Department of Orthodontics, Stomatological Hospital, General Hospital of Ningxia Medical University, Yinchuan 750004, PR China.
Department of Prosthodontics, Yinchuan Stomatological Hospital, Yinchuan 750004, PR China.
Biomed Pharmacother. 2023 Oct;166:115391. doi: 10.1016/j.biopha.2023.115391. Epub 2023 Sep 6.
Orthodontic tooth movement (OTM) is a typical treatment that corrects malaligned teeth by applying mechanical forces. However, mechanical overload often leads to damage of PDL fibroblasts. Sanhuang decoction (SHD) is commonly used to inhibit inflammation and oxidative stress. However, the mechanism of SHD for OTM treatment is still unclear. Therefore, this study attempts to explore the underlying mechanism through relevant experiments.
In the present paper, we established a OTM rat model and further explored the effects of SHD on the PDL of OTM rats. The OTM model and effects of SHD were determined by micro-CT, and the PDL pathological changes, PDL width and capillaries in PDL were observed by H&E staining. Subsequently, the ROS levels in PDL was determined using flow cytometry analysis with DCFH-DA staining, MDA contents and antioxidative enzymes activities were also measured using commercial kits. Furthermore, the autophagy of PDL fibroblasts and proteins in the PI3K/Akt/mTOR pathway were detected using immunoluminescence, qPCR and western blotting assays.
The results showed SHD treatment can alleviate the decrease of PDL cells and capillaries induced by OTM, and improve the MDA and ROS levels in PDL, as well as enhance the activities of SOD and GSH-Px. Further experiments indicated SHD decreased the autophagy levels of PDL fibroblasts via promoting the phosphorylation levels of mTOR, PI3K and Akt proteins.
SHD inhibited autophagy of periodontal ligament fibroblasts during orthodontic tooth movement by inhibiting oxidative stress via activating PI3K-Akt-mTOR pathway. Our present findings suggested SHD treatment would be useful for management of the possible disorders occurs in orthodontic tooth movement therapy.
正畸牙齿移动（OTM）是通过施加机械力来矫正错位牙齿的典型治疗方法。然而，机械过载常导致牙周膜成纤维细胞损伤。三黄汤（SHD）常用于抑制炎症和氧化应激。然而，SHD 治疗 OTM 的机制尚不清楚。因此，本研究试图通过相关实验探讨其潜在机制。
本文建立了 OTM 大鼠模型，并进一步探讨了 SHD 对 OTM 大鼠牙周膜的影响。通过 micro-CT 确定 OTM 模型和 SHD 的作用，通过 H&E 染色观察牙周膜的病理变化、牙周膜宽度和牙周膜中的毛细血管。随后，用 DCFH-DA 染色通过流式细胞术分析测定牙周膜中的 ROS 水平，用商业试剂盒测定 MDA 含量和抗氧化酶活性。此外，用免疫荧光、qPCR 和 Western blot 检测牙周膜成纤维细胞的自噬和 PI3K/Akt/mTOR 通路中的蛋白。
结果表明，SHD 治疗可减轻 OTM 引起的牙周膜细胞和毛细血管减少，并改善牙周膜中的 MDA 和 ROS 水平，同时增强 SOD 和 GSH-Px 的活性。进一步的实验表明，SHD 通过促进 mTOR、PI3K 和 Akt 蛋白的磷酸化水平来降低牙周膜成纤维细胞的自噬水平。
SHD 通过激活 PI3K-Akt-mTOR 通路抑制氧化应激，抑制正畸牙齿移动过程中的牙周膜成纤维细胞自噬。我们的研究结果表明，SHD 治疗可能有助于管理正畸牙齿移动治疗中可能出现的疾病。
