Yi Min, Wang Guanglei, Niu Jianhua, Peng Minghui, Liu Yi
Department of Integrative Therapy, Shanghai Huangpu District 2nd Dental Disease Prevention and Treatment Institute, Shanghai 200001, P.R. China.
Department of Stomatology, Jiading District Central Hospital Affiliated to Shanghai University of Medicine and Health Sciences, Shanghai 201800, P.R. China.
Exp Ther Med. 2022 Apr;23(4):304. doi: 10.3892/etm.2022.11233. Epub 2022 Feb 22.
Periodontitis is a common chronic inflammatory oral disease. The objective of periodontal treatment is to control infection whilst regenerating damaged periodontal tissue. The present study aimed to determine the potential effects of pterostilbene (PTE), a representative stilbene compound, on the proliferation and differentiation of human periodontal ligament stem cells (hPDLSCs). Different concentrations (1, 5, 10 and 20 µM) of PTE were applied to hPDLSCs, after which Cell Counting Kit-8 and western blotting assays were performed to examine the protein levels of Ki67, PCNA, p-IκBα, IκBα, Bcl-2, Bax, cleaved caspase3. The effect of PTE on the release of inflammatory factors, including IL-1β, IL-6 and IL-10 was assessed by RT-qPCR. The apoptosis of TNF-α-induced hPDLSCs was evaluated by TUNEL assay and western blotting. Additionally, the role of PTE in hPDLSC mineralization was evaluated using alizarin red staining. The expression levels of mineralization indices, including RUNX2 and ALP were subsequently determined using western blotting. Subsequently, the target of PTE was predicted using TargetNet database and AutoDock v4.2 software and verified using western blotting. The results of the present study revealed that PTE promoted the proliferation of hPDLSCs in a concentration-dependent manner. Furthermore, PTE treatment decreased the release of inflammatory factors and alleviated the apoptosis of TNF-α-induced hPDLSCs. PTE was also demonstrated to promote the formation of mineral nodules in TNF-α-induced hPDLSCs. The Targetnet database, along with molecular docking, indicated that histone deacetylases (HDACs) were the probable targets of PTE upstream of regulating periodontitis. The results of western blotting implied that TNF-α significantly increased expression levels of HDAC2, 4, 6 and 8, whilst PTE treatment markedly decreased HDAC4, 6 and 8 expression in a concentration-dependent manner compared with the TNF-α group, which further confirmed these conclusions. In summary, results of the present study revealed that PTE promoted TNF-α-induced hPDLSC proliferation and differentiation, whilst alleviating inflammation and apoptosis. PTE also inhibited the expression of HDACs, which may be involved in the mechanism of periodontitis.
牙周炎是一种常见的慢性炎症性口腔疾病。牙周治疗的目标是控制感染，同时使受损的牙周组织再生。本研究旨在确定代表性芪类化合物紫檀芪（PTE）对人牙周膜干细胞（hPDLSCs）增殖和分化的潜在影响。将不同浓度（1、5、10和20 μM）的PTE应用于hPDLSCs，之后进行细胞计数试剂盒-8和蛋白质印迹分析，以检测Ki67、PCNA、p-IκBα、IκBα、Bcl-2、Bax、裂解的caspase3的蛋白质水平。通过RT-qPCR评估PTE对包括IL-1β、IL-6和IL-10在内的炎症因子释放的影响。通过TUNEL分析和蛋白质印迹评估TNF-α诱导的hPDLSCs的凋亡。此外，使用茜素红染色评估PTE在hPDLSC矿化中的作用。随后使用蛋白质印迹测定包括RUNX2和ALP在内的矿化指标的表达水平。随后，使用TargetNet数据库和AutoDock v4.2软件预测PTE的靶点，并通过蛋白质印迹进行验证。本研究结果显示，PTE以浓度依赖性方式促进hPDLSCs的增殖。此外，PTE处理降低了炎症因子的释放，并减轻了TNF-α诱导的hPDLSCs的凋亡。还证明PTE促进TNF-α诱导的hPDLSCs中矿结节的形成。Targetnet数据库以及分子对接表明，组蛋白脱乙酰酶（HDACs）是PTE在调节牙周炎上游的可能靶点。蛋白质印迹结果表明，TNF-α显著增加HDAC2、4、6和8的表达水平，而与TNF-α组相比，PTE处理以浓度依赖性方式显著降低HDAC4、6和8的表达，这进一步证实了这些结论。总之，本研究结果显示，PTE促进TNF-α诱导的hPDLSC增殖和分化，同时减轻炎症和凋亡。PTE还抑制HDACs的表达，这可能参与了牙周炎的机制。
