School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
Department of Nursing, Division of Basic Medical Sciences, Chronic Diseases and Health Promotion Research Center and Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Puzi City, Taiwan.
J Periodontal Res. 2021 Aug;56(4):804-818. doi: 10.1111/jre.12879. Epub 2021 Mar 17.
To investigate protective effects of Taiwanese green propolis (TGP) against high glucose-induced inflammatory responses in human gingival fibroblasts (HGFs) through NLRP3 inflammasome signaling pathway.
NLRP3 inflammasome has been implicated in the progression of both diabetes mellitus and periodontitis, suggesting a common potential therapeutic target for these diseases. Propolis is renowned for various biological activities, particularly anti-inflammation and antioxidant, representing a promising therapy for many conditions. However, underlying mechanisms remain unclear.
The cytotoxicity of TGP was evaluated by cell viability assay. The mRNA levels and protein expression or secretion of various inflammatory molecules and NLRP3 inflammasome-related molecules in high glucose-exposed HGFs with or without pretreatment of TGP (5 μg/ml) were determined by real-time PCR and western blot or specific kits, respectively. Intracellular and mitochondrial ROS measurements, NADPH oxidase activity determination, and subcellular fractions were performed to assess ROS generation. The transcriptional activity of NF-κB was measured by luciferase reporter kit. The signaling components were further differentiated using pharmacological inhibitors of ROS and small interfering RNAs of TLR2, TLR4, or NF-κB.
High glucose could induce IL-1β-driven inflammatory responses in HGFs via the activation of NLRP3 inflammasome regulated by TLR2/TLR4 coupled ROS in NF-κB-dependent manner. TGP had no adverse impact on the cell viability of HGFs at concentrations no greater than 10 μg/ml, and could exert inhibitory effects on high glucose-induced inflammatory responses via the interruption of NLRP3 inflammasome signaling pathway.
Taiwanese green propolis could elicit protective effects against IL-1β-driven inflammation in high glucose-exposed HGFs through TLR2/TLR4 combined ROS/NF-κB/NLRP3 inflammasome pathway.
通过 NLRP3 炎性小体信号通路，研究台湾绿蜂胶（TGP）对人牙龈成纤维细胞（HGF）高糖诱导的炎症反应的保护作用。
NLRP3 炎性小体参与糖尿病和牙周炎的进展，表明这些疾病有共同的潜在治疗靶点。蜂胶以其多种生物活性而闻名，特别是抗炎和抗氧化作用，是许多疾病的一种很有前途的治疗方法。然而，其潜在机制尚不清楚。
通过细胞活力测定评估 TGP 的细胞毒性。通过实时 PCR 和 Western blot 或特定试剂盒分别测定高糖暴露的 HGF 中各种炎症分子和 NLRP3 炎性小体相关分子的 mRNA 水平和蛋白表达或分泌，或用 TGP（5μg/ml）预处理后测定。通过测定细胞内和线粒体 ROS 水平、NADPH 氧化酶活性和亚细胞分数来评估 ROS 的产生。通过荧光素酶报告试剂盒测定 NF-κB 的转录活性。使用 ROS 和 TLR2、TLR4 或 NF-κB 的小干扰 RNA 的药理学抑制剂进一步区分信号转导成分。
高糖可通过 TLR2/TLR4 偶联 ROS 依赖 NF-κB 调节的 NLRP3 炎性小体诱导 HGF 中 IL-1β 驱动的炎症反应。TGP 在浓度不高于 10μg/ml 时对 HGF 的细胞活力没有不良影响，可通过阻断 NLRP3 炎性小体信号通路对高糖诱导的炎症反应发挥抑制作用。
台湾绿蜂胶可通过 TLR2/TLR4 联合 ROS/NF-κB/NLRP3 炎性小体通路，对高糖暴露的 HGF 中 IL-1β 驱动的炎症产生保护作用。
