1. Acid loading stimulates rat glomerular mesangial cells proliferation through Na+–H+ exchanger isoform 1 (NHE1)-dependent pathway
- Author
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Ming-min Zhang, Wei Su, Jakub Fichna, Lin-yun Lai, Yong Gu, Shao-jun Liu, Shanyan Lin, Man Li, Chuan-Ming Hao, Kun Li, Yong-Yu Li, Chang-Jie Chen, and Ai Peng
- Subjects
medicine.medical_specialty ,Sodium-Hydrogen Exchangers ,Glomerular Mesangial Cell ,Blotting, Western ,Tetrazolium Salts ,Flow cytometry ,Amiloride ,Western blot ,Internal medicine ,medicine ,Animals ,MTT assay ,Renal Insufficiency, Chronic ,Cell Proliferation ,Pharmacology ,Sodium-Hydrogen Exchanger 1 ,Staining and Labeling ,medicine.diagnostic_test ,Chemistry ,Cell growth ,Cell Cycle ,General Medicine ,Hydrogen-Ion Concentration ,Cell cycle ,Flow Cytometry ,Fluoresceins ,Molecular biology ,Rats ,Blot ,Thiazoles ,Sodium–hydrogen antiporter ,Endocrinology ,Mesangial Cells ,Disease Progression ,Acidosis - Abstract
The role of metabolic acidosis in the progression of chronic kidney disease (CKD) remains unclear. The aim of the present study was to investigate the direct effects of acid loading on the proliferation of rat glomerular mesangial cells (GMCs) in vitro and the possible role of sodium-hydrogen ion exchanger isoform 1 (NHE1). Rat GMCs were treated with acidic medium as acid loading. Growth and proliferation of GMCs was studied by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, thymidine ((3)H-TdR) incorporation, and flow cytometry. NHE1 protein expression and activity were quantified by Western blot and dual wavelength epifluorescent illumination with 2',7'-bis-(2-carboxyethyl)-5(6)-carboxyfluorescein, respectively. 5-(N,N-dimethyl) amiloride hydrochloride (DMA), a specific inhibitor of NHE1, was used to investigate the possible involvement of NHE1 in the proliferation of GMCs. The MTT assay, (3)H-TdR incorporation, and cell cycle distribution analysis indicated that acid loading stimulated the proliferation of GMCs. Acid loading increased NHE1 activity, but had no effects on NHE1 expression at the protein level. The effects of acid loading on the proliferation of GMCs were inhibited by DMA. Acid loading induced GMC proliferation through NHE1-dependent pathways. Our findings may contribute to the understanding of metabolic acidosis in the progression of CKD.
- Published
- 2013