Yuan Ya-di, Miao Song, Xie Hao
School of Stomatology, Wuhan University, Wuhan 430079, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2008 Nov;43(11):668-70.
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.
Human PDLC were cultured in high glucose medium (4500 mg/L glucose) and normal glucose medium (1000 mg/L glucose), respectively. High glucose was used to inhibit the osteogenic 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 colorimetrically 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).
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 medium 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.
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 osteogenic differentiation of PDLC negatively and the inhibition of high glucose on osteogenetic differentiation of PDLC may be regulated by Scleraxis in transcription level.
通过体外研究高糖环境下牙周膜细胞（PDLC）中硬骨素（Scleraxis）mRNA表达的变化，探讨高糖对牙周膜细胞分化的影响机制。
将人牙周膜细胞分别培养于高糖培养基（葡萄糖浓度为4500 mg/L）和正常糖培养基（葡萄糖浓度为1000 mg/L）中。采用高糖抑制牙周膜细胞的成骨分化，以培养于正常糖培养基中的牙周膜细胞作为对照。检测每组细胞的碱性磷酸酶（ALP）活性（细胞成骨分化的早期指标）及Scleraxis mRNA的表达。采用磷酸对硝基苯酯作为底物，通过比色法测定ALP活性，采用半定量逆转录聚合酶链反应（RT-PCR）分析Scleraxis mRNA的表达。
高糖培养基中牙周膜细胞的ALP活性低于正常糖培养基，其值分别为0.113±0.068和0.218±0.012。然而，高糖培养基中Scleraxis mRNA水平明显高于正常糖培养基，其值分别为0.973±0.055和0.611±0.205。两组间ALP活性值及Scleraxis mRNA表达存在显著差异。
高糖抑制牙周膜细胞的成骨分化并上调Scleraxis表达。牙周膜细胞Scleraxis表达及成骨分化的不良变化表明，Scleraxis可能对牙周膜细胞的成骨分化起负性调节作用，高糖对牙周膜细胞成骨分化过程的抑制作用可能在转录水平受Scleraxis调控。
