- 學位論文
以免疫組織化學染色法觀察牙齦組織中活化態 caspase-3 之表現分布狀況因牙周炎而不同
Active Caspase‐3 Expressed Differently in Periodontal Inflamed Gingival Using Immunohistochemical Analysis Method
摘要
背景 牙周病雖然為一種感染性疾病，但近年來研究都顯示，調 控宿主的發炎反應可能是避免宿主免疫反應加重牙周組織傷害的關 鍵。Casepase-3 是與凋亡相關的半胱胺酸脢家族成員，許多研究顯示 active caspase-3 的出現是凋亡的重要指標。細胞藉由凋亡機制 死亡，可免於細胞壞死造成的細胞激素外溢和毒素累積的傷害。證據 顯示，牙周炎病灶中凋亡與發炎反應之間的不平衡可能是牙周病破壞 的機轉之一。 研究目的 本研究的目的為藉由免疫組織化學染色法分析牙周 發炎組織與牙周健康組織之中，active caspase-3 的表現是否不同， 以觀察牙周發炎時凋亡現象是否有所改變。 材料與方法 由慢性牙周病病患發炎處取得之 18 例發炎牙齦組 織為實驗組，患者對照組為其中 8 位也有取得相對健康區域組織之病 患，另有無牙周病之健康對照組織 7 例。組織經石蠟包埋，連續切片 後，進行免疫組織化學染色法之 LSAB 法染色，並在顯微鏡下觀察 active caspae-3 的染色表現。 結果 active caspase-3 的染色範圍在健康對照組、患者對照 組和患者發炎組之間，兩兩具顯著差異(P=0.0001)。在染色強度部 份，三組之間也是認兩組之間有顯著的染色差異(P<0.0001)。整個 active caspase-3 的分布情形為患者對照組顯著高於其他兩組。 結論 active caspase-3 在牙周病患者發炎組織之中表現有比 健康患者低的趨勢，而在患者相對較健康的區域，雖然臨床發炎現象 較低，但是 active casepase-3 卻高度表現，顯示在牙周病灶高度發 炎時，凋亡因子 active caspase-3 的表現可能受到抑制調控，當發 炎下降時，細胞凋亡因子 active caspase-3 就開始高度表現，可見 牙周炎組織之中發炎與凋亡之間確實有關，需要進一步實驗證實其因 果關係。
並列摘要
Background Although periodontitis is a kind of infectious disease, studies indicate that regulation of host own inflammatory reaction may be a key pathway to prevent periodontal damage that caused by host immune reaction. Caspase-3 is a member of caspase family and is involved in cell apoptosis. Many studies suggest that the expression of active caspase-3 may be one of the apoptosis key markers. Cell death through apoptosis pathway can prevent the damage cause by cell necrosis, including release of cytokines and cytotoxins. The imbalance between apoptosis and inflammation may be a mechanism of periodontal pathogenesis. Objective The aim of this study was to investigate active caspase-3 expression in periodontal tissue from healthy sites and inflammatory sites, to find out if there is different apoptotic activity in periodontal healthy and inflammatory sites. Material and Methods Eighteen inflammatory tissues were obtained from chronic periodontitis patients, 8 relatively healthy tissues were also obtained from these patients to act as control sites of periodontal diseased patients, and 7 gingival tissues from non periodontitis individuals were included in this study. Tissue samples were paraffin fixed and cut into sections, and examined microscopically after immunohistochemical staining -LSAB method. Result The stained area of active caspase-3 was different(P=0.0001) among groups of those three groups of healthy contol, patient control, and inflammatory group . The stained intensity also was statistically significant different between patient control group and the other two groups(P<0.0001). The expression stained power of active caspase-3 in patient control group is significantly higher than the other two groups(P<0.0001). Conclusion The statistically less expression of active caspase-3 in tissues of inflammatory group and the abundant expression of active caspase-3 in tissues of patient control group suggest that apoptostic activity may be reduced in highly inflamed periodontal tissue but highly expressed in relatively healthy periodontal tissue of periodontitis patients. This result highly suggest that there is some relationship between inflammation and apoptosis in periodontal tissue. However, further studies with a larger sample size are needed to clarify this relationship in periodontal disease.