熱休克蛋白47與環孢靈誘發牙齦增生之探討

環孢靈(cyclosporin A)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的副作用，而造成牙齦增生的主要原因是牙齦細胞外基質產生異常堆積的結果。熱休克蛋白47主要是存在於產生膠原細胞的內質網內，負責產生type I的膠原蛋白及膠原蛋白的活化，並與一些纖維化的疾病有著密切的關係，然而目前有關與環孢靈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，探討正常牙齦與環孢靈誘發增生牙齦中熱休克蛋白47的表達；結果發現熱休克蛋白47表達在牙齦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，可以觀察到較多的熱休克蛋白47表達。另培養人類正常牙齦纖維母細胞，利用逆反轉錄聚合酶連鎖反應、西方點墨法，探討加入環孢靈後，熱休克蛋白47是否會被誘發表現；另外，本研究再多加入牙周致病菌 Aggregatibacter actinomycetemcomitans、第一型白介質素和MEK Pathway的抑制素U0126，探討環孢靈影響熱休克蛋白47表現的可能機轉。結果顯示，人類正常牙齦纖維母細胞加入環孢靈後，不論在mRNA或蛋白質的表現層次上，熱休克蛋白47的表現都是增加的(p<0.05)；另一方面，與單純加入環孢靈的結果相較，再多加入牙周致病菌或發炎激素，對於熱休克蛋白47的表現產生了上升的趨勢(p<0.05)；加入U0126則熱休克蛋白47明顯被抑制(p<0.05)。綜合研究結果顯示，環孢靈誘發熱休克蛋白47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，熱休克蛋白47的表現會更明顯，此外，環孢靈影響熱休克蛋白47表現的調控機轉，可能與細胞內MEK pathway有關。

關鍵字

環孢靈；牙齦增生；熱休克蛋白 47 ；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). Heat shock protein (HSP) 47, a collagen-specific molecular chaperone, is involved in the processing and/or secretion of procollagen. HSP47 is consistently and dramatically upregulated in a variety of fibrotic diseases. The aim of this study was to compare HSP47 expression in normal gingival tissues and CsA induced gingival overgrowth specimens and further explore the potential mechanism that may lead to induce HSP47 expression. However, the correlation between HSP47 protein and CsA-induced gingival overgrowth was little known, and it was the main purpose we investigated in this study. Immunohistochemistry was used to compare HSP47 expression between gingival tissues obtained from normal patients and CsA-treated patients in vivo. The evaluations displayed that greater expression of HSP47 was observed on fibroblasts and inflammatory cells in CsA-induced hyperplasia gingive. Reverse transcriptase-polymerase chain reaction、Western blotting were used to determine the effects of CsA on the expression of HSP47 in cultured human gingival fibroblasts in vitro. Furthermore, proinflammatory cytokines interleukin-1α，periodontal pathogen Aggregatibacter actinomycetemcomitans and MEK inhibitor U0126 were added to seek the possible regulatory mechanisms of HSP47 expression. The results indicated that CsA increased expression on the fibroblasts (p<0.05). In CsA-treated fibroblasts, the additions of periodontal pathogens and proinflammatory cytokines significant enlarged the expression of HSP47 (p<0.05), but the additions of U0126 reduced (p<0.05). Taken together, these results suggest that HSP47 expression is significantly up-regulated in gingival tissues of cyclosporin A-treated patients. With periodontal pathogens and proinflammatory cytokines, HSP47 expression was enhanced as compared with additions of CsA alone. The regulation of HSP47 expression induced by cyclosporin A might be critically related with the MEK pathway.

並列關鍵字

Cyclosporin A ； Heat shock protein 47 ； 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.

Chae HJ, Ha MS, Yun DH, Pae HO, Chung HT, Chae SW, Jung YK, Kim HR. Mechanism of cyclosporine-induced overgrowth in gingival. J Dent Res 2006; 85: 515-519.

Chang YC, Yang SF, Lai CC, Liu JY, Hsieh YS. Regulation of matrix metalloproteinases production by cytokines, pharmacological agents, and periodontal pathogens in human periodontal ligament fibroblast cultures. J Periodont Res 2002; 37: 196-203.

Chang YC, Tsai CH, Yang SH, Liu CM, Chou MY. Induction of cyclooxygenase-2 mRNA and protein expression in human gingival fibroblasts stimulated with nicotine. J Periodont Res 2003; 38: 496-501.

Chang YC, Lai CC, Lin LF, Ni WF, Tsai CH. The upregulation of heme oxygenase-1 expression in human gingival fibroblasts stimulated with nicotine. J Periodont Res 2005; 40: 252-257.

國際替代計量

熱休克蛋白47與環孢靈誘發牙齦增生之探討

全文下載

主題瀏覽