Department of Orthopedics, Wangjing Hospital of Chinese Academy of Chinese Medical Science, Beijing 100102, PR China; Department of Orthopedics, Xiangtan Hospital Affiliated to Nanhua University, Xiangtan 411101, Hunan Province, PR China.
Department of Orthopedics, Wangjing Hospital of Chinese Academy of Chinese Medical Science, Beijing 100102, PR China.
Transpl Immunol. 2024 Oct;86:102083. doi: 10.1016/j.trim.2024.102083. Epub 2024 Jul 10.
Facet joint osteoarthritis (FJOA) is a common lumbar osteoarthritis characterized by degeneration of small joint cartilage. Bushen Huoxue decotion (BSHXD) has good therapeutic effects on OA. Our work aimed to further probe the pharmacological effects of BSHXD-containing serum (BSHXD-CS) on FJOA and define underlying the mechanisms invovled.
To establish a FJOA cell model, primary rat chondrocytes were treated with LPS. The mRNA and protein expressions were assessed using qRT-PCR and western blot, respectively. The secretion levels of pro-inflammatory cytokines were measured by ELISA. Cell viability was determined by CCK8 assay. The global mA level was detected by the kit, and NLRP3 mRNA mA level was determined by Me-RIP assay. The molecular interactions were analyzed by RIP and RNA pull-down assays.
BSHXD-CS treatment relieved LPS-induced cell injury, inflammation, NLRP3 inflammasome and pyroptosis in chondrocytes (all p < 0.05). LPS-induced NLRP3 upregulation in chondrocytes was related to its high mA modification level (p < 0.05). It was also observed that BSHXD-CS reduced LPS-induced mA modification in chondrocytes via repressing STAT3 (all p < 0.05), suggesting BSHXD-CS could repress NLRP3 expression via mA-dependent manner. Moreover, DAA, a mA specific inhibitor, was proved to strengthen the protectively roles of BSHXD-CS on LPS-challenged pytoptosis (all p < 0.05).
BSHXD-CS inhibited NLRP3 inflammasome activation and pyroptosis in chondrocytes to repress OA progression by reducing RNA mA modification.
小关节骨关节炎（FJOA）是一种常见的腰椎骨关节炎，其特征是小关节软骨退化。补肾活血方（BSHXD）对 OA 有很好的治疗作用。我们的工作旨在进一步探讨含 BSHXD 血清（BSHXD-CS）对 FJOA 的药理作用，并确定其涉及的机制。
建立 FJOA 细胞模型，用 LPS 处理原代大鼠软骨细胞。分别用 qRT-PCR 和 Western blot 检测 mRNA 和蛋白表达。用 ELISA 法测定促炎细胞因子的分泌水平。用 CCK8 法测定细胞活力。用试剂盒检测总 mA 水平，用 Me-RIP 测定 NLRP3 mRNA mA 水平。用 RIP 和 RNA 下拉实验分析分子相互作用。
BSHXD-CS 治疗缓解了 LPS 诱导的软骨细胞损伤、炎症、NLRP3 炎性体和细胞焦亡（均 p<0.05）。LPS 诱导软骨细胞中 NLRP3 上调与它的高 mA 修饰水平有关（p<0.05）。还观察到 BSHXD-CS 通过抑制 STAT3 减少 LPS 诱导的软骨细胞中的 mA 修饰（均 p<0.05），表明 BSHXD-CS 可以通过 mA 依赖性方式抑制 NLRP3 表达。此外，mA 特异性抑制剂 DAA 证实可增强 BSHXD-CS 对 LPS 诱导的细胞焦亡的保护作用（均 p<0.05）。
BSHXD-CS 通过减少 RNA mA 修饰抑制软骨细胞中 NLRP3 炎性体的激活和细胞焦亡，从而抑制 OA 的进展。
