Dipyridamole 透过血基质氧化酵素-1抑制骨桥素表现.pptVIP

Dipyridamole透過血基質氧化酵素-1抑制骨橋素表現 骨橋素 osteopontin; OPN 是一種分泌型的磷酸化醣蛋白，在缺血性腎臟間質纖維化的過程裡扮演重要的角色。在NRK-52E大鼠腎小管上皮細胞加入化學缺氧劑氯化鈷 cobalt chloride; CoCl2 ，會誘導時間與劑量依存性OPN蛋白質表現增加。以l-N-acetylcysteine l-NAC 和thenoyl-trifluoroacetone TTFA 前處理後可阻斷CoCl2誘導OPN的增加，顯示自由基和電子傳遞複合物II可能是有關聯性的。以dipyridamole 1-10?M 前處理NRK-52E細胞，可抑制CoCl2誘導OPN的表現。因為誘導血基質氧化酵素-1 heme oxygenase-1; HO-1 的藥物已知可改善腎臟纖維化，而OPN的表現與腎臟間質纖維化有關，我們接著探討誘導HO-1的表現是否能調控OPN的表現。許多類固醇及非類固醇抗發炎藥劑均可以抑制由CoCl2所誘導的OPN表現。這些抗發炎藥劑中，rosiglitazone和dexamethasone無法誘導HO-1的表現，而celebrex和tanshinones IIA 能夠誘導HO-1的表現。以SnPP HO-1抑制劑 前處理NRK-52E 細胞，可逆轉dipyridamole的抑制作用，顯示HO-1與dipyridamole抑制CoCl2誘導OPN的表現有關。培養細胞時加入CO釋放分子 tricarbonyldichloro-ruthenium II ，能模擬dipyridamole抑制OPN表現的作用。以CO清除劑 血紅素；hemoglobin 前處理則能逆轉dipyridamole的抑制作用。這些資料顯示dipyridamole可能是透過誘導HO-1表現而抑制OPN的表現。增加HO-1可以催化血基質 heme 轉化出CO，進而抑制CoCl2誘導OPN的表現。我們的實驗結果顯示，HO-1的誘發劑可能適合做為腎臟纖維化的治療藥物。 Inhibition of Osteopontin Expression in NRK-52E Cells by Dipyridamole Is Mediated Through Heme Oxygenase-1 Osteopontin OPN , a secreted phospho-glycoprotein, plays a pivotal role in the progression of interstitial fibrosis in renal ischemia. In the present study, we showed that incubation of rat renal tubular NRK-52E cells with cobalt chloride CoCl2 increased OPN protein expression in a dose- and time- dependent manner. CoCl2 induced OPN expression was blocked by pretreatment with l-N-acetylcysteine l-NAC and thenoyl-trifluoroacetone TTFA suggesting free radical production and electron transport complex II were involved. Pretreatment of cells with dipyridamole 1-10 ?M , inhibited the CoCl2-induced OPN expression and the inhibitory effect was associated with heme oxygenase-1 HO-1 induction. Because induction of heme oxygenase-1 has been linked to inhibition of renal fibrosis, we tested whether induction of HO-1 modulate OPN expression in NRK-52E cells. In addition, we demonstrated that several agents that exert anti-inflammatory effects inhibited CoCl2-induced OPN expression. Among these anti-inflammatory