microRNA-155在弓形虫感染调节巨噬细胞极化中的作用

MicroRNA-155 induces macrophage polarization to M1 in Toxoplasma gondii infection

目的探讨弓形虫感染对宿主细胞内microRNA-155(miR-155)表达的影响及其对巨噬细胞极化的作用。方法利用miRNAs基因芯片检测弓形虫感染宿主细胞内miRNAs表达水平,应用实时定量聚合酶链反应技术(qPCR)检测miR-155表达水平。采用脂质体转染法将pEGFP-miR-155导入人巨噬细胞,利用流式细胞术检测转染效率。采用流式细胞术、qPCR、酶联免疫法检测弓形虫感染组、pEGFP-miR-155过表达组以及miR-155抑制组诱导巨噬细胞表面分子CD86及诱导型一氧化氮合酶(i NOS)和白细胞介素(IL)-12表达水平。结果基因芯片和q PCR检测结果显示,随着弓形虫感染时间的延长,miR-155表达量上升;pEGFP-miR-155转染宿主细胞的转染效率可达82.6%。弓形虫感染组和pEGFP-miR-155过表达组CD86表达水平显著高于对照组和miR-155抑制组,iNOS和IL-12基因表达量显著升高。结论弓形虫感染能够通过上调miR-155表达参与驱动人源巨噬细胞向M1型偏移。

Objective To investigate microRNAs differential expression and polarization of human macrophages in Toxoplasma gondii infection. Methods The microRNAs differential expression of human macrophages in T. gondii infection was analyzed by microarray, and further validated by qRT-PCR. pEGFP-miR-155 was transfected into THP-1 cells by Lipofectamine M2000 and the transfection ratio was detected by flow cytometry. Flow cytometry was used to detect CD86 molecular on the' macrophages.([RT-PCR was used to detect iNOS and IL 12 mRNA expression. NO and IL12 expression were then evaluated by using ELISA. Results The miR -155 up-regulated more than 4-fold in T. gondii infected macrophages and enhanced as well as postinfection prolong. pEGFP-miR-155 transfection ratio was 82.6%. Compared to cells cultured with T. gondii, pEGFP-miR-155 and miR-155-inhibitor, T. gondii and pEGFP-miR-155 inducement enhanced expression of CD86. Additionally, iNOS and IL 12 mRNA were enhanced by qRT-PCR (P < 0.05). NO and IL12 expression were increased by ELISA. Conclusion T. gondii infection up-regulates the host miR-155 expression to modulate macrophages polarization to M1.