Abstract：
Periodontitis is a chronic inflammatory disease caused by Porphyromonas gingivalis and other bacteria, and human periodontal ligament stem cells (hPDLSCs) are a promising candidate for the treatment of periodontal supporting tissue defects. This study aimed to investigate the effect of 1α,25-dihydroxyvitamin D3 [1,25(OH)2VitD3] on osteogenic differentiation of hPDLSCs in an in vitro periodontitis model and whether it can improve inflammatory status. hPDLSCs were in vitro isolated and identified. After treatment with 1,25(OH)2VitD3 and ultrapure pure Porphyromonas gingivalis lipopolysaccharide (LPS-G), the viability of hPDLSCs was detected using Cell Counting Kit-8, the expressions of osteogenic markers and inflammatory genes using Western blotting and quantitative reverse transcription PCR (qRT-PCR), the levels of inflammatory factors in cells using enzyme linked immunosorbent assay (ELISA), and the fluorescence signal intensity of osteoblastic markers and inflammatory genes in cells using immunofluorescence assay. It was found that 1,25(OH)2VitD3 reversed the inhibition of hPDLSCs proliferation by LPS-G; LPS-G exhibited inhibitory effect on ALP, Runx2, and OPN expressions, and such inhibitory effect was significantly weakened when co-acting with 1,25(OH)2VitD3. Meanwhile, LPS-G upregulated the expressions of inflammatory genes IL-1β and Casp1, whereas 1,25(OH)2VitD3 antagonized such an effect and improved the inflammatory status. In conclusion, 1,25(OH)2VitD3 can reverse the inhibitory effect of LPS-G on hPDLSCs proliferation and osteogenic differentiation and suppress LPS-G-induced upregulation of inflammatory gene expressions.
摘要：
牙周炎是由牙龈卟啉单胞菌和其他细菌引起的慢性炎症性疾病，和人牙周膜干细胞（hPDLSCs）是治疗牙周支持组织缺损的有希望的候选者。本研究旨在探讨1α的作用，25-二羟维生素D3[1,25(OH)2VitD3]对体外牙周炎模型hPDLSCs成骨分化的影响及其是否能改善炎症状态.体外分离和鉴定hPDLSC。用1,25（OH）2VitD3和超纯牙龈卟啉单胞菌脂多糖（LPS-G）处理后，细胞计数试剂盒-8检测hPDLSCs的活力，蛋白质印迹和定量逆转录PCR（qRT-PCR）检测成骨标志物和炎症基因的表达，使用酶联免疫吸附试验（ELISA）检测细胞中炎症因子的水平，用免疫荧光法检测细胞中成骨细胞标志物和炎症基因的荧光信号强度。发现1,25(OH)2VitD3逆转了LPS-G对hPDLSCs增殖的抑制作用；LPS-G对ALP有抑制作用，Runx2和OPN表达式，当与1,25(OH)2VitD3共同作用时，这种抑制作用明显减弱。同时,LPS-G上调了炎症基因IL-1β和Casp1的表达，而1,25（OH）2VitD3则拮抗了这种作用并改善了炎症状态。总之,1,25(OH)2VitD3可以逆转LPS-G对hPDLSCs增殖和成骨分化的抑制作用，抑制LPS-G诱导的炎症基因表达上调。
