Department of Orthopedics, The First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital), Nanjing, China.
Physiol Res. 2024 Aug 31;73(4):633-642. doi: 10.33549/physiolres.935284.
The study aims to elucidate the therapeutic mechanism of Baicalin (BAI) in alleviating cartilage injury in osteoarthritic (OA) rat models, concentrating on its regulation of the miR-766-3p/AIFM1 axis. An OA rat model was developed with unilateral anterior cruciate ligament transection (ACLT). Interventions comprised of BAI treatment and intra-articular administration of miR-766-3p inhibitor. For evaluation, histopathological staining was conducted to investigate the pathological severity of knee cartilage injury. The levels of oxidative stress (OS) indicators including MDA, SOD, and GSH-Px, were quantified using colorimetric assays. Inflammatory factors (IFs; TNF-?, IL-1?, and IL-6) in knee joint lavage fluids were assessed using ELISA, while RT-PCR was employed to quantify miR-766-3p expression. TUNEL apoptosis staining was utilized to detect chondrocyte apoptosis, and western blotting examined autophagy-related markers (LC3, Beclin, p62), extracellular matrix (ECM) synthesis-associated indices (COL2A, ACAN, MMP13), and apoptosis-inducing factor mitochondrion-associated 1 (AIFM1). Histological examination revealed a marked amelioration of cartilage injury in the BAI-treated OA rat models compared to controls. BAI treatment significantly reduced inflammation and OS of knee joint fluid, activated autophagy, and decreased chondrocyte apoptosis and ECM degradation. Interestingly, the inhibitory effects of BAI on these pathological markers were significantly decreased by the miR-766-3p inhibitor. Further assessment revealed that BAI efficiently promoted miR-766-3p expression while inhibiting AIFM1 protein expression. BAI potentially mitigates articular cartilage injury in OA rats, likely through modulation of miR-766-3p/AIFM1 axis. Keywords: Baicalin, microRNA, AIFM1, Osteoarthritisv, Rat.
本研究旨在阐明黄芩苷（BAI）通过调节 miR-766-3p/AIFM1 轴缓解骨关节炎（OA）大鼠模型软骨损伤的治疗机制。采用单侧前交叉韧带切断术（ACLT）建立 OA 大鼠模型。干预措施包括 BAI 治疗和关节内注射 miR-766-3p 抑制剂。通过组织病理学染色评估膝关节软骨损伤的病理严重程度。采用比色法测定氧化应激（OS）指标 MDA、SOD 和 GSH-Px 的水平。采用 ELISA 测定关节滑液中炎症因子（TNF-?、IL-1?和 IL-6）的水平，采用 RT-PCR 测定 miR-766-3p 的表达。TUNEL 凋亡染色检测软骨细胞凋亡，Western blot 检测自噬相关标志物（LC3、Beclin、p62）、细胞外基质（ECM）合成相关指标（COL2A、ACAN、MMP13）和凋亡诱导因子线粒体相关 1（AIFM1）。组织学检查显示，与对照组相比，BAI 治疗的 OA 大鼠模型软骨损伤明显改善。BAI 治疗显著降低膝关节液的炎症和 OS，激活自噬，减少软骨细胞凋亡和 ECM 降解。有趣的是，miR-766-3p 抑制剂显著降低了 BAI 对这些病理标志物的抑制作用。进一步评估显示，BAI 可有效促进 miR-766-3p 的表达，同时抑制 AIFM1 蛋白的表达。BAI 可能通过调节 miR-766-3p/AIFM1 轴减轻 OA 大鼠的关节软骨损伤。关键词：黄芩苷，microRNA，AIFM1，骨关节炎，大鼠。
