人骨髓间充质干细胞过表达白细胞介素-34对THP-1细胞的调节作用

摘要

To construct recombinant eukaryotic expression plasmid vector of human IL-34 gene,and to study the effects of IL-34 expressed by human bone marrow-derived mesenchymal stem cells (hBM-MSCs) on THP-1 cells.Full-length IL-34 encoding sequence was amplified by PCR.And this fragment was cloned into the plasmid pIRES2-EGFP.Western blotting and ELISA were used to analyze the expression of IL-34 in hBM-MSCs.THP-1 cells were cultured with hBM-MSCs medium containing IL-34 protein.Real-time PCR detected the effects of IL-34 on the expression of IL-10 and TNFα in THP-1 cells.Restrictive enzyme analysis and sequencing demonstrated that IL-34 eukaryotic expression vector was successfully constructed.IL-34 protein expressed by hBM-MSCs could promote IL-10 and TNFα expression in THP-1 cells.Those results show that IL-34 expressed by hBM-MSCs has regulating effect on THP-1 cells.%构建人IL-34真核表达载体并将其转染到人骨髓间充质干细胞,观察高表达IL-34的骨髓间充质干细胞对THP-1细胞的影响.PCR扩增IL-34 DNA,并将其克隆到真核表达载体pIRES2-EGFP;将构建成功的重组体转染到骨髓间充质干细胞,Western blotting和ELISA分析IL-34在细胞中的表达;用高表达IL-34的骨髓间充质干细胞培养上清液来培养THP-1细胞,Real-time PCR分析THP-1细胞中IL-10和TNFα的表达变化.经双酶切和测序鉴定,成功构建了pIRES2-EGFP-IL-34重组体;转染至骨髓间充质干细胞的IL-34可以促进THP-1细胞表达IL-10和TNFα.结果表明,骨髓间充质干细胞表达分泌的IL-34对THP-1有调节作用.