中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2015年
15期
2297-2304
,共8页
张紫机%康焱%杨子波%侯昌禾%黄广鑫%陈蔚深%盛璞义%何爱珊%傅明%廖威明%张志奇
張紫機%康焱%楊子波%侯昌禾%黃廣鑫%陳蔚深%盛璞義%何愛珊%傅明%廖威明%張誌奇
장자궤%강염%양자파%후창화%황엄흠%진위심%성박의%하애산%부명%료위명%장지기
组织构建%软骨组织工程%抵抗素%软骨细胞%趋化因子%CCL3%CCL4%转录调控%C/EBPβ%核因子κB%转录后调控%miRNAs%国家自然科学基金
組織構建%軟骨組織工程%牴抗素%軟骨細胞%趨化因子%CCL3%CCL4%轉錄調控%C/EBPβ%覈因子κB%轉錄後調控%miRNAs%國傢自然科學基金
조직구건%연골조직공정%저항소%연골세포%추화인자%CCL3%CCL4%전록조공%C/EBPβ%핵인자κB%전록후조공%miRNAs%국가자연과학기금
Subject headings:Chondrocytes%Chemotactic Factors%NF-kappa B
背景：既往研究表明抵抗素可刺激软骨细胞产生大量趋化因子，在炎症性关节病变中具重要作用，但具体作用机制未明。<br> 目的：进一步探讨抵抗素刺激软骨细胞趋化因子CCL3及CCL4基因表达上调的机制。<br> 方法：培养人源性软骨细胞，T/C-28a2细胞及ATDC5细胞，采用qPCR检测抵抗素刺激趋化因子基因的作用，C/EBPβ表达，核因子κB亚型及软骨特异性miRNAs。给予核因子κB抑制剂(IKK-NBD)和C/EBPβ抑制剂(SB303580)，对C/EBPβ及核因子κB的共同调节作用进行检测。在给予抵抗素刺激或无抵抗素刺激时分别进行亚细胞结构定位检测。<br> 结果与结论：①抵抗素可非依赖性上调趋化因子基因表达。②软骨细胞对抵抗素刺激应答具有非严格细胞特异性，通过C/EBPβ抑制剂、核因子κB及一些软骨细胞特异性miRNAs，可对趋化因子基因表达进行联合调控。③一过性核因子κB活性增高可增强C/EBPβ活性，且两个转录因子对趋化因子基因CCL3及CCL4的作用均为非依赖性。
揹景：既往研究錶明牴抗素可刺激軟骨細胞產生大量趨化因子，在炎癥性關節病變中具重要作用，但具體作用機製未明。<br> 目的：進一步探討牴抗素刺激軟骨細胞趨化因子CCL3及CCL4基因錶達上調的機製。<br> 方法：培養人源性軟骨細胞，T/C-28a2細胞及ATDC5細胞，採用qPCR檢測牴抗素刺激趨化因子基因的作用，C/EBPβ錶達，覈因子κB亞型及軟骨特異性miRNAs。給予覈因子κB抑製劑(IKK-NBD)和C/EBPβ抑製劑(SB303580)，對C/EBPβ及覈因子κB的共同調節作用進行檢測。在給予牴抗素刺激或無牴抗素刺激時分彆進行亞細胞結構定位檢測。<br> 結果與結論：①牴抗素可非依賴性上調趨化因子基因錶達。②軟骨細胞對牴抗素刺激應答具有非嚴格細胞特異性，通過C/EBPβ抑製劑、覈因子κB及一些軟骨細胞特異性miRNAs，可對趨化因子基因錶達進行聯閤調控。③一過性覈因子κB活性增高可增彊C/EBPβ活性，且兩箇轉錄因子對趨化因子基因CCL3及CCL4的作用均為非依賴性。
배경：기왕연구표명저항소가자격연골세포산생대량추화인자，재염증성관절병변중구중요작용，단구체작용궤제미명。<br> 목적：진일보탐토저항소자격연골세포추화인자CCL3급CCL4기인표체상조적궤제。<br> 방법：배양인원성연골세포，T/C-28a2세포급ATDC5세포，채용qPCR검측저항소자격추화인자기인적작용，C/EBPβ표체，핵인자κB아형급연골특이성miRNAs。급여핵인자κB억제제(IKK-NBD)화C/EBPβ억제제(SB303580)，대C/EBPβ급핵인자κB적공동조절작용진행검측。재급여저항소자격혹무저항소자격시분별진행아세포결구정위검측。<br> 결과여결론：①저항소가비의뢰성상조추화인자기인표체。②연골세포대저항소자격응답구유비엄격세포특이성，통과C/EBPβ억제제、핵인자κB급일사연골세포특이성miRNAs，가대추화인자기인표체진행연합조공。③일과성핵인자κB활성증고가증강C/EBPβ활성，차량개전록인자대추화인자기인CCL3급CCL4적작용균위비의뢰성。
BACKGROUND:Previous studies have indicated that resistin stimulates a large set of chemokines in chondrocytes that are known to be important in inflammatory joint lesions. <br> OBJECTIVE:To further investigate the mechanism of co-regulation roles of transcription and post-transcription in the up-regulation of two chemokine genes CCL3 and CCL4 in chondrocytes in response to resistin. <br> METHODS:Human chondrocytes, T/C-28a2 and ATDC5 cels were cultured. The function of resistin on the chemokine genes, and the expression of C/EBPβ, nuclear factor-κB isoforms and chondrogenic specific miRNAs were tested by qPCR. The co-regulation of C/EBPβ and nuclear factor-κB was investigated by nuclear factor-κB inhibitor (IKK-NBD) and C/EBPβ inhibitor (SB303580) treatments, and subcelular localization was detected with or without resistin stimulation. <br> RESULTS AND CONCLUSION:Resistin could increase the expression of chemokine genes independently. Chondrocytes reacted in a non-restrictedly cel-specific manner to resistin; C/EBPβ inhibitor, nuclear factor-κB and some chondrogenic specific miRNAs in a combinatorial manner regulated chemokine gene expression. The activity of C/EBPβ was augmented by a transient increase in activity of nuclear factor-κB, and both transcription factors acted independently on the chemokine genes, CCL3 and CCL4.
