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医学免费论文:黄芩苷对人牙周膜细胞RANKL OPG表达的影响及其作用机制
【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(RANKLOPG)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(TGFβⅡR)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TGFβⅡR基因沉默，继而将转染与未转染靶向性TGFβⅡR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，RTPCR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TGFβⅡR mRNA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TGFβⅡR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;TGFβ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TGFβ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进行调控医.学.全.在.线www.lindalemus.com。
【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
Effect of baicalin on the expression of receptor activator of nuclear factorκB
ligand and osteoprotegerin in human periodontal ligament cellsCHEN Yue1, WU Zhifen2, YANG Lianjia3
(1. Department of Percodontology and Oral Medicine, Oral Hospital, Medical School of
Xian Jiaotong University, Xian 710004; 2. Department of Periodontology and Oral Medicine,
School of Stomatology, Fourth Military Medical University; 3. Department of Oral Histopathology,
School of Stomatology, Fourth Military Medical University, Xian 710032, China)ABSTRACT: Objective To study the effect of baicalin on the expression of receptor activator of nuclear factorκB 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 (TGFβRⅡ), 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 transcriptasepolymerase chain reaction (RTPCR) was used to observe the effect of baicalin on OPGRANKL expression in HPDL cells. Results The clone sequence correctly identified by RTPCR was consistent with the designed target sequence. The recombinant vector was constructed successfully and the expression of TGFβⅡR 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 TGFβⅡR has been constructed and it has been transfected T cells successfully; ② Baicalin can reduce the ratio of RANKL/OPG on PDL cells; ③ TGFβ 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 β to regulate RANKL/OPG in PDL cells, but also through other pathways.
