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American journal of physiology. Heart and circulatory physiology, 2020-02, Vol.318 (2), p.H448-H460
2020

Details

Autor(en) / Beteiligte
Titel
Several phosphate transport processes are present in vascular smooth muscle cells
Ist Teil von
  • American journal of physiology. Heart and circulatory physiology, 2020-02, Vol.318 (2), p.H448-H460
Ort / Verlag
United States: American Physiological Society
Erscheinungsjahr
2020
Link zum Volltext
Quelle
EZB Free E-Journals
Beschreibungen/Notizen
  • We have studied inorganic phosphate (P ) handling in rat aortic vascular smooth muscle cells (VSMC) using P-radiotracer assays. Our results have revealed a complex set of mechanisms consisting of ) well-known PiT1/PiT2-mediated sodium-dependent P transport; ) Slc20-unrelated sodium-dependent P transport that is sensitive to the stilbene derivatives 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) and 4-acetamido-4-isothiocyanostilbene-2,2-disulfonate (SITS); ) a sodium-independent P uptake system that is competitively inhibited by sulfate, bicarbonate, and arsenate and is weakly inhibited by DIDS, SITS, and phosphonoformate; and ) an exit pathway from the cell that is partially chloride dependent and unrelated to the known anion-exchangers expressed in VSMC. The inhibitions of sodium-independent P transport by sulfate and of sodium-dependent transport by SITS were studied in greater detail. The maximal inhibition by sulfate was similar to that of P itself, with a very high inhibition constant (212 mM). SITS only partially inhibited sodium-dependent P transport, but the was very low (14 µM). Nevertheless, SITS and DIDS did not inhibit P transport in oocytes expressing PiT1 or PiT2. Both the sodium-dependent and sodium-independent transport systems were highly dependent on VSMC confluence and on the differentiation state, but they were not modified by incubating VSMC for 7 days with 2 mM P under nonprecipitating conditions. This work not only shows that the P handling by cells is highly complex but also that the transport systems are shared with other ions such as bicarbonate or sulfate. In addition to the inorganic phosphate (P ) transporters PiT1 and PiT2, rat vascular smooth muscle cells show a sodium-dependent P transport system that is inhibited by DIDS and SITS. A sodium-independent P uptake system of high affinity is also expressed, which is inhibited by sulfate, bicarbonate, and arsenate. The exit of excess P is through an exchange with extracellular chloride. Whereas the metabolic effects of the inhibitors, if any, cannot be discarded, kinetic analysis during initial velocity suggests competitive inhibition.

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