来源：风湿病与关节炎,2023,12（2）:1-6,14.

乌头汤调控lncRNA GAS5抑制骨关节炎软骨细胞炎症反应的研究（基础研究）

涂海水^1,2，邱志伟^1,2，金灵璐^1,2，郑志煌^1,2，李莹莹^1,2，陈毅涛^1,2，李　超^1,2，付长龙^1,2

　　【摘　要】目的：探讨乌头汤调控lncRNA GAS5进而干预白细胞介素-1β（IL-1β）诱导的软骨细胞炎症反应的效果及机制，为乌头汤的临床应用提供实验依据。方法：4周龄SPF级SD雄性大鼠10只，提取大鼠双膝关节软骨，进行体外软骨细胞的分离与培养，采用Ⅱ型胶原免疫组化法对软骨细胞进行鉴定；经10 ng·mL^-1 IL-1β干预24 h诱导软骨细胞退变。将软骨细胞随机分为空白对照组、模型对照组（10 ng·mL^-1 IL-1β）和乌头汤组（150 μg·mL^-1乌头汤水提物）。通过Real-time PCR检测各组软骨细胞中lncRNA GAS5的相对表达水平。免疫荧光染色技术检测软骨细胞中肿瘤坏死因子-α（TNF-α）、基质金属蛋白酶-3（MMP-3）和解聚蛋白样金属蛋白酶-5（ADAMTS-5）的荧光表达量。结果：与空白对照组比较，模型对照组lncRNA GAS5相对表达水平显著升高（P < 0.01），TNF-α、MMP-3和ADAMTS-5荧光表达增强；与模型对照组比较，乌头汤组lncRNA GAS5相对表达水平显著降低（P < 0.01），TNF-α、MMP-3和ADAMTS-5荧光表达显著减弱。结论：乌头汤可抑制骨关节炎软骨细胞炎症反应，维持软骨细胞外基质稳态，与调控lncRNA GAS5相对水平表达有关。

　　【关键词】　骨关节炎；乌头汤；炎症反应；软骨细胞外基质；大鼠

Study on the Effect of Wutou Tang（乌头汤）on lncRNA GAS5 Inhibiting the Inflammatory Reaction of Chondrocytes in Osteoarthritis

TU Hai-shui,QIU Zhi-wei,JIN Ling-lu,ZHENG Zhi-huang,LI Ying-ying,CHEN Yi-tao,LI Chao,FU Chang-long

　　【ABSTRACT】Objective:To explore the effect and mechanism of chondrocyte inflammatory reaction of Wutou Tang（乌头汤）on the regulation of lncRNA GAS5 and the intervention of interleukin-1β（IL-1β）,provide experimental basis for clinical application of aconite soup.Methods:Ten SPF grade SD male rats at the age of 4 weeks were used to extract the articular cartilage of both knees of the rats and isolate and culture the chondrocytes in vitro.The chondrocytes were identified by typeⅡ collagen immunohistochemistry.After 10 ng·mL^-1 IL-1β intervention for 24 hours induced chondrocyte degeneration.Chondrocytes were randomly divided into blank control group and model control group（10 ng·mL^-1 IL-1β）and aconite soup group（150 μg·mL^-1）.The relative expression level of lncRNA GAS5 in chondrocytes of each group was detected by Real-time PCR.The expression of TNF-α,MMP-3 and ADAMTS-5 in chondrocytes was detected by immunofluorescence staining.Results:Compared with the blank control group,the relative expression level of lncRNA GAS5 in the model control group was significantly higher（P < 0.01）.The fluorescence expression of TNF-α,MMP-3 and ADAMTS-5 was enhanced.Compared with the model control group,the relative expression level of lncRNA GAS5 in aconite decoction group was significantly lower（P < 0.01）.The fluorescence expression of TNF-α,MMP-3 and ADAMTS-5 was significantly decreased.Conclusion:Wutou Tang can inhibit the inflammatory reaction of chondrocytes in osteoarthritis and maintain the stability of extracellular matrix of cartilage,which is related to the regulation of the relative level of expression of lncRNA GAS5.

　　【Keywords】 osteoarthritis;Wutou Tang（乌头汤）;inflammatory reaction;extracellular matrix of cartilage;rats