concentration (5 mmol/L), high glucose concentration (15, 25, 35 and 50 mmol/L). HK-2 cells were incubated in media with high glucose(35 mmol/L, 48 h) with and without apyrase. HK-2 cells were transfected with P2X4 siRNA. Intracellular K⁺ concentration was measured by PBFI-AM. Cytosolic free Ca²⁺ was indicated by Fluo-3/AM. Data from experiments were analyzed by t-test or ANOVA wherever appropriate.

Results

The IL-1β and adenosine triphosphate (ATP) levels of cell supernatants increased in a dose and time dependent manner under high glucose stimulation. Apyrase significantly inhibited IL-1β level elevations caused by high glucose (8.2±4.9 μg/L vs. 18.5±4.0 μg/L, P ＜ 0.05). High glucose decreased intracellular K⁺ and increased intracellular Ca²⁺ in HK-2 cell (P ＜ 0.05). The results suggested that high glucose activated P2X receptors of HK-2 cells. Silencing P2X4 with siRNA diminished the high glucose-induced expression of IL-1β (12.5±2.9 μg/L vs.20.3±5.3 μg/L，P ＜ 0.05).

Conclusion

These results demonstrate that the P2X4 mediates the secretion of IL-1β in renal tubular epithelial cells, which may promote the renal interstitial inflammation in diabetic nephropathy

Key words: P2X4, IL-1β, High glucose, Renal tubular epithelial cell

Ke-hong CHEN, Wei-wei ZHANG, Jin-hua ZHANG, Ju-rong YANG, Kai-long LI, Jian-guo ZHANG, Ya-ni HE. P2X4 receptor mediates interleukin-1β secretion of renal tubular epithelial cells under high glucose stimulation[J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2012, 01(02): 109-115.

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Figures/Tables 7

References 29

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