CAJ | 학술논문

通过研究高糖对牙周韧带细胞(priodontal ligament cells,PDLC)中转录因子Seleraxis表达的影响,探讨高糖抑制PDLC分化及糖尿病患者牙周病愈合延迟的分子机制.方法分别运用高糖型和低糖型DMEM培养基体外分离培养人PDLC,然后检测相应培养条件下碱性磷酸酶(ALP)活性和Scleraxis mRNA的表达.其中,ALP活性采用生物化学方法定量检测,Scleraxis mRNA的表达通过反转录聚合酶链反应(RT-PCR)技术半定量检测.结果与低糖型DMEM培养基培养的PDLC相比,高糖培养条件下PDLC的ALP活性下降,分别为0.218±0.012和0.113 ± 0.068;而Scleraxis的表达则增加,分别为0.611±0.205和0.973±0.055.经两独立样本t检验,ALP活性和Scleraxis表达的变化差异均有统计学意义(P<0.05).结论糖尿病患者的高血糖水平可能通过上调Scleraxis的表达来抑制PDLC向成骨细胞分化,进而影响牙周病的愈合.
통과연구고당대아주인대세포(priodontal ligament cells,PDLC)중전록인자Seleraxis표체적영향,탐토고당억제PDLC분화급당뇨병환자아주병유합연지적분자궤제.방법분별운용고당형화저당형DMEM배양기체외분리배양인PDLC,연후검측상응배양조건하감성린산매(ALP)활성화Scleraxis mRNA적표체.기중,ALP활성채용생물화학방법정량검측,Scleraxis mRNA적표체통과반전록취합매련반응(RT-PCR)기술반정량검측.결과여저당형DMEM배양기배양적PDLC상비,고당배양조건하PDLC적ALP활성하강,분별위0.218±0.012화0.113 ± 0.068;이Scleraxis적표체칙증가,분별위0.611±0.205화0.973±0.055.경량독립양본t검험,ALP활성화Scleraxis표체적변화차이균유통계학의의(P<0.05).결론당뇨병환자적고혈당수평가능통과상조Scleraxis적표체래억제PDLC향성골세포분화,진이영향아주병적유합.
Objective To approach the mechanisms of effects of high glucose on the differentiation of periodontal ligament cells (PDLC) by investigating the changes of Scleraxis mRNA expression in high glucose condition in vitro. Methods Human PDLC were cultured in high glucose medium (4500 mg/L glucese) and normal glucose medium (1000 mg/L glucose), respectively. High glucose was uesd to inhibit the ostengenic differentiation of PDLC. PDLC cultured in normal glucose medium served as controL. Alkaline phosphatase(ALP) activity, the early parameter of osteogenetic differentiation of cells and the expression of Scleraxis mRNA were detected in each group. ALP activity was measured colourmetrically by using nitrophenyl phosphate as a substrate and the expression of Scleraxis mRNA was analyzed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Results ALP activity of PDLC was lower in high glucose medium than in normal glucose medium, and the values were 0.113 ± 0.068 and 0.218 ± 0.012, respectively. However, the level of Scleraxis mRNA was quite higher in high glucose midium compared with in normal glucose medium, and the values were 0.973 ± 0.055 and 0.611 ± 0.205,respectively. The values of ALP activity and the expression of Scleraxis mRNA were significantly different between the two groups. Conclusions High glucose inhibited osteogenetic differentiation of PDLC and up-regulated Scleraxis expression. The adverse changes of Scleraxis expression and osteogenic differentiation of PDLC suggest that Scleraxis may regulate the esteogenic differentiation of PDLC negatively and the inhibition of high glucose on osteogenetic differentiation of PDLC may be regulated by Scleraxis in transcription level.