Department of Oral Anatomy and Physiology, School and Hospital of Stomatology, China Medical University, Liaoning Provincial Key Laboratory of Oral Disease, Shenyang, 110002, China.
Department of Science Experiment Center of China Medical University, Shenyang, 110122, China.
Arch Oral Biol. 2021 May;125:105110. doi: 10.1016/j.archoralbio.2021.105110. Epub 2021 Mar 20.
This study aimed to provide further information on the exact mechanisms involved in the anti-inflammatory effect of low-intensity pulsed ultrasound (LIPUS) on rabbit temporomandibular joint osteoarthritis (TMJOA) on interleukin-6 (IL-6) production in subchondral bone, IL-6 production in IL-1β stimulated via inhibition of the TGF-β1/Smad3 pathway in mouse embryo osteoblast precursor (MC3T3-E1) cells.
Bilateral joints were injected with type II collagenase to establish TMJOA models in two male and four female rabbits. The left joint was continuously stimulated by LIPUS, while the right joint was treated with the power off in this model. One male and two female rabbits were used as normal healthy controls without treatment. The histological features of subchondral bone were examined by Safranin-O/Fast staining. Immunohistochemistry was conducted to evaluate IL-6 expression. Then, cells were stimulated by LIPUS with IL-1β. IL-6 expression and activity of the TGF-β1/Smad3 pathway were evaluated by Enzyme-linked immunosorbent assay (ELISA), Immunofluorescence and Western blotting, respectively. Specific inhibition of the TGF-β1/Smad3 pathway was conducted by transfecting with small interfering RNA (siRNA) of type II receptor (siTβRII).
LIPUS significantly ameliorated the production of IL-6 in vitro and in vivo. Its inhibitory effect on the production of IL-6 induced by IL-1β in MC3T3-E1 cells was partly reversed by siTβRII knockdown.
LIPUS inhibited IL-6 production by suppressing the TGF-β1/Smad3 pathway of subchondral bone in TMJOA. These data revealed the part of the pathways involved in the anti-inflammatory effect of LIPUS and provided a possible treatment strategy for TMJOA patients and other inflammatory diseases.
本研究旨在提供关于低强度脉冲超声（LIPUS）对兔颞下颌关节骨关节炎（TMJOA）抑制 TGF-β1/Smad3 通路在 IL-1β刺激下的软骨下骨中白细胞介素-6（IL-6）产生、IL-6 产生的抗炎作用的确切机制的进一步信息在鼠胚胎成骨前体细胞（MC3T3-E1）细胞中。
向双侧关节注射 II 型胶原酶建立 TMJOA 模型，雄性兔 2 只，雌性兔 4 只。左侧关节持续接受 LIPUS 刺激，而右侧关节在该模型中关闭电源。雄性和雌性兔各 1 只作为未治疗的正常健康对照。用番红 O/快染法观察软骨下骨的组织学特征。通过免疫组化评估 IL-6 表达。然后，用 LIPUS 刺激细胞与 IL-1β 共孵育。通过酶联免疫吸附试验（ELISA）、免疫荧光和 Western blot 分别评估 TGF-β1/Smad3 通路的 IL-6 表达和活性。通过转染 II 型受体的小干扰 RNA（siTβRII）对 TGF-β1/Smad3 通路进行特异性抑制。
LIPUS 显著改善了体外和体内的 IL-6 产生。其对 MC3T3-E1 细胞中由 IL-1β诱导的 IL-6 产生的抑制作用部分被 siTβRII 敲低逆转。
LIPUS 通过抑制 TMJOA 软骨下骨 TGF-β1/Smad3 通路抑制 IL-6 产生。这些数据揭示了 LIPUS 抗炎作用涉及的部分途径，并为 TMJOA 患者和其他炎症性疾病提供了一种可能的治疗策略。
