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(150μg·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。
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 typeⅡcollagen immunohistochemistry.After 10 ng·mL^(-1) IL-1βintervention 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(150μg·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 re-lated to the regulation of the relative level of expression of lncRNA GAS5.
作者
涂海水
邱志伟
金灵璐
郑志煌
李莹莹
陈毅涛
李超
付长龙
TU Hai-shui;QIU Zhi-wei;JIN Ling-lu;ZHENG Zhi-huang;LI Ying-ying;CHEN Yi-tao;LI Chao;FU Chang-long
出处
《风湿病与关节炎》
2023年第2期1-6,14,共7页
Rheumatism and Arthritis
基金
福建省自然科学基金项目(2022J01370)
福建中医药大学高层次人才科研启动资金项目(X2019011-人才)
陈可冀中西医结合发展基金资助项目(CKJ2021018)。
关键词
骨关节炎
乌头汤
炎症反应
软骨细胞外基质
大鼠
osteoarthritis
Wutou Tang(乌头汤)
inflammatory reaction
extracellular matrix of cartilage
rats
