CAJ | 학술논문

背景：前期研究发现，周期性张应力在一定时间内可以诱导人牙周膜成纤维细胞增殖。目的：观察周期性张应力对人牙周膜成纤维细胞表达结缔组织生长因子的影响；明确JNK、p38MAPK、PI3K信号通路在周期性张应力诱导人牙周膜成纤维细胞表达结缔组织生长因子过程中的作用。方法：采用多通道细胞牵张应力加载系统对体外培养的人牙周膜成纤维细胞分别给予周期性张应力刺激1，6，12，24 h，并以未加力组为对照组。对加力12 h的细胞分别添加JNK、p38MAPK、PI3K信号通路特异性抑制剂预处理，并与未加抑制剂的细胞作对比。应用ELISA法检测培养细胞分泌到上清液中的结缔组织生长因子蛋白；应用荧光定量PCR技术检测细胞结缔组织生长因子mRNA的表达。结果与结论：加载周期性张应力组与对照组相比较，1 h人牙周膜成纤维细胞表达结缔组织生长因子开始增强、6 h表达明显增强，12 h达最高峰值、24 h表达开始下降。加入JNK信号通路特异性抑制剂后，人牙周膜成纤维细胞表达结缔组织生长因子出现下降；而加入p38MAPK信号通路和PI3K信号通路的特异性抑制剂后结缔组织生长因子表达未发生明显改变。提示在一定时间范围内，周期性张应力引起结缔组织生长因子 mRNA 与蛋白水平的表达与时间呈依赖性升高；其后随着时间的延长，结缔组织生长因子的表达则开始下降。周期性张应力通过 JNK 通路的介导调控人牙周膜成纤维细胞结缔组织生长因子的表达。
배경：전기연구발현，주기성장응력재일정시간내가이유도인아주막성섬유세포증식。목적：관찰주기성장응력대인아주막성섬유세포표체결체조직생장인자적영향；명학JNK、p38MAPK、PI3K신호통로재주기성장응력유도인아주막성섬유세포표체결체조직생장인자과정중적작용。방법：채용다통도세포견장응력가재계통대체외배양적인아주막성섬유세포분별급여주기성장응력자격1，6，12，24 h，병이미가력조위대조조。대가력12 h적세포분별첨가JNK、p38MAPK、PI3K신호통로특이성억제제예처리，병여미가억제제적세포작대비。응용ELISA법검측배양세포분비도상청액중적결체조직생장인자단백；응용형광정량PCR기술검측세포결체조직생장인자mRNA적표체。결과여결론：가재주기성장응력조여대조조상비교，1 h인아주막성섬유세포표체결체조직생장인자개시증강、6 h표체명현증강，12 h체최고봉치、24 h표체개시하강。가입JNK신호통로특이성억제제후，인아주막성섬유세포표체결체조직생장인자출현하강；이가입p38MAPK신호통로화PI3K신호통로적특이성억제제후결체조직생장인자표체미발생명현개변。제시재일정시간범위내，주기성장응력인기결체조직생장인자 mRNA 여단백수평적표체여시간정의뢰성승고；기후수착시간적연장，결체조직생장인자적표체칙개시하강。주기성장응력통과 JNK 통로적개도조공인아주막성섬유세포결체조직생장인자적표체。
BACKGROUND:Previous studies have found that cyclic tensile stress can induce the proliferation of human periodontal ligament fibroblasts in a certain time. OBJECTIVE: To observe the effect of cyclic tensile stress on the expression of connective tissue growth factor in human periodontal ligament fibroblasts, and to clarify the role of JNK, p38MAPK, PI3K pathway in the cyclic tensile stress-induced expression of connective tissue growth factor in human periodontal ligament fibroblasts. METHODS:Human periodontal ligament fibroblasts culturedin vitro were given stimulation of cyclic tensile stress in vitro for 1, 6, 12, 24 hours using multi-channel cel stretch stress loading system (experimental group). Control group was set up and not given any stimulation at the same time. The cels under stress application for 12 hours were given specific inhibitors of JNK, p38MAPK, PI3K respectively at the beginning, and were compared with those with no inhibitors. The concentrations of connective tissue growth factor in the culture supernatant were measured by ELISA. The mRNA expression of connective tissue growth factor in human periodontal ligament fibroblasts was detected by real-time RT-PCR. RESULTS AND CONCLUSION:Compared with the control group, the expression of connective tissue growthfactor in human periodontal ligament fibroblasts under cyclic tensile stress began to increase at 1 hour, increased manifestly at 6 hours, reached the peak at 12 hours, and then decreased at 24 hours. The expression of connective tissue growth factor was reduced by the specific inhibitors of JNK, while the specific inhibitors of p38 MAPK, PI3K had not the same effect. In a certain time, the cyclic tensile stress can induce a time-dependent increase in mRNA and protein levels of connective tissue growth factor. Along with the extending of time, the expression of connective tissue growth factor begins to decrease. Cyclic tensile stress can regulate expression of connective tissue growth factor in human periodontal ligament fibroblasts via the JNK pathway.