Effects of adenosine stimulation on the mRNA expression of CLCNKB in the basolateral medullary thick ascending limb of the rat kidney.

Haiyan Luan, Peng Wu, Mingxiao Wang, Hongyu Sui, Lili Fan, Ruimin Gu

Molecular medicine reports 2016 Nov

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Adenosine is a molecule produced by several organs within the body, including the kidneys, where it acts as an autoregulatory factor. It mediates ion transport in several nephron segments, including the proximal tubule and the thick ascending limb (TAL). Ion transport is dictated in part by anionic chloride channels, which regulate crucial kidney functions, including the reabsorption of Na+ and Cl‑, urine concentration, and establishing and maintaining the corticomedullary osmotic gradient. The present study investigated the effects of adenosine on the mRNA expression of chloride voltage‑gated channel Kb (CLCNKB), a candidate gene involved in hypertension, which encodes for the ClC‑Kb channel. Medullary thick ascending limb (mTAL) tubules were isolated from the rat kidney, and primary cultures of mTAL cells from the mTAL tubules were established. The cells were treated with adenosine and the mRNA expression of CLCNKB was detected by reverse transcription‑quantitative polymerase chain reaction. The cells were also treated with pathways inhibitors (H8 and AACOCF3), and the protein expression of cyclic adenosine 3',5'‑monophosphate (cAMP)‑protein kinase A (PKA) and phospholipase A2 (PLA2) by were analyzed by western blotting. The findings indicated that adenosine increased the mRNA expression of CLCNKB in primary cultures of medullary TAL cells, and this stimulatory effect was regulated by the cAMP‑PKA and PLA2‑arachidonic acid (AA) pathways. The present study showed that adenosine affected the mRNA expression of CLCNKB, initially through the cAMP‑PKA pathway and then the PLA2‑AA pathway.

Citation

Haiyan Luan, Peng Wu, Mingxiao Wang, Hongyu Sui, Lili Fan, Ruimin Gu. Effects of adenosine stimulation on the mRNA expression of CLCNKB in the basolateral medullary thick ascending limb of the rat kidney. Molecular medicine reports. 2016 Nov;14(5):4391-4398