第一型胞漿素原活化抑制劑與環孢靈誘發牙齦增生關係之探討

環孢靈(cyclosporin A, 簡稱CsA)是臨床上被廣泛使用的免疫抑制劑，使用環孢靈後，大約百分之三十的病人會出現牙齦增生(gingival overgrowth)的副作用，而造成牙齦增生的主要原因是牙齦細胞外基質(extracellular matrix, 簡稱ECM)產生異常堆積的結果。第一型胞漿素原活化抑制劑(plasminogen activator inhibitor-1, 簡稱PAI-1)可以抑制ECM被分解，並與一些纖維化(fibrosis)的疾病有著密切的關係，然而目前有關PAI-1與CsA誘發牙齦增生的關係並不清楚。因此，本研究首先利用免疫組織化學染色法(immunohistochemistry)，探討正常牙齦與CsA誘發增生牙齦中PAI-1的表達；結果發現PAI-1表達在結締組織的發炎細胞和纖維母細胞上，並且在增生牙齦的切片中，可以觀察到較多的PAI-1表達。另培養人類正常牙齦纖維母細胞(human gingival fibroblast, 簡稱HGF)，利用反轉錄聚合酶連鎖反應(reverse transcriptase-polymerase chain reaction)、西方點墨法(Western blotting)及酵素連接免疫吸附分析法(enzyme-linked immunosorbent assay)，探討HGF加入CsA後，PAI-1是否會被誘發表現；另外，本研究再多加入牙周致病菌、發炎激素(proinflammatory cytokines)、N-acetylcysteine (簡稱NAC)、U0126和PD098059，探討CsA影響PAI-1表現的可能機轉。結果顯示，HGF加入CsA後，不論在messenger RNA或蛋白質的表現層次上，PAI-1的表現都是增加的(p<0.05)；另一方面，與單純加入CsA的結果相較，再多加入牙周致病菌或發炎激素，對於PAI-1的表現產生了上升的趨勢(p<0.05)；加入NAC、U0126和PD098059，PAI-1則明顯被抑制(p<0.05)。綜合研究結果顯示，CsA誘發PAI-1的表達可能是造成牙齦增生的原因之一，在牙周致病菌及發炎激素的刺激下，PAI-1的表現會更明顯，此外，CsA影響PAI-1表現的調控機轉，可能與細胞內glutathione及ERK-MAPK pathway有關。

關鍵字

環孢靈；牙齦增生；第一型胞漿素原活化抑制劑；調控機制

並列摘要

Cyclosporin A (CsA) is used as an immunosuppressive agent and its prominent side effect is the induction of fibrous gingival overgrowth. The progression of fibrous gingival overgrowth results from the accumulation of extracellular matrix (ECM). Plasminogen activator inhibitor-1 (PAI-1) acts as the inhibitor of ECM degradation, and involves with some fibrotic diseases. However, the correlation between PAI-1 and CsA-induced gingival overgrowth was little known, and it was the main purpose we investigated in this study. Immunochemistry was used to compare PAI-1 expression between gingival tissues obtained from normal patients and CsA-treated patients in vivo. The evaluations displayed that greater expression of PAI-1 was observed on fibroblasts and inflammatory cells in CsA-induced hyperplasia gingive. Reverse transcriptase-polymerase chain reaction、Western blotting and enzyme-linked immunosorbent assay were used to determine the effects of CsA on the expression of PAI-1 in cultured human gingival fibroblasts in vitro. Furthermore, periodontal pathogens, proinflammatory cytokines, N-acetylcysteine (NAC), U0126 and PD098059 were added to seek the possible regulatory mechanisms of PAI-1 expression. The results indicated that CsA increased PAI-1 expression on the fibroblasts (p<0.05). In CsA-treated fibroblasts, the additions of periodontal pathogens and proinflammatory cytokines significant enlarged the expression of PAI-1 (p<0.05), but the additions of NAC, U0126 and PD098059 reduced (p<0.05). Taken together, these results suggest that PAI-1 expression is significantly up-regulated in gingival tissues of cyclosporin A-treated patients. With periodontal pathogens and proinflammatory cytokines, PAI-1 expression was enhanced as compared with additions of CsA alone. The regulation of PAI-1 expression induced by cyclosporin A might be critically related with the intracellular glutathione and the ERK-MAPK pathway.

並列關鍵字

Cyclosporin A ； Plasminogen activator inhibitor-1 ； Gingival overgrowth ； Regulatory mechanism

參考文獻

Adams D, Davies G (1984). Gingival hyperplasia associated with cyclosporin A. A report of two cases. Br Dent J 157:89-90.

Alessi DR, Cuenda A, Cohen P, Dudley DT, Saltiel AR (1995). PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem 270(46):27489-94.

Allison MC, Pounder RE (1984). Cyclosporin for Crohn's disease. Lancet 1(8382):902-3.

Andreasen PA, Georg B, Lund LR, Riccio A, Stacey SN (1990). Plasminogen activator inhibitors: hormonally regulated serpins. Mol Cell Endocrinol 68(1):1-19.

Arora PD, Silvestri L, Ganss B, Sodek J, McCulloch CA (2001). Mechanism of cyclosporin-induced inhibition of intracellular collagen degradation. J Biol Chem 276(17):14100-9.

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第一型胞漿素原活化抑制劑與環孢靈誘發牙齦增生關係之探討

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