CAJ | 학술논문

目的：探讨河蚌肉提取物葡聚糖HBP-A（anodonta glucan HBP-A）对体外软骨细胞Wnt通路的调控作用。方法：体外培养大鼠软骨细胞，添加IL-1β（10 ng/ml）诱导分化，分为空白组，IL-1β组，IL-1β＋IWP-2（5μM，Wnt通路抑制剂）组，IL-1β＋HBP-A（0．3 mg/ml）组，IL-1β＋IWP-2＋HBP-A共5组培养，提取各组细胞RNA和蛋白，采用Rt-PCR 检测各组细胞 Wnt-3a、β-catenin （24、48、72 h）及 MMP-13（72 h）的基因表达；采用 Western-blot 检测β-catenin、MMP-13、Sox-9和Coll-Ⅱ蛋白的表达水平（48 h）。结果：细胞经IL-1β诱导分化，Wnt-3a基因表达升高，β-catenin以及MMP-13基因和蛋白表达升高，Sox-9和Coll-Ⅱ蛋白表达下调。添加HBP-A干预可以抑制IL-1β诱导下软骨细胞Wnt-3a基因的高表达、β-catenin以及MMP-13基因和蛋白的高表达，上调Sox-9和Ⅱ型胶原蛋白的表达。IWP-2和HBP-A合用时，Wnt-3a、β-catenin基因以及β-catenin蛋白表达最低，Sox-9蛋白表达最高。结论：HBP-A可通过降低Wnt/β-catenin信号通路表达，延缓软骨细胞分化，并且可与Wnt抑制剂协同调节Wnt-3a、β-catenin和Sox-9因子的表达。
목적：탐토하방육제취물포취당HBP-A（anodonta glucan HBP-A）대체외연골세포Wnt통로적조공작용。방법：체외배양대서연골세포，첨가IL-1β（10 ng/ml）유도분화，분위공백조，IL-1β조，IL-1β＋IWP-2（5μM，Wnt통로억제제）조，IL-1β＋HBP-A（0．3 mg/ml）조，IL-1β＋IWP-2＋HBP-A공5조배양，제취각조세포RNA화단백，채용Rt-PCR 검측각조세포 Wnt-3a、β-catenin （24、48、72 h）급 MMP-13（72 h）적기인표체；채용 Western-blot 검측β-catenin、MMP-13、Sox-9화Coll-Ⅱ단백적표체수평（48 h）。결과：세포경IL-1β유도분화，Wnt-3a기인표체승고，β-catenin이급MMP-13기인화단백표체승고，Sox-9화Coll-Ⅱ단백표체하조。첨가HBP-A간예가이억제IL-1β유도하연골세포Wnt-3a기인적고표체、β-catenin이급MMP-13기인화단백적고표체，상조Sox-9화Ⅱ형효원단백적표체。IWP-2화HBP-A합용시，Wnt-3a、β-catenin기인이급β-catenin단백표체최저，Sox-9단백표체최고。결론：HBP-A가통과강저Wnt/β-catenin신호통로표체，연완연골세포분화，병차가여Wnt억제제협동조절Wnt-3a、β-catenin화Sox-9인자적표체。
Objective:To investigate regulation function of anodonta glucan HBP-A on chondrocytes through Wnt path-way in vitro. Methods:Rat chondrocytes were cultured and differentiated induced with IL-1β (10 ng/ml) in vitro. Chondro-cytes were divided into five groups:IL-1βgroup,IL-1β+IWP-2 (5μM,Wnt pathway inhibitor) group,IL-1β+HBP-A (0.3 mg/ml) group and IL-1β+IWP-2+HBP-A group. Wnt-3a,β-catenin (24 h,48 h,72 h) and MMP-13(72 h) genes expres-sion were detected by Rt-PCR,whileβ-catenin,MMP-13,Sox-9 and coll-Ⅱ (48 h) protein expression were measured by Western-blot. Results:After induction of IL-1β,gene expression of Wnt-3a,β-catenin and MMP-13 were increased,so were the protein expression ofβ-catenin and MMP-13. In contrast,protein expression of Sox-9 and Coll-Ⅱwere declined. Follow-ing addition of HBP-A,Wnt-3a,β-catenin and MMP-13 were shown as induction of IL-1β,but protein expression of Sox-9 and Coll-Ⅱwere upgraded . Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a ,β-catenin gene andβ-catenin protein expression and highest expression of Sox-9 protein. Conclusion:HBP-A could not only delay the differentiation of chondrocytes through downgrading the signal expression of Wnt/β-catenin,but also adjust the expression of Wnt-3a,β-catenin and Sox-9 when combinated with the Wnt inhibitor.