河蚌提取物葡聚糖对软骨细胞Wnt通路的调控作用研究  

Regulating effect of anodonta glucan HBP-A on chondrocytes through Wnt pathway

作　　者：韦宋谱[1,2] 丁道芳[1] 王学宗[1] 庞坚[1] 郑昱新[1] 徐勤光[1] 曹月龙[1] 詹红生[1] 

机构地区：[1]上海中医药大学附属曙光医院骨伤科,上海201203 [2]台州恩泽医疗中心(集团)路桥医院中医骨伤科,浙江台州318050

出　　处：《中国骨伤》2014年第6期461-465,共5页China Journal of Orthopaedics and Traumatology

基　　金：国家自然科学基金(编号:81072830);上海市教委创新项目(编号:11YZ64);上海中医药事业发展三年行动计划项目(编号:ZYSNXD-CC-YJXYY)~~

摘　　要：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。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-1 [3 （10 ng/ml） in vitro. Chondrocytes were divided into five groups ： IL- 1 βgroup, IL- 1 β ＋IWP-2 （5 μM, Wnt pathway inhibitor ） group, IL- 1 [3 ＋ HBP-A （0.3 rag/m1 ） group and IL- 1 β ＋IWP-2 ＋HBP-A group. Wnt-3 a, β -catenin （24 h, 48 h, 72 h ） and MMP- 13 （72 h） genes expres- sion were detected by Rt-PCR,while ^-catenin,MMP-13,Sox-9 and col- H （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 of β-catenin and MMP-13. In contrast ,protein expression of Sox-9 and Coll- Ⅱ were declined. Following addition of HBP-A,Wnt-3a,βcatenin and MMP-13 were shown as induction of IL-1β ,but protein expression of Sox-9 and Coll were upgraded. Combining HBP-A with IWP-2 led to the lowest level in Wnt-3a, β-catenin gene and β-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.

关 键 词：组织提取物 软骨细胞 葡聚糖类 基因表达调控 

分 类 号：R392.11[医药卫生—免疫学]