- 學位論文
探討Dipyridamole 在 Anti-Thy-1抗體引發腎絲球腎炎動物模式中的抗發炎功能
The anti-inflammatory effects of dipyridamole in anti-Thy-1 antibody induced Glomerulonephritis
摘要
Anti-thy-1 抗體所引發的腎絲球腎炎，是一種常被引用來研究腎環門間質細胞( Mesangial cell )增生性腎絲球腎炎的動物模式。Dipyridamole是一種phosphodiesterase抑制劑，臨床上曾被用來治療由絲球體腎炎所引起的蛋白尿。然而，Dipyridamole於臨床上的治療機制目前尚未完全明瞭。由實驗室先前的研究顯示；發現到，施打anti-Thy-1抗體所引發的急性腎絲球腎炎，其症狀會出現明顯蛋白尿、ED(+)巨噬細胞移行、浸潤、及中度的PCNA(+)細胞增生。以上這些症狀經由Dipyridamole( 5mg/kg )治療的大鼠實驗可明顯觀察到抑制情形。TGF-β對於調控細胞外基質-collegen、fibronectin的分泌具重要的關鍵。我們也測試了Dipyridamole對於TGF-β表現和細胞間質增生、堆積的抑制，是否有相同的效果。由PAS score增加顯示；經由施打anti-Thy-1抗體所引發的少量腎絲球TGF-β增生和細胞基質堆積，以Dipyridamole治療後，發現在腎絲球PAS score和TGF-β表現有明顯的減少。 Heme-oxygenase-1(HO-1)是一種生理性壓力指標，Thy-1抗體所引起的HO-1表現會增加，在Dipyridamole治療後也會受到抑制。Osteopontin(OPN) 是一種bone sialoprotein，也曾被報導和腎臟纖維化有關。我們研究了OPN 在Thy-1抗體引發的腎炎所扮演的功能，發現OPN是一種組織的組成物質，於正常腎臟的腎小管、鮑氏囊體、及亨利氏環均有存在。施打anti-Thy-1抗體後於腎小管位置OPN有顯著的增加，但在絲球體卻沒有發現。所以，OPN不是和Thy-1產生的腎炎及腎纖維化有相關性。本實驗所使用Dipyridamole治療由Thy-1引起的OPN表現，於腎小管位置有發現受到抑制，但於腎小球沒有發現。綜合以上的報告，Dipyridamole 治療由anti-Thy-1引起的腎絲球腎炎蛋白尿，具有抑制的功效。
並列摘要
Anti-thy1 antibody-induced glomerular nephritis is a well established animal model for Mesangial cell proliferative glomeruli nephritis. Dipyridamole is a phosphodiesterase inhibitor, which has been shown to reduce GN-associated proteinurea. However, the detail mechanisms by which dipyridamole exert its therapeutic effects has not been elucidated. In the present study, we investigate the effect of dipyridamole on anti-thy1 -induced glomerular nephritis. We found that injection of anti-thy1 antibody resulted in acute glomerular nephritis (GN), characterized with massive proteinurea, increased of ED(+) macrophage infiltration and moderate PCNA(+) cell proliferation. These effects were all inhibited by treatment of rats with dipyridamole (5mg/Kg). TGF-β?浵as been shown to play an important role in extracellular matrix secretion. We next examined whether treatment of dipyridamole exerted beneficial effects on glomerular TGF-β expression and extracellular matrix accumulation. Injection of anti-thy1 antibody resulted in a mild increase of glomerular TGF-β and extracellular matrix accumulation as revealed by increased PAS score. Treatment of GN rats with dipyridamole significantly reduced the PAS score and TGF-β expresseion levels in glomeruli. Thy-1 antibody induced the expression of hem-oxygenase-1 (HO-1), a stress-induced marker, and the HO-1 induction was inhibited by treatment of rats with dipyridamole. Osteopontin, a bone sialoprotein, has been linked to renal fibrosis. I next investigate whether osteopontin (OPN) plays a role in anti Thy1 Ab-induced GN. OPN was constitutively expressed in tubules, Bowman’s capsule, and glomerular tuft of normal kidney. Injection of anti-thy1 antibody increased OPN expression in renal tubules but not in glomeruli, suggesting OPN was not associated with thy1-induced GN and subsequent renal fibrosis. Treatment of cells with dipyridamole reduced thy1-antibody stimulated OPN expression in renal tubule, but has little effect on glomerular OPN expression. Taken together, our results demonstrated that dipyridamole exhibited beneficial effects on thy-1 antibody induced GN.