CAJ | 학술논문

目的：??研究过量氟对体外培养大鼠成釉细胞内钙超载及细胞凋亡的影响。方法??取大鼠成釉细胞系HAT-7细胞，分别加入不同浓度（0、0.4、0.8、1.6、3.2、6.4?mmol·L-1）的氟化钠培养液，培养48?h后，采用Cell?Counting?Kit?8（CCK-8）试剂盒检测各组细胞的活性，流式细胞术分析氟对细胞凋亡的影响，激光扫描共聚焦显微镜、Western?blot试验和实时荧光定量聚合酶链反应技术检测过量氟诱导大鼠成釉细胞内Ca2+浓度和钙网蛋白表达的变化。结果?氟化钠浓度高于1.6?mmol·L-1时，可抑制成釉细胞的活性，成釉细胞内Ca2+浓度升高，钙网蛋白表达上调，细胞早期凋亡数量增加，并且随着浓度的增加，细胞凋亡的数量也随之增加。结论??过量氟可引起成釉细胞内钙超载，诱导成釉细胞凋亡。
목적：??연구과량불대체외배양대서성유세포내개초재급세포조망적영향。방법??취대서성유세포계HAT-7세포，분별가입불동농도（0、0.4、0.8、1.6、3.2、6.4?mmol·L-1）적불화납배양액，배양48?h후，채용Cell?Counting?Kit?8（CCK-8）시제합검측각조세포적활성，류식세포술분석불대세포조망적영향，격광소묘공취초현미경、Western?blot시험화실시형광정량취합매련반응기술검측과량불유도대서성유세포내Ca2+농도화개망단백표체적변화。결과?불화납농도고우1.6?mmol·L-1시，가억제성유세포적활성，성유세포내Ca2+농도승고，개망단백표체상조，세포조기조망수량증가，병차수착농도적증가，세포조망적수량야수지증가。결론??과량불가인기성유세포내개초재，유도성유세포조망。
Objective To study the effect of excessive fluoride on calcium overload and apoptosis in cultured rat ameloblasts in vitro. Methods Logarithmic-phase ameloblasts (HAT-7) were treated with 0, 0.4, 0.8, 1.6, 3.2, and 6.4 mmol·L-1 sodium fluoride (NaF) solution. Cell activities were detected by using a Cell Counting Kit 8 (CCK-8) assay after 48 h of treatment. The effect of fluoride on cell apoptosis was analyzed by using flow cytometry. Excessive fluoride-induced calcium concentration and calreticulin expression changes in ameloblasts were detected by using laser scanning confocal microscopy, Western blot analysis, and real-time quantitative polymerase chain reaction. Results NaF inhibited ameloblast activity at 1.6, 3.2, and 6.4 mmol·L?1 (dose-dependent) after 48 h of induction. The Ca2+ fluorescence intensity of HAT-7 cells incubated with 1.6 and 3.2 mmol·L?1 NaF was higher than that in the control group. The fluoride-induced early-stage apoptosis of ameloblasts after 48 h of induction and the early-stage apoptosis rate was positively correlated with fluoride concentration. Calreticulin mRNA expression in HAT-7 cells was higher than that in the control group after 48 h of incubation with 0.8, 1.2, and 1.6 mmol·L?1 NaF. Conclusion Excessive fluoride-induced calcium overload in ameloblasts and further caused endoplasmic reticulum stress-mediated apoptosis.