Yang Ping, Shi Fenghua, Zhang Yanli
Department of Stomatology, AnTing Campus, The Third Affiliated Hospital of Naval Military Medical University, Shanghai 200438, P.R. China.
Department of Radiotherapy, AnTing Campus, The Third Affiliated Hospital of Naval Military Medical University, Shanghai 201805, P.R. China.
Exp Ther Med. 2022 Dec 22;25(2):74. doi: 10.3892/etm.2022.11773. eCollection 2023 Feb.
Periodontitis is the chronic inflammation of the periodontal tissue. The present study aimed to investigate the role of baricitinib, a Janus kinase (JAK)1/2 inhibitor, in periodontitis by using a lipopolysaccharide (LPS)-induced human periodontal ligament stem cell (PDLSC) model. The viability of PDLSCs stimulated by LPS was assessed in the presence of baricitinib by Cell Counting Kit-8 assay. The induction of oxidative stress was evaluated by detecting the intracellular reactive oxygen species (ROS) levels, superoxide dismutase (SOD) activity and glutathione (GSH) content. ELISA and reverse transcription-quantitative PCR were used to determine the levels of inflammatory factors TNF-α, IL-1β and IL-6. Alkaline phosphatase (ALP) activity and alizarin red staining were used to assess the osteogenic differentiation of PDLSCs. The expression levels of osteogenic differentiation- and JAK/signal transducer and activator of transcription (STAT) signaling-associated proteins were estimated with western blotting. RO8191, an agonist of the JAK/STAT pathway, was used to treat PDLSCs to investigate the regulatory mechanism of baricitinib. The results indicated that baricitinib elevated the LPS-induced decrease in cell viability. LPS-triggered oxidative stress and inflammation were inhibited by baricitinib, as demonstrated by the decreased levels of ROS, TNF-α, IL-1β, IL-6 and increased levels of SOD and GSH. In addition, baricitinib caused a marked elevation in ALP activity and mineralization ability of PDLSCs, as determined by the upregulated osteocalcin and Runt-related transcription factor 2 expression. Moreover, the expression levels of phosphorylated (p)-JAK1, p-JAK2 and p-STAT3 were downregulated by baricitinib in a dose-dependent manner. Furthermore, addition of RO8191 restored the effect of baricitinib on the induction of oxidative stress, inflammation and osteogenic differentiation of PDLSCs exposed to LPS. Collectively, these findings suggested that baricitinib alleviated oxidative stress and inflammation and promoted osteogenic differentiation of LPS-induced PDLSCs by inhibiting JAK/STAT signaling.
牙周炎是牙周组织的慢性炎症。本研究旨在通过使用脂多糖(LPS)诱导的人牙周膜干细胞(PDLSC)模型,研究Janus激酶(JAK)1/2抑制剂巴瑞替尼在牙周炎中的作用。通过细胞计数试剂盒-8法在有巴瑞替尼存在的情况下评估LPS刺激的PDLSC的活力。通过检测细胞内活性氧(ROS)水平、超氧化物歧化酶(SOD)活性和谷胱甘肽(GSH)含量来评估氧化应激的诱导情况。采用酶联免疫吸附测定(ELISA)和逆转录定量聚合酶链反应(RT-qPCR)来测定炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的水平。使用碱性磷酸酶(ALP)活性和茜素红染色来评估PDLSC的成骨分化。用蛋白质印迹法估计成骨分化和JAK/信号转导及转录激活因子(STAT)信号相关蛋白的表达水平。使用JAK/STAT途径的激动剂RO8191处理PDLSC,以研究巴瑞替尼的调节机制。结果表明,巴瑞替尼提高了LPS诱导的细胞活力下降。巴瑞替尼抑制了LPS引发的氧化应激和炎症,表现为ROS、TNF-α、IL-1β、IL-6水平降低,SOD和GSH水平升高。此外,通过上调骨钙素和 runt相关转录因子2的表达确定,巴瑞替尼使PDLSC的ALP活性和矿化能力显著提高。此外,巴瑞替尼以剂量依赖性方式下调磷酸化(p)-JAK1、p-JAK2和p-STAT3 的表达水平。此外,添加RO8191恢复了巴瑞替尼对暴露于LPS的PDLSC的氧化应激诱导、炎症和成骨分化的影响。总的来说,这些发现表明,巴瑞替尼通过抑制JAK/STAT信号减轻氧化应激和炎症,并促进LPS诱导的PDLSC的成骨分化。