- 學位論文
探討短鏈脂肪酸丁酸鈉誘發造牙骨質母細胞株細胞毒性的研究
The study of exploring the short chain fatty acid sodium butyrate-induced cytotoxicity on cementoblast
摘要
造成牙周感染的革蘭氏陰性厭氧菌會產生一種名為丁酸（butyric acid）的短鏈脂肪酸（short chain fatty acid，簡稱SCFA），其會造成造骨母細胞等牙周相關細胞的毒性作用，由文獻回顧鮮少有研究探討丁酸鈉(sodium butyrate)對造牙骨質母細胞的影響。因此本研究培養老鼠造牙骨質母細胞(OCCM.30)，探討丁酸鈉對於OCCM.30細胞的毒性作用。試驗以不同濃度的丁酸鈉( 0、2、4、8、16 mM)，作用於OCCM.30細胞，利用MTT試驗觀察細胞的存活率，並以Annexin V-FITC/PI雙染色實驗，探討丁酸鈉對OCCM.30細胞週期的影響；另進一步研究探討丁酸鈉對OCCM.30與細胞凋亡蛋白胱天蛋白酶(caspase)訊號傳遞路徑的相關性。實驗結果顯示，丁酸鈉在濃度4 mM即會明顯地抑制造牙骨質母細胞的存活率，隨劑量的增加而增加，具有統計學上的意義(p＜0.05)，丁酸鈉在濃度4 mM即會造成OCCM.30細胞sub-G1週期停滯與細胞凋亡，另以西方墨點法發現會誘發pro-caspase-8，pro-caspase-9, pro-caspase-3的活性(p＜0.05)，並且會誘導MAPK路徑中磷酸化ERK、JNK和p38蛋白的表現。根據初步的實驗結果，發現丁酸鈉對造牙骨質母細胞具有細胞毒性，並與細胞凋亡的路徑胱天蛋白酶家族有關連性。
並列摘要
Gram-negative anaerobic bacteria causing periodontal infections produce a product called butyric acid, a short-chain fatty acid that exerts toxic effects on periodontal-related cells, including osteoblasts. However, there is limited research on the effects of sodium butyrate on cementoblast. Therefore, this study aimed to investigate the toxicity of sodium butyrate on mouse cementoblast (OCCM.30). Different concentrations of sodium butyrate (0, 2, 4, 8, 16 mM) were used to treat OCCM.30 cells. Cell viability was assessed using the MTT assay, and the effects of sodium butyrate on the cell cycle of OCCM.30 cells were examined using Annexin V-FITC/PI double staining. Furthermore, the relationship between sodium butyrate and the apoptotic protein caspase signaling pathway in OCCM.30 cells was investigated. The experimental results showed that sodium butyrate significantly inhibited the viability of odontoblastic cells at a concentration of 4 mM, with a dose-dependent effect (p < 0.05). Sodium butyrate induced sub-G1 cell cycle arrest and apoptosis in OCCM.30 cells at a concentration of 4 mM, as evidenced by Western blot analysis showing increased activity of pro-caspase-8, pro-caspase-9, and pro-caspase-3 (p < 0.05),and induced the expression of phosphorylated ERK, JNK and p38 proteins in the MAPK pathway. Based on the preliminary findings, sodium butyrate was found to exhibit cytotoxicity on odontoblastic cells and was associated with the caspase family pathway involved in cell apoptosis.