朱宗信2018-07-122018-07-122003http://ntur.lib.ntu.edu.tw//handle/246246/22627副甲狀腺素（parathyroid hormone）抑制近端腎小管對磷、鈉、重碳酸根的再吸收，近端腎小管經由鈉氫交換器再吸收鈉及重碳酸根，而近端腎小管腔側膜鈉氫交換器為NHE-3，負鼠腎（OKP）細胞是腎小管細胞，可表現NHE-3。目前已知有多種因子可調節NHE-3，如酸可增加NHE-3 活性、NHE-3 mRNA 表達及NHE-3 蛋白質表現量。在腎小管細胞急性給予副甲狀腺素可抑制蛋白質激脢A 及C 而抑制NHE-3。本計畫的研究目的是探討慢性給予副甲狀腺素對腎小管NHE-3 活性、NHE-3 蛋白質量的調節。 我們的方法是以負鼠腎（OKP）細胞為研究模式。細胞內pH 測定利用螢光物質BCECF 在不同pH 值下，其螢光強度不同，而以光譜螢光儀來測定，鈉氫交換器活性的測定乃給予細胞酸負荷後，再加鈉後看其pH 恢復速率而求得。以各種濃度之副甲狀腺素慢性給予後觀察對腎小管鈉氫交換器活性的作用。此外，亦以西方轉漬法來看副甲狀腺素對腎小管細胞NHE-3 蛋白質表現量的調節。 結果發現在OKP 細胞給予副甲狀腺素後其鈉氫交換器活性並無顯著變化。同樣地，在OKP 細胞給予副甲狀腺素後其NHE3 蛋白質表現量也無顯著變化。由此推論副甲狀腺素的作用主要在鈣、磷，而對鈉及酸的調節可能較無作用。Parathyroid hormone（PTH）decreases phosphate, sodium and bicarbonate reabsorption by the proximal tubule. Proximal tubule transcellular NaCl and NaHCO3 reabsorption is mediated by an apical membrane Na/H exchanger,（NHE-3）and a basolateral Na/HCO3（NBC-1）.OKP cells, a cell line of renal proximal tubule, express NHE-3 . The regulating factors of NHE-3 are multiple. Acid incubation increases NHE-3 activity, NHE-3 mRNA abundance, NHE-3 protein abundance. PTH acutely inhibits NHE-3 in OKP cells via a dual signaling cascade involving protein kinase A and C. The purpose of this project is to study the chronic effect of PTH on NHE-3 activity, NHE-3 protein abundance. We used OKP cell as the experiment model. Continuous measurement of intracellular pH was accomplished in cells using pH-sensitive dye BCECF. Na/H exchanger activity was assayed as the initial rate of Na-dependent pHi recovery from an acid load. We checked chronic effect of PTH in different concentration on Na/H exchange activity in renal tubular cell. We checked the effect of PTH on NHE-3 protein abundance in OKP cell. The results showed that there was no significant change of NHE-3 activity or NHE-3 protein abundance by PTH in OKP cells. We think the major effect of PTH is for Ca & P, not for Na & acid.application/pdf28262 bytesapplication/pdfzh-TW國立臺灣大學醫學院一般醫學科parathyroid hormonerenal tubuleNa/H exchanger副甲狀腺素腎小管鈉氫交換器journal articlehttp://ntur.lib.ntu.edu.tw/bitstream/246246/22627/1/912314B002350.pdf