【摘要】：目的:采用OCILRP2胞外段蛋白OCILRP2-Fc阻斷OCILRP2受體,探討OCILRP2對(duì)小鼠樹突狀細(xì)胞發(fā)育成熟的影響及其機(jī)制。方法:分離培養(yǎng)小鼠骨髓來(lái)源的樹突狀細(xì)胞(BMDC),熒光定量RT-PCR和流式細(xì)胞術(shù)分析OCILRP2在不成熟BMDC和LPS誘導(dǎo)的成熟BMDC的表達(dá)差異;流式細(xì)胞術(shù)分析OCILRP2-Fc對(duì)BMDC細(xì)胞表面標(biāo)志分子的表達(dá)以及對(duì)BMDC抗原攝取能力的影響;ELISA檢測(cè)培養(yǎng)上清中細(xì)胞因子的濃度,分析OCILRP2-Fc對(duì)BMDC分泌炎性細(xì)胞因子的影響;Western blot檢測(cè)轉(zhuǎn)錄因子NF-κB的活化,分析OCILRP2-Fc影響B(tài)MDC分化成熟的分子機(jī)制。結(jié)果:熒光定量RT-PCR和流式細(xì)胞術(shù)結(jié)果均表明LPS可以顯著上調(diào)OCILRP2在BMDC的表達(dá)(P0.05);與對(duì)照組相比,OCILRP2-Fc可以明顯抑制MHCⅡ類分子及共刺激分子CD80在LPS誘導(dǎo)的成熟BMDC的表達(dá);和成熟BMDC相比,OCILRP2-Fc組BMDC經(jīng)LPS誘導(dǎo)24 h后仍然具有較強(qiáng)的抗原吞噬能力;而且,OCILRP2-Fc組BMDC培養(yǎng)上清中炎性細(xì)胞因子IL-6、IL-12、TNF-α的濃度明顯低于對(duì)照組(P0.05)。Western blot結(jié)果表明OCILRP2-Fc抑制了LPS誘導(dǎo)的I-κB降解和NF-κB p65的磷酸化。結(jié)論:OCILRP2-Fc阻斷OCILRP2受體信號(hào),通過(guò)抑制LPS誘導(dǎo)的NF-κB活化影響小鼠BMDC分化成熟。提示OCILRP2受體在樹突狀細(xì)胞的分化成熟過(guò)程中具有促進(jìn)作用。
[Abstract]:Aim: to investigate the effect of OCILRP2 on the development and maturation of mouse dendritic cells by blocking OCILRP2 receptor with OCILRP2 extracellular segment protein OCILRP2-Fc. Methods: (BMDC), quantitative RT-PCR and flow cytometry were used to analyze the difference of OCILRP2 expression in immature BMDC and LPS induced mature BMDC. Flow cytometry was used to analyze the effect of OCILRP2-Fc on the expression of marker molecules on the surface of BMDC cells and on the ability of BMDC antigen uptake, ELISA was used to detect the concentration of cytokines in culture supernatant, and the effect of OCILRP2-Fc on the secretion of inflammatory cytokines by BMDC was analyzed. The activation of transcription factor NF- 魏 B was detected by Western blot, and the molecular mechanism of OCILRP2-Fc affecting the differentiation and maturation of BMDC was analyzed. Results: fluorescence quantitative RT-PCR and flow cytometry showed that LPS could significantly up-regulate the expression of OCILRP2 in BMDC (P0.05). Compared with the control group, OCILRP2-Fc could significantly inhibit the expression of MHC class 鈪，

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