从lncRNA NEAT1/miRNA-377-3p调控角度探讨乌头汤改善骨关节炎软骨退变的实验研究

Experimental Study on the Improvement of in Osteoarthritis Cartilage Degenerationby Wutou Tang(乌头汤)from the Perspective of lncRNA NEAT1/miRNA-377-3p Regulation

目的:探讨乌头汤通过调控lncRNA NEAT1/miRNA-377-3p对骨关节炎软骨退变的作用机制。方法:饲养C57BL/6小鼠48只,随机分为空白对照组12只和造模组36只。采用Hulth法制备骨关节炎模型,模型建立后,再随机分为模型对照组、乌头汤组和硫酸氨基葡萄糖组,每组12只,番红-固绿染色进行骨关节炎模型鉴定。待干预完成后,采用Micro-CT观察膝关节形态变化,采用Real-time PCR检测软骨组织中lncRNA NEAT1、miRNA-377-3p、MMP-9、MMP-13、ADAMTS-4、ADAMTS-5、Aggrecan水平变化。结果:Micro-CT结果显示:与空白对照组比较,模型对照组小鼠关节结构变形明显,关节间隙严重狭窄,骨关节面破坏程度高且伴有骨赘形成;与模型对照组比较,乌头汤组、硫酸氨基葡萄糖组小鼠关节变形及损害程度发生显著改善。与空白对照组比较,模型对照组lncRNA NEAT1、MMP-9、MMP-13、ADAMTS-4、ADAMTS-5水平显著增高,miRNA-377-3p、Aggrecan相对表达水平显著降低(P<0.05);而乌头汤组可显著降低lncRNA NEAT1、MMP-9、MMP-13、ADAMTS-4、ADAMTS-5水平,并上调miRNA-377-3p、Aggrecan水平(P<0.05)。结论:乌头汤可改善骨关节炎软骨退变,与调控lncRNA NEAT1/miRNA-377-3p有关。

Objective:To investigate the mechanism of Wutou Tang(乌头汤)in regulating the expression of lncRNA NEAT1/miRNA-377-3p in osteoarthritis cartilage degeneration.Methods:Forty-eight C57BL/6 mice were randomly divided into a blank control group(12 mice)and a model group(36 mice).The Hulth method was used to prepare the osteoarthritis model.After the model was established,it was randomly divided into a model control group,a Wutou Tang group,and a glucosamine sulfate capsule group,with 12 animals in each.The osteoarthritis model was identified using safranin-fast green staining.After the intervention,Micro-CT was used to observe the changes in the morphology of the knee joint,and Real-time PCR was used to detect the changes in the levels of lncRNA NEAT1,miRNA-377-3p,MMP-9,MMP-13,ADAMTS-4,ADAMTS-5,and Aggrecan in cartilage tissues.Results:Micro-CT results show:compared with the blank control group,the joint structure of the model control group was significantly deformed,with severe narrowing of the joint space,high degree of destruction of the bone articular surface,and accompanied by osteophyte formation;compared with the model control group,the degree of joint deformation and damage in mice was significantly improved in the Wutou Tang group and the glucosamine sulfate capsule group.Compared with the blank control group,the levels of lncRNA NEAT1,MMP-9,MMP-13,ADAMTS-4,and ADAMTS-5 in the model control group were significantly increased,while the relative expression levels of miRNA-377-3p and Aggrecan were significantly decreased(P<0.05);in the Wutou Tang group,the levels of lncRNA NEAT1,MMP-9,MMP-13,ADAMTS-4,and ADAMTS-5 were significantly reduced,and the levels of miRNA-377-3p and Aggrecan were upregulated(P<0.05).Conclusion:Wutou Tang can improve cartilage degeneration in osteoarthritis,which is related to the regulation of lncRNA NEAT1/miRNA-377-3p.