Qin Han, Xu Hong-Zhi, Gong Yong-Qing
a Department of Stomatology , Lianyungang Affiliated Hospital of Xuzhou Medical University , Liangyungang , Jiangsu Province , China.
Mol Membr Biol. 2016 Sep-Dec;33(6-8):138-144. doi: 10.1080/09687688.2017.1400601. Epub 2017 Nov 23.
The objective of the present work was to investigate a possible mechanism of NF-κB signaling pathway and autophagy in the regulation of osteoblast differentiation, and provide experimental basis for the study of tooth eruption disorder.
Mouse osteoblast-like (MC3T3-E1) cells were inoculated with a cell density of 70%. According to the grouping experimental design, Western blot and monodansylcadaverine (MDC) detection were conducted after dosing for 24 h. The cells were divided into the following five groups: blank control group; 6.25 µg/mL SN50 group; 12.5 µg/mL SN50 group; 25 µg/mL SN50 group and 50 µg/mL SN50 group.
Western blot analysis revealed that the expression of LC3 protein was present in the blank control group; 6.25 µg/mL SN50 group; 12.5 µg/mL SN50 group and 50 µg/mL SN50 group, with no significant differences among these groups. However, the expression of LC3 protein was significantly lower in the 25 µg/mL SN50 group. MDC detection showed that, in the blank control group; 6.25 µg/mL SN50 group; 12.5 µg/mL SN50 group and 50 µg/mL SN50 group, there was obvious green fluorescence in the cytoplasm of the osteoblasts. However, in the 25 µg/mL SN50 group, it was found that there were significantly fewer green fluorescent particles.
The osteoblast itself had a strong function of autophagy. The appropriate concentration of SN50 in blocking the NF-κB pathway of the osteoblast was associated with the obvious inhibition of autophagy. However, the relationship between NF-κB signaling pathway and autophagy in the process of tooth eruption requires further study.
本研究旨在探讨核因子κB（NF-κB）信号通路和自噬在成骨细胞分化调控中的可能机制，为牙萌出紊乱的研究提供实验依据。
以70%的细胞密度接种小鼠成骨样（MC3T3-E1）细胞。根据分组实验设计，给药24小时后进行蛋白质免疫印迹法（Western blot）和单丹磺酰尸胺（MDC）检测。细胞分为以下五组：空白对照组；6.25μg/mL SN50组；12.5μg/mL SN50组；25μg/mL SN50组和50μg/mL SN50组。
蛋白质免疫印迹法分析显示，空白对照组、6.25μg/mL SN50组、12.5μg/mL SN50组和50μg/mL SN50组均有微管相关蛋白1轻链3（LC3）蛋白表达，各组间差异无统计学意义。然而，25μg/mL SN50组LC3蛋白表达明显降低。MDC检测显示，空白对照组、6.25μg/mL SN50组、12.5μg/mL SN50组和50μg/mL SN50组的成骨细胞胞质中有明显的绿色荧光。然而，在25μg/mL SN50组中，发现绿色荧光颗粒明显减少。
成骨细胞自身具有较强的自噬功能。适当浓度的SN50阻断成骨细胞的NF-κB通路与自噬明显受抑制有关。然而，NF-κB信号通路与牙萌出过程中自噬的关系尚需进一步研究。
