Bai Si-Yu, Chen Yue, Dai Hong-Wei, Huang Lan
Dept. of Orthodontics, Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2019 Apr 1;37(2):162-167. doi: 10.7518/hxkq.2019.02.007.
The purpose of this study is to investigate the potential effects of sclerostin (SOST) on the biological funtions and related mechanisms of cementoblasts under mechanical stress.
OCCM-30 cells were treated with varying doses of SOST (0, 25, 50, and 100 ng·mL⁻¹) and were loaded with uniaxial compressive stress (2 000 μ strain with a frequency of 0.5 Hz) for six hours. Western blot was utilized to detect the expressions of β-catenin, p-smad1/5/8, and smad1/5/8 proteins. Alkaline phosphatase (ALP) activity was determined, and reverse transcription polymerase chain reaction was used to measure the expressions of runt-related transcription factor 2 (Runx-2), osteocalcin (OCN), bone sialoproteins (BSP), receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) mRNA.
RESULTS: The expression of p-smad 1/5/8 was significantly downregulated with increasing SOST. β-catenin and smad1/5/8 exhibited no difference. ALP activity decreased under mechanical compressive stress with increasing SOST concentrations. Runx-2 expression was reduced with increasing SOST concentrations, and a similar trend was observed for the BSP and OCN expressions. When the SOST concentration was enhanced, RANKL expression gradually increased, whereas the expression of OPG decreased.
Under mechanical comprehensive stress, SOST can adjust the bone morphogenetic protein (BMP) /smad signal pathway. Osteosclerosis inhibits the mineralization of cementoblasts under mechanical compressive stress, which may be achieved by inhibiting the expressions of osteogenesis factors (Runx2, OCN, BSP, and others) and by promoting the ratio of cementoclast-related factors (RANKL/OPG) through BMP signal pathways.
本研究旨在探讨硬化蛋白(SOST)对机械应力作用下成牙骨质细胞生物学功能及相关机制的潜在影响。
用不同剂量的SOST(0、25、50和100 ng·mL⁻¹)处理成牙骨质细胞系(OCCM-30),并施加单轴压缩应力(2 000 μ应变,频率为0.5 Hz)6小时。采用蛋白质免疫印迹法检测β-连环蛋白、磷酸化Smad1/5/8和Smad1/5/8蛋白的表达。测定碱性磷酸酶(ALP)活性,并用逆转录聚合酶链反应检测 runt相关转录因子2(Runx-2)、骨钙素(OCN)、骨唾液蛋白(BSP)、核因子κB受体活化因子配体(RANKL)和骨保护素(OPG)mRNA的表达。
随着SOST剂量增加,磷酸化Smad1/5/8的表达显著下调。β-连环蛋白和Smad1/5/8的表达无差异。在机械压缩应力下,随着SOST浓度增加,ALP活性降低。随着SOST浓度增加,Runx-2表达降低,BSP和OCN表达也呈现类似趋势。当SOST浓度升高时,RANKL表达逐渐增加,而OPG表达降低。
在机械复合应力作用下,SOST可调节骨形态发生蛋白(BMP)/Smad信号通路。骨硬化蛋白在机械压缩应力下抑制成牙骨质细胞矿化,这可能是通过抑制成骨因子(Runx2、OCN、BSP等)的表达,并通过BMP信号通路促进破牙骨质细胞相关因子(RANKL/OPG)的比例来实现的。