醫(yī)學(xué)免費(fèi)論文:黃芩苷對(duì)人牙周膜細(xì)胞RANKL OPG表達(dá)的影響及其作用機(jī)制

醫(yī)學(xué)免費(fèi)論文:黃芩苷對(duì)人牙周膜細(xì)胞RANKL OPG表達(dá)的影響及其作用機(jī)制

【摘要】 目的 研究黃芩苷對(duì)人牙周膜細(xì)胞核因子�拨蔅受體激活物配基和骨保護(hù)素(RANKL�睴PG)表達(dá)的影響及其作用機(jī)制。方法 首先構(gòu)建轉(zhuǎn)化生長(zhǎng)因子βⅡ型受體(TGF�拨垄騌)靶向siRNA真核表達(dá)載體，轉(zhuǎn)染正常人外周血T細(xì)胞，使T細(xì)胞表面的TGF�拨垄騌基因沉默，繼而將轉(zhuǎn)染與未轉(zhuǎn)染靶向性TGF�拨垄騌基因沉默的T細(xì)胞和正常人牙周膜成纖維細(xì)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(xì)胞+內(nèi)毒素+轉(zhuǎn)染siRNA1的T細(xì)胞+黃芩苷;②正常牙周膜成纖維細(xì)胞+內(nèi)毒素+轉(zhuǎn)染siRNA1的T細(xì)胞;③正常牙周膜成纖維細(xì)胞+內(nèi)毒素+正常T細(xì)胞+黃芩苷;④正常牙周膜成纖維細(xì)胞+內(nèi)毒素+正常T細(xì)胞;⑤正常牙周膜成纖維細(xì)胞+黃芩苷;⑥正常牙周膜成纖維細(xì)胞。共培養(yǎng)48h，RT�睵CR檢測(cè)牙周膜細(xì)胞RANKL和OPG的表達(dá)，計(jì)算RANKL/OPG比值。結(jié)果 ①酶切鑒定正確的克隆測(cè)序結(jié)果與設(shè)計(jì)的目的序列完全一致;②轉(zhuǎn)染siRNA1的T細(xì)胞組的TGF�拨垄騌 mRNA的表達(dá)被明顯抑制;③RANKL/OPG的比值在各組間的差異有統(tǒng)計(jì)學(xué)意義(P<0.01)。結(jié)論 成功構(gòu)建了TGF�拨垄騌靶向siRNA真核表達(dá)載體并成功轉(zhuǎn)染T細(xì)胞;黃芩苷可以降低牙周膜細(xì)胞表面RANKL/OPG比值;TGF�拨碌男盘�(hào)傳導(dǎo)在黃芩苷影響牙周膜成纖維細(xì)胞表面的RANKL/OPG表達(dá)過程中起著重要作用;黃芩苷并非只通過TGF�拨滦盘�(hào)傳導(dǎo)通路，而是亦通過其他通路對(duì)牙周膜成纖維細(xì)胞表面的RANKL/OPG進(jìn)行調(diào)控醫(yī).學(xué).全.在.線gydjdsj.org.cn。

【關(guān)鍵詞】 轉(zhuǎn)化生長(zhǎng)因子βⅡ型受體;小干擾RNA;核因子�拨蔅受體激活物配基;骨保護(hù)素;牙周膜成纖維細(xì)胞

Effect of baicalin on the expression of receptor activator of nuclear factor�拨蔅

ligand and osteoprotegerin in human periodontal ligament cellsCHEN Yue1, WU Zhi�瞗en2, YANG Lian�瞛ia3

(1. Department of Percodontology and Oral Medicine, Oral Hospital, Medical School of

Xi�餫n Jiaotong University, Xi�餫n 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, Xi�餫n 710032, China)ABSTRACT: Objective To study the effect of baicalin on the expression of receptor activator of nuclear factor�拨蔅 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�拨翿Ⅱ), 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�瞤olymerase chain reaction (RT�睵CR) was used to observe the effect of baicalin on OPG�睷ANKL expression in HPDL cells. Results The clone sequence correctly identified by RT�睵CR was consistent with the designed target sequence. The recombinant vector was constructed successfully and the expression of TGF�拨垄騌 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�拨垄騌 has been constructed and it has been transfected T cells successfully; ② Baicalin can reduce the ratio of RANKL/OPG on PDL cells; ③ TGF�拨� 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.

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