細胞核因子κB活化因子受體配體與噬骨細胞抑制因子在細菌性脂多醣誘發根尖病變的老鼠模型中的長期變化

Long-term sequential receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) expression in lipopolysaccharide-induced rat periapical lesions

莊富雄(Fu-Hsiung Chuang)

指導教授 ： 林英助

共同指導教授 ： 陳玉昆(Yuk-Kwan Chen)

高雄醫學大學/口腔醫學院/牙醫學研究所/博士(2013年)

https://doi.org/10.6832/KMU.2013.00008

摘要

RANKL/RANK/OPG系統，包括細胞核因子κB活化因子受體配體（Receptor Activator of NF Kappa B Ligand，RANKL）、細胞核因子κB活化因子受體（Receptor activator of NF Kappa B，RANK)與噬骨細胞抑制因子（osteoprotegerin，OPG)，是骨頭代謝疾病非常重要的角色，不過，在根尖周圍疾病的相關資訊仍極為缺乏。本研究的目的，即是要分析RANKL-OPG在整個根尖周圍炎的形成與發展過程中，長期連續的特異性影響。實驗以72隻Wistar大鼠，分成八組實驗組與一組對照組，使用細菌性脂多醣（LPS）誘發形成根尖病變，每週犧牲一組，共八週，利用放射線檢查測量其病灶面積；以即時定量聚合酶鏈鎖反應測量根尖組織RANKL-OPG的平均mRNA表現量；使用免疫組織化學法測定RANKL-OPG的蛋白質表現；噬骨細胞活性則以酵素免疫化學法決定，最後以酵素連結免疫吸附分析法來分析血清中RANKL與OPG的濃度變化。結果顯示，根尖病灶平均相對基因表現量中，mrRANKLmRNA與mrRANKLmRNA/mrOPGmRNA比例的高低，與病灶破壞面積大小均呈現顯著正相關，mrOPGmRNA高低則與病灶破壞面積大小呈現顯著負相關，而mrRANKLmRNA/mrOPGmRNA比例則於第七、八週達到顯著高峰（p<0.05）。此變化也和組織學、酵素組織化學與免疫組織化學的檢查結果一致。另外，血中的RANKL-OPG濃度也與比例與根尖RANK-OPG表現量高低呈顯著正相關。由以上結果證實，根尖組織RANKL-OPG的變化的確與根尖病灶的形成與擴張相關，也影響老鼠全身性RANKL-OPG濃度的變化。未來則期望應用實驗模式的潛能，藉由監測RANKL-OPG的變化來於評估各種實驗性的治療結果。

關鍵字

細胞核因子κB活化因子受體配體 ； 噬骨細胞抑制因子 ； 細菌性脂多醣 ； 根尖周圍炎

並列摘要

The RANKL/RANK/OPG system, including receptor activator of NF-κB ligand (RANKL) and its receptors, receptor activator of NF-κB (RANK) and osteoprotegrin (OPG), play a crucial role in bone metabolic diseases. However, relatively little information is available in the periapical diseases. The aim of this study was to analyze the specific involvement of the long term sequential expression of RANKL-OPG system in the progress and development of apical periodontitis. 72 Wistar rats were divided into eight experimental groups and one control group. After the induction of apical lesions by lipopolysaccharide, the mean area of periapical lesion (mAREAperiapical) was measured by radiographic imaging. Relative mean mRNA expression of RANKL (mrRANKLmRNA) and OPG (mrOPGmRNA) were measured in all tissue sections every week from week 1 to week 8 by real-time quantitative reverse transcription polymerase chain reaction. RANKL and OPG protein will be determined by immunohistochemistry. Osteoclasts will be identified by enzyme histochemistry. The concentrations of RANKL-OPG in the serum were analyzed by enzyme-linked immunosorbent assay (ELISA). The results showed there was a positive significantly correlation between the mAREAperiapical and mrRANKLmRNA as well as mrRANKLmRNA /mrOPGmRNA ratio, and the mAREAperiapical was negatively correlated with mrOPGmRNA. Expression ratio of RANKL against OPG peaked at 3, 7 and 8 weeks. The sequential changes in the mRNA expression of RANKL and OPG were largely compatible with the occurrence of osteoclasts histologically and enzymes histochemically as well as immunohistochemistry. The concentrations of RANKL and OPG in serum also changed during the local infection. It showed a positive significantly correlation with the level of mrRANKLmRNA and mrOPGmRNA in the apical tissue. In conclusion, we suggested that the RANKL-OPG system do be responsible for the initiation and expansion of periapical diseases. The change of RANKL-OPG concentration systemically was also correlated the level of expression of RAMKL-OPG in periapical periodontitis. The long-term findings further extend the potential application of the model for future experimental trials to evaluate the treatment outcome by monitoring the change in RANKL expression or the RANKL/OPG ratio in the samples.

並列關鍵字

receptor activator of NF-κB ligand (RANKL) ； osteoprotegerin (OPG) ； Lipopolysaccharide (LPS) ； apical periodontitis

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