miR-655-3p调节DDX3X对IL-1β诱导的骨关节炎软骨细胞损伤的影响OA
Effect of miR-655-3p on IL-1β-Induced Chondrocyte Injury in Osteoarthritis by Regulating DDX3X
目的:探究微小 RNA-655-3p(miR-655-3p)在白细胞介素-1β(IL-1β)诱导的骨关节炎软骨细胞损伤中的作用,解析其与 DEAD-box RNA 解旋酶 3X 连锁(DDX3X)之间存在的关联.方法:用 IL-1β 诱导人软骨细胞系 CHON-001 建立软骨细胞损伤模型.软骨细胞进行随机分组:对照组、模型组、模型+miR-NC 组、模型+miR-655-3p mimics 组、模型+sh-NC 组、模型+sh-DDX3X 组、模型+miR-655-3p mimics+pcDNA3.1 组、模型+miR-655-3p mimics+pcDNA3.1-DDX3X 组.qRT-PCR 检测miR-655-3p 和 DDX3X mRNA 表达;MTT 法检测 CHON-001 细胞增殖;TUNEL 染色检测细胞凋亡;ELISA 法检测 IL-6、TNF-α 和 IL-10 水平;Western blot 法检测 Bcl-2、Bax、DDX3X 蛋白表达;miR-655-3p 和 DDX3X 关系验证用双荧光素酶报告实验.结果:与对照组比较,模型组 miR-655-3p 表达、IL-10水平、Bcl-2 表达降低,DDX3X 表达、细胞增殖抑制率、凋亡率、IL-6、TNF-α 水平、Bax 表达升高(P<0.05);与模型组、模型+miR-NC 组比较,模型+miR-655-3p mimics 组 miR-655-3p 表达、IL-10 水平、Bcl-2 表达升高,DDX3X 表达、细胞增殖抑制率、凋亡率、IL-6、TNF-α 水平、Bax 表达降低(P<0.05);与模型组、模型+sh-NC 组比较,模型+sh-DDX3X 组 miR-655-3p 表达无差异(P>0.05),DDX3X 表达、细胞增殖抑制率、凋亡率、IL-6、TNF-α 水平、Bax 表达降低,IL-10 水平、Bcl-2 表达升高(P<0.05);与模型+miR-655-3p mimics 组、模型+miR-655-3p mimics+pcDNA3.1 组比较,模型+miR-655-3p mimics+pcD-NA3.1-DDX3X 组 miR-655-3p 表达无差异(P>0.05),DDX3X 表达、细胞增殖抑制率、凋亡率、IL-6、TNF-α 水平、Bax 表达升高,IL-10 水平、Bcl-2 表达降低(P<0.05).miR-655-3p 和 DDX3X 存在靶向关系.结论:miR-655-3p 过表达能够抑制 DDX3X 表达,进而改善 IL-1β 诱导的骨关节炎软骨细胞损伤.
Objective:To investigate the role of microRNA-655-3p(miR-655-3p)in interleukin-1β(IL-1β)-induced chondrocyte injury in osteoarthritis,and to analyze its association with DEAD-box RNA helicase 3X-linked(DDX3X).Methods:A chondrocyte injury model was established by inducing human chondrocyte line CHON-001 with IL-1β.Chondrocytes were randomly assigned into control group,model group,model+miR-NC group,model+miR-655-3p mimics group,model+sh-NC group,model+sh-DDX3X group,model+miR-655-3p mimics+pcDNA3.1 group,and model+miR-655-3p mimics+pcDNA3.1-DDX3X group.qRT-PCR was used to detect the expression of miR-655-3p and DDX3X mRNA;MTT assay was used to detect CHON-001 cell proliferation;TUNEL staining was used to detect cell apoptosis;ELISA was used to detect the levels of IL-6,TNF-α,and IL-10;Western blot was used to detect the protein ex-pression of Bcl-2,Bax,and DDX3X;the relationship between miR-655-3p and DDX3X was validated using dual luciferase reporter assay.Results:Compared with the control group,the model group showed decreases in miR-655-3p,IL-10,and Bcl-2,and raises in DDX3X,cell proliferation inhibition rate,apoptosis rate,IL-6,TNF-α,and Bax(P<0.05).Compared with the model group and model+miR-NC group,the model+miR-655-3p mimics group showed raises in miR-655-3p,IL-10,and Bcl-2,and reductions in DDX3X,cell proliferation inhibition rate,apoptosis rate,IL-6,TNF-α,and Bax(P<0.05).Compared with the model group and model+sh-NC group,the model+sh-DDX3X group showed no significant difference in miR-655-3p(P>0.05),but reductions in DDX3X,cell proliferation inhibition rate,apoptosis rate,IL-6,TNF-α,and Bax,and increases in IL-10,and Bcl-2(P<0.05).Compared with the model+miR-655-3p mim-ics group and model+miR-655-3p mimics+pcDNA3.1 group,the model+miR-655-3p mimics+pcDNA3.1-DDX3X group showed no significant difference in miR-655-3p(P>0.05),but increases in DDX3X,cell proliferation inhibition rate,apoptosis rate,IL-6,TNF-α,and Bax,and reductions in IL-10 and Bcl-2(P<0.05).There was a targeting relationship between miR-655-3p and DDX3X.Conclusions:Overexpres-sion of miR-655-3p can inhibit DDX3X,thereby improving IL-1β-induced chondrocyte injury in osteoar-thritis.
刘立冰;祝丰成;李海欣;李焕龙;蔡怡
湖北省武汉市第五医院骨科,湖北 武汉 430001云南省昆明市第二人民医院骨科,云南 昆明 650202云南省昆明市第二人民医院骨科,云南 昆明 650202云南省昆明市第二人民医院骨科,云南 昆明 650202湖北省武汉市第五医院骨科,湖北 武汉 430001
骨关节炎miR-655-3pDDX3XIL-1β软骨细胞
OsteoarthritismiR-655-3pDDX3XIL-1βChondrocyte
《河北医学》 2026 (7)
1084-1090,7
昆明市卫生健康委员会卫生科研课题项目(2025-04-07-005)
评论