核心蛋白聚糖协同白细胞介素24对人外周血单个核细胞作用的体外研究

摘要

Objective To investigate the co-effect of decorin (DCN) and interleukin-24 (IL-24) on proliferation and Interferon-γ (IFN-γ) secretion of human peripheral blood mononuclear cells (PBMCs). Methods Recombinant plasmid pcDNA3. 1 (+)-DCN and pcDNA3. 1 (+)-IL-24 were constructed and transfected into PBMCs by liposome. After transfected with different plasmids, the PBMCs were divided into 6 groups;blank control group, Lipofectamine TM group, empty vector group, DCN group, IL-24 group, DCN and IL-24 group. PBMC proliferation was determined by methyl thiazolyl tetrazolium assay. IFN-7 expression in the supernatant was detected by ELISA. Flow cytometry was used to determine the surface expression of programmed death-1 (PD-1) on PBMCs. Statistical analysis was made using LSD-t test. Results The recombinant plasmid pcDNA3. 1 (+)-IL-24 was constructed successfully. PBMC proliferation and IFN-7 secretion were significantly higher in DCN and IL-24 group, and the expression of PD-1 was also upregulated obviously. Conclusion The combination of DCN and IL-24 could promote PBMC proliferation and strengthen the function of PBMCs in vitro.%目的 探讨核心蛋白聚糖(decorin,DCN)联合白细胞介素24(interleukin-24,IL-24)对人外周血单个核细胞(human peripheral blood mononuclear cell,PBMC)增殖及分泌干扰素γ(Interferon-γ,IFN-γ)的影响.方法 构建重组质粒pcDNA3.1(+)-DCN和pcDNA3.1(+)-IL-24,并利用脂质体将两种质粒转染PBMC.根据转染质粒的不同将PBMC分为空白对照组、脂质体组、空质粒组、DCN组、IL-24组、DCN与IL-24联合组.采用四甲基偶氮唑蓝比色法检测PBMC的增殖,ELISA测定细胞培养上清中IFN-γ的表达,流式细胞技术检测PBMC表面程序性死亡分子1(PD-1)的表达.采用LSD-t检验进行统计学分析.结果 重组质粒pcDNA3.1(+)-IL-24构建成功.与其他组相比,DCN与IL-24联合能显著提高PBMC增殖率和IFN-γ分泌量,同时也能上调PD-1的表达水平.结论 DCN与IL-24双基因联合在体外能促进PBMC的增殖,并能增强PBMC的功能.