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題 名 | Functional Characterization of Intracellular pH Regulators Responsible for Acid Extrusion in Human Radial Artery Smooth Muscle Cells |
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作 者 | Lee, Chung-yi; Tsai, Yi-ting; Chang, Chung-yi; Chang, Yi-yu; Cheng, Tzu-hurng; Tsai, Chien-sung; Loh, Shih-hurng; | 書刊名 | The Chinese Journal of Physiology |
卷 期 | 57:5 2014.10[民103.10] |
頁 次 | 頁238-248 |
分類號 | 415.132 |
關鍵詞 | BCECF; Human radial artery smooth muscle cells; HRASMCs; Intracellular pH; pHᵢ; Monocarboxylate transporters; MCT; Na⁺-H⁺ exchanger; NHE; Na⁺-HCO₃⁻ cotransporter; NBC; |
語 文 | 英文(English) |
英文摘要 | Intracellular pH (pHi ) is a critical factor influencing many important cellular functions. Acid extrusion carriers such as Na+ /H+ exchanger (NHE), Na+ /HCO3 - cotransporter (NBC) and monocarboxylate transporters (MCT) can be activated when cells are in an acidic condition (pHi < 7.1). Human radial artery smooth muscle cells (HRASMCs) are an important conduit in coronary artery bypass graft surgery. However, such far, the pHi regulators have not been characterized in HRASMCs. We therefore investigated the mechanism of pHi recovery from intracellular acidosis and alkalosis, induced by NH4Clprepulse and Na-acetate-prepulse, respectively, using intracellular 2’,7’-bis(2-carboxethyl)-5(6)- carboxy-fluorescein (BCECF)-fluorescence in HRASMCs. Cultured HRASMCs were derived from the segments of human radial artery that were obtained from patients undergoing bypass grafting. The resting pHi is 7.22 ± 0.03 and 7.17 ± 0.02 for HEPES- (nominally HCO3 - -free) and CO2/HCO3 - - buffered solution, respectively. In HEPES-buffered solution, a pHi recovery from induced intracellular acidosis could be blocked completely by 30 µM HOE 694 (3-methylsulfonyl-4-piperidinobenzoyl, guanidine hydrochloride) a specific NHE inhibitor, or by removing [Na+ ]o. In 3% CO2/HCO3 - -buffered solution, HOE 694 slowed the pHi recovery from the induced intracellular acidosis only, while adding together with DIDS (a specific NBC inhibitor) or removal of [Na+ ]o entirely inhibited the acid extrusion. Moreover, α-cyano-4-hydroxycinnamate (CHC; a specific blocker of MCT) blocked the lactate-induced pHi changes. In conclusion, we demonstrate, for the first time, those 3 different pHi regulators responsible for acid extruding, i.e. NHE and NBC, and MCT, are functionally co-existed in cultured HRASMCs. |
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