医学分子生物学杂志2024，Vol.21Issue(4)：300-308,9.DOI:10.3870/j.issn.1672-8009.2024.04.002
基于NF-κB信号通路探讨miR-155调控的靶基因CEBPβ对颈椎病大鼠椎间盘软骨细胞凋亡和炎症反应的影响
Effect of miR-155 Target Gene CEBPβ on Apoptosis and Inflammato-ry Response of Intervertebral Disc Chondrocytes in Rat with Cervical Spondylosis through Regulation of NF-κB Signaling Pathway
摘要
Abstract
Objective To explore the role and potential mechanisms of CCAAT-enhancer-binding protein β(C/EBPβtargeted by microRNA(miR)-155 in apoptosis and inflammatory re-sponse of rat intervertebral disc chondrocytes with cervical spondylosis.Methods Forty adult male SD rats were randomly divided into 4 groups with 10 rats in each group:Sham group,Cervical Spondylosis group,Cervical Spondylosis+C/EBPβ overexpression group,and Cervical Spondylosis+empty vector group.Rat chondrocyte cell line,atdc5 cells,were divided into 6 groups:control group,tumor necrosis factor α(TNF-α)stimulation group,empty vector+TNF-α group,C/EBPβ overexpression+TNF-α group,mimic-NC+C/EBPβ overexpression+TNF-α group,and miR-155 mimic+C/EBPβ overexpression+TNF-α group.The expression of miR-155,apoptosis-re-lated proteins(cleaved-caspase3,Bax,Bcl-2,PPARγ),NF-κB signaling pathway proteins(p-NF-κB P65,NF-κB P65,IκB),pro-inflammatory cytokines(TNF-α,IL-6,IL-1β),and C/EBPβ in rat cartilage tissues and atdc5 cells were detected using qRT-PCR and Western blotting ex-periments.Dual-luciferase reporter gene assay was conducted to determine the targeted regulatory effect of miR-155 on C/EBPβ expression.Results miR-155,cleaved-caspase3,Bax,PPARγ,p-NF-κB P65,TNF-α,IL-6,and IL-1β were upregulated in the Cervical Spondylosis group when compared with those in the Sham group,while Bcl-2,IκB,and C/EBPβ were downregulated in the Cervical Spondylosis group(P<0.05)Ceaved-caspase3,Bax,PPARγ,p-NF-κB P65,TNF-α,IL-6,and IL-1β were downregulated in the Cervical Spondylosis+C/EBPβ overexpression group when compared with those in the Cervical Spondylosis+empty vector group,while Bcl-2,IκB,and C/EBPβ were upregulated in the Cervical Spondylosis+C/EBPβ overexpression group(P<0.05),but there were no significant change in miR-155 expression level(P>0.05).There was a significant negative correlation between the expression levels of miR-155 and C/EBPβ in the cartilage of rats with cervical spondylosis(r=-0.721,P<0.05).The TNF-α group showed up-regulations of miR-155,cleaved-caspase3,Bax,PPARγ,p-NF-κB P65,IL-6,and IL-1β ex-pression(P<0.05),and downregulations of Bcl-2,IκB,and C/EBPβ expression when com-pared with the control group(P<0.05).The C/EBPβ overexpression+TNF-α group showed down-regulations of cleaved-caspase3,Bax,PPARγ,p-NF-κB P65,IL-6,and IL-1β expression,and upregulations of Bcl-2,IκB,and C/EBPβ expression when compared with the empty vector+TNF-α group(P<0.05),but no significant change was observed in miR-155 expression level(P>0.05).The mimic+C/EBPβ overexpression+TNF-α group showed upregulations of miR-155,cleaved-caspase3,Bax,PPARγ,p-NF-κB P65,IL-6,and IL-1β expression(P<0.05),and downregulations of Bcl-2,IκB,and C/EBPβ expression when compared with the mimic-NC+C/EBPβ overexpression+TNF-α group(P<0.05).Dual-luciferase reporter gene assay confirmed CEBPβ as a target gene of miR-155 in chondrocytes.Conclusion miR-155 promotes the activation of the NF-κB signaling pathway in intervertebral disc cartilage tissues and chondrocytes in rat with cervical spondylosis by inhibiting the target gene CEBPβ,which leads to enhanced apoptosis and in-flammatory response.关键词
微小RNA-155/CCAAT增强子结合蛋白β/颈椎病大鼠/软骨/凋亡/炎症反应Key words
microRNA-155/CCAAT-enhancer-binding protein β/rat with cervical spondylo-sis/cartilage/apoptosis/inflammatory response分类
医药卫生引用本文复制引用
纳青,吐尔逊娜依·阿布都热依木,吴刚..基于NF-κB信号通路探讨miR-155调控的靶基因CEBPβ对颈椎病大鼠椎间盘软骨细胞凋亡和炎症反应的影响[J].医学分子生物学杂志,2024,21(4):300-308,9.基金项目
新疆维吾尔自治区自然科学基金(No.2022D01C326) This work was supported by a grant from the Natural Science Foundation of Xinjiang Uygur Autonomous Region(No.2022D01C326) （No.2022D01C326）
