黄芩苷对人牙周膜细胞RANKL-OPG表达的影响及其作用机制  

Effect of baicalin on the expression of receptor activator of nuclear factor-κB ligand and osteoprotegerin in human periodontal ligament cells

作　　者：陈悦[1] 吴织芬[2] 杨连甲[3] 

机构地区：[1]西安交通大学医学院口腔医院牙周黏膜科,陕西西安710004 [2]第四军医大学口腔医学院牙周黏膜病科 [3]第四军医大学口腔医学院组织病理科,陕西西安710032

出　　处：《西安交通大学学报（医学版）》2009年第4期492-497,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：西安交通大学医学院口腔医院院青年基金项目

摘　　要：目的研究黄芩苷对人牙周膜细胞核因子-κB受体激活物配基和骨保护素(RANKL-OPG)表达的影响及其作用机制。方法首先构建转化生长因子βⅡ型受体(TGF-βⅡR)靶向siRNA真核表达载体,转染正常人外周血T细胞,使T细胞表面的TGF-βⅡR基因沉默,继而将转染与未转染靶向性TGF-βⅡR基因沉默的T细胞和正常人牙周膜成纤维细胞置于加有黄芩苷和内毒素的培养基中,分为6组:①正常牙周膜成纤维细胞+内毒素+转染siRNA1的T细胞+黄芩苷;②正常牙周膜成纤维细胞+内毒素+转染siRNA1的T细胞;③正常牙周膜成纤维细胞+内毒素+正常T细胞+黄芩苷;④正常牙周膜成纤维细胞+内毒素+正常T细胞;⑤正常牙周膜成纤维细胞+黄芩苷;⑥正常牙周膜成纤维细胞。共培养48h,RT-PCR检测牙周膜细胞RANKL和OPG的表达,计算RANKL/OPG比值。结果①酶切鉴定正确的克隆测序结果与设计的目的序列完全一致;②转染siRNA1的T细胞组的TGF-βⅡRmRNA的表达被明显抑制;③RANKL/OPG的比值在各组间的差异有统计学意义(P<0.01)。结论成功构建了TGF-βⅡR靶向siRNA真核表达载体并成功转染T细胞;黄芩苷可以降低牙周膜细胞表面RANKL/OPG比值;TGF-β的信号传导在黄芩苷影响牙周膜成纤维细胞表面的RANKL/OPG表达过程中起着重要作用;黄芩苷并非只通过TGF-β信号传导通路,而是亦通过其他通路对牙周膜成纤维细胞表面的RANKL/OPG进行调控。Objective To study the effect of baicalin on the expression of receptor activator of nuclear factor-kB ligand （RANKL） and osteoprotegerin （OPG） in cultured human periodontal ligament cells （HPDL cells） as well as its action mechanism. Methods We first constructed small interferring RNA （siRNA） eukaryotic expression vector targeted transforming growth factor βⅡ receptor （TGF-βRⅡ）, and then transfected it into T cells. Then HPDL cells together with T cells transfected with siRNA or not were placed in medium that had been added with lipopolysaccharide （LPS） and baicalin. They were divided into six groups and cultured for 48 hours. Reverse transcriptase-polymerase chain reaction （RT-PCR） was used to observe the effect of baicalin on OPG-RANKL expression in HPDL cells. Results The clone sequence correctly identified by RT-PCR was consistent with the designed target sequence. The recombinant vector was constructed successfully and the expression of TGF-βⅡ R of T cells which had been transfected with siRNA1 was inhibited obviously. Ratio of RANKL/OPG in each group differed significantly （P 〈 0.01）. Conclusion ① siRNA eukaryotic expression vector of recombinant targeting gene TGF-βⅡR has been constructed and it has been transfectcd T cells successfully; ② Baicalin can reduce the ratio of RANKL/OPG on PDL cells; ③ TGF-β signaling transduction plays an important role in the effect of baicalin on RANKL/OPG ratio on PDL cells;④ Baicalin acts not only through TGF βto regulate RANKL/OPG in PDL cells, but also through other pathways.

关 键 词：转化生长因子βⅡ型受体 小干扰RNA 核因子-κB受体激活物配基 骨保护素 牙周膜成纤维细胞 

分 类 号：R781.4[医药卫生—口腔医学]