 |
 |
醫(yī)學免費論文:黃芩苷對人牙周膜細胞RANKL OPG表達的影響及其作用機制
【摘要】 目的 研究黃芩苷對人牙周膜細胞核因子κB受體激活物配基和骨保護素(RANKLOPG)表達的影響及其作用機制。方法 首先構(gòu)建轉(zhuǎn)化生長因子βⅡ型受體(TGFβⅡR)靶向siRNA真核表達載體，轉(zhuǎn)染正常人外周血T細胞，使T細胞表面的TGFβⅡR基因沉默，繼而將轉(zhuǎn)染與未轉(zhuǎn)染靶向性TGFβⅡR基因沉默的T細胞和正常人牙周膜成纖維細胞置于加有黃芩苷和內(nèi)毒素的培養(yǎng)基中，分為6組：①正常牙周膜成纖維細胞+內(nèi)毒素+轉(zhuǎn)染siRNA1的T細胞+黃芩苷;②正常牙周膜成纖維細胞+內(nèi)毒素+轉(zhuǎn)染siRNA1的T細胞;③正常牙周膜成纖維細胞+內(nèi)毒素+正常T細胞+黃芩苷;④正常牙周膜成纖維細胞+內(nèi)毒素+正常T細胞;⑤正常牙周膜成纖維細胞+黃芩苷;⑥正常牙周膜成纖維細胞。共培養(yǎng)48h，RTPCR檢測牙周膜細胞RANKL和OPG的表達，計算RANKL/OPG比值。結(jié)果 ①酶切鑒定正確的克隆測序結(jié)果與設計的目的序列完全一致;②轉(zhuǎn)染siRNA1的T細胞組的TGFβⅡR mRNA的表達被明顯抑制;③RANKL/OPG的比值在各組間的差異有統(tǒng)計學意義(P<0.01)。結(jié)論 成功構(gòu)建了TGFβⅡR靶向siRNA真核表達載體并成功轉(zhuǎn)染T細胞;黃芩苷可以降低牙周膜細胞表面RANKL/OPG比值;TGFβ的信號傳導在黃芩苷影響牙周膜成纖維細胞表面的RANKL/OPG表達過程中起著重要作用;黃芩苷并非只通過TGFβ信號傳導通路，而是亦通過其他通路對牙周膜成纖維細胞表面的RANKL/OPG進行調(diào)控醫(yī).學.全.在.線m.quanxiangyun.cn。
【關(guān)鍵詞】 轉(zhuǎn)化生長因子βⅡ型受體;小干擾RNA;核因子κB受體激活物配基;骨保護素;牙周膜成纖維細胞
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.
