Gerbino Chiara, Foglietta Federica, Corsi Daniele, Nardini Patrizia, Cangemi Luigi, Benetti Elisa, Rosa Arianna Carolina
Department of Scienza E Tecnologia del Farmaco, University of Turin, Via P. Giuria 9, 10125, Turin, Italy.
Iclas - GVM Care & Research, Via Mario Puchoz, 25, 16035, Rapallo, Italy.
Inflamm Res. 2025 Sep 12;74(1):122. doi: 10.1007/s00011-025-02095-4.
Investigate the potential role of histamine and its receptors on the functional expression of the sodium/hydrogen (Na/H) exchanger (NHE)3.
The human epithelial kidney (HK-2) cells were used as an in vitro model of the renal proximal tubule.
HK-2 cells were exposed to histamine 0-1000 nM alone or in combination with chlorphenamine (10 μM) and JNJ-7777120 (1 μM) for 0-48 h. MAPK involvement was determined using the selective inhibitors SB202190 (p38 MAPK), PD98059 (ERK1/2), and SP600125 (SAPK/JNK).
Gene and protein expression were evaluated by qPCR and immunoblotting. The activity of NHE3 was measured by the BCECF-AM-based method.
Histamine (100 nM) induced a concentration-dependent NHE3 gene transcription with a peak 16 h after the treatment, followed by protein translation at 48 h after. A Consistent increase in NHE3 activity was observed at 48 h, but also at 60 min, when both p38 MAPK and ERK1/2 were phosphorylated. JNJ-7777120 blunted the activation and expression of NHE3. Chlorpheniramine was effective only on NHE3 activity.
Histamine shows early (within 60 min) and late (48 h) effects on NHE3 expression. The histamine H and H receptors are shown to contribute to these effects differentially. The findings of this study extends the evidence for a direct contribution of histamine on the renal reabsorptive machinery.
研究组胺及其受体对钠/氢（Na/H）交换体（NHE）3功能表达的潜在作用。
人肾上皮细胞（HK-2）用作近端肾小管的体外模型。
HK-2细胞单独暴露于0 - 1000 nM组胺，或与氯苯那敏（10 μM）和JNJ-7777120（1 μM）联合处理0 - 48小时。使用选择性抑制剂SB202190（p38丝裂原活化蛋白激酶）、PD98059（细胞外信号调节激酶1/2）和SP600125（应激激活蛋白激酶/应激活化蛋白激酶）确定丝裂原活化蛋白激酶的参与情况。
通过定量聚合酶链反应和免疫印迹评估基因和蛋白质表达。采用基于2',7'-双（2-羧乙基）-5（6）-羧基荧光素乙酰甲酯（BCECF-AM）的方法测量NHE3的活性。
组胺（100 nM）诱导NHE3基因转录呈浓度依赖性，处理后16小时达到峰值，随后在48小时进行蛋白质翻译。在48小时以及60分钟时观察到NHE3活性持续增加，此时p38丝裂原活化蛋白激酶和细胞外信号调节激酶1/2均被磷酸化。JNJ-7777120抑制了NHE3的激活和表达。氯苯那敏仅对NHE3活性有效。
组胺对NHE3表达显示出早期（60分钟内）和晚期（48小时）效应。组胺H1和H2受体对这些效应的贡献有所不同。本研究结果扩展了组胺对肾脏重吸收机制有直接作用的证据。
