【摘要】：目的:采用OCILRP2胞外段蛋白OCILRP2-Fc阻斷OCILRP2受體,探討OCILRP2對小鼠樹突狀細胞發(fā)育成熟的影響及其機制。方法:分離培養(yǎng)小鼠骨髓來源的樹突狀細胞(BMDC),熒光定量RT-PCR和流式細胞術分析OCILRP2在不成熟BMDC和LPS誘導的成熟BMDC的表達差異;流式細胞術分析OCILRP2-Fc對BMDC細胞表面標志分子的表達以及對BMDC抗原攝取能力的影響;ELISA檢測培養(yǎng)上清中細胞因子的濃度,分析OCILRP2-Fc對BMDC分泌炎性細胞因子的影響;Western blot檢測轉(zhuǎn)錄因子NF-κB的活化,分析OCILRP2-Fc影響B(tài)MDC分化成熟的分子機制。結(jié)果:熒光定量RT-PCR和流式細胞術結(jié)果均表明LPS可以顯著上調(diào)OCILRP2在BMDC的表達(P0.05);與對照組相比,OCILRP2-Fc可以明顯抑制MHCⅡ類分子及共刺激分子CD80在LPS誘導的成熟BMDC的表達;和成熟BMDC相比,OCILRP2-Fc組BMDC經(jīng)LPS誘導24 h后仍然具有較強的抗原吞噬能力;而且,OCILRP2-Fc組BMDC培養(yǎng)上清中炎性細胞因子IL-6、IL-12、TNF-α的濃度明顯低于對照組(P0.05)。Western blot結(jié)果表明OCILRP2-Fc抑制了LPS誘導的I-κB降解和NF-κB p65的磷酸化。結(jié)論:OCILRP2-Fc阻斷OCILRP2受體信號,通過抑制LPS誘導的NF-κB活化影響小鼠BMDC分化成熟。提示OCILRP2受體在樹突狀細胞的分化成熟過程中具有促進作用。
[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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