Luan Haiyan, Wu Peng, Wang Mingxiao, Sui Hongyu, Fan Lili, Gu Ruimin
Department of Pharmacology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China.
Department of Physiology, Basic Medical School, Jiamusi University, Jiamusi, Heilongjiang 154007, P.R. China.
Mol Med Rep. 2016 Nov;14(5):4391-4398. doi: 10.3892/mmr.2016.5781. Epub 2016 Sep 26.
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.
腺苷是一种由体内多个器官产生的分子，包括肾脏，它在肾脏中作为一种自身调节因子发挥作用。它介导了几个肾单位节段中的离子转运，包括近端小管和髓袢升支粗段（TAL）。离子转运部分由阴离子氯通道决定，这些通道调节关键的肾脏功能，包括Na+和Cl‑的重吸收、尿液浓缩以及建立和维持皮质髓质渗透梯度。本研究调查了腺苷对氯离子电压门控通道Kb（CLCNKB）mRNA表达的影响，CLCNKB是一个与高血压相关的候选基因，编码ClC‑Kb通道。从大鼠肾脏分离出髓袢升支粗段（mTAL）小管，并建立了来自mTAL小管的mTAL细胞的原代培养物。用腺苷处理细胞，并通过逆转录定量聚合酶链反应检测CLCNKB的mRNA表达。还用通路抑制剂（H8和AACOCF3）处理细胞，并通过蛋白质印迹法分析环磷酸腺苷（cAMP）‑蛋白激酶A（PKA）和磷脂酶A2（PLA2）的蛋白表达。研究结果表明，腺苷增加了髓袢升支粗段细胞原代培养物中CLCNKB的mRNA表达，并且这种刺激作用受cAMP‑PKA和PLA2‑花生四烯酸（AA）通路调节。本研究表明，腺苷最初通过cAMP‑PKA通路，然后通过PLA2‑AA通路影响CLCNKB的mRNA表达。
