Nifedipine為一鈣離子阻斷劑，常廣泛用於治療高血壓及心血管疾病。然而，nifedipine有造成藥物性牙齦增生的副作用，容易去影響病人的口腔衛生、外觀等等。由於目前尚未找到能夠解決這個副作用的辦法，因此去了解nifedipine造成牙齦增生的分子機制就會顯得重要。目前大部分的研究都是著重在nifedipine作用在牙齦纖維母細胞時，其細胞因子或生長因子相關的表現，而較少研究透過幹細胞相關的基因來了解nifedipine造成牙齦增生上的分子機制。因此，我們利用human stem cell PCR array去預測標的基因，透過分子機制層面來了解藥物對牙齦纖維母細胞影響情形。我們首先用含有不同藥物濃度的培養基去培養牙齦纖維母細胞，分別培養一天、三天、五天、七天共四個時間點，從細胞存活率分析以及即時偵測細胞分析可知，在200 ng/ml的藥物濃度下，對細胞沒有毒殺性的影響，並在第七天細胞有增加的趨勢，利用CD44的mRNA表現量比較下，雖然與控制組比較沒有顯著的差異，但是在第七天相對其他天數有較高的表現，因此，我們最終選擇第七天作為時間點來做後續實驗。透過human stem cell PCR array結果，我們選出有相對明顯差異的基因作探討，包括ALDH2、COL2A1、FOXA2、WNT1，收集更多不同來源的細胞，透過即時定量聚合酶連鎖反應再次驗證mRNA表現，利用西方墨點法及免疫組織化學染色去觀察蛋白質分布，最後我們發現COL2A1在上述實驗中有相對明顯的表現，因此推論nifedipine造成牙齦增生的情形，COL2A1可能參與其中。

To solve the side-effect of nifedipine, it is important to realize the molecular mechanisms of nifedipine-induced gingival overgrowth. Mostly previous studies focus the cytokines or growth factors expressions on nifedipine treatment in fibroblasts, but there are few studies about stem cell markers in the mechanism of nifedipine-associated gingival overgrowth in gingival fibroblasts. Therefore, we cultured healthy gingival fibroblasts with different concentration of nifedipine medium and used human stem cell PCR array to predict target genes, western blots and immunohistochemistry to confirm the proteins expression. We found that 200 ng/mL concentration of DMSO and nifedipine did not influence on gingival fibroblasts proliferation. We also found that CD44 expressions of both groups were upregulated gradually as the time went. Therefore, we cultured the cells for 7 days to run human stem cell PCR array. PCR array results showed that most of genes were upregulated in gingival fibroblasts cultured in nifedipine-treated medium in heat map, but most gene changes were limited to the threshold of 2 folds. We collected more sources of gingival fibroblasts and chose the top of four genes to confirm the mRNA expressions and protein levels, including ALDH2、COL2A1、FOXA2 and WNT1. We found that COL2A1 had significant difference compared to control. We concluded that COL2A1 may participate in nifedipine-induced gingival overgrowth.

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