骨髓间充质干细胞对巨噬细胞极化的调控作用

[Regulatory effect of bone marrow mesenchymal stem cells on polarization of macrophages].

作者信息

Hou Y, Zhou X, Cai W L, Guo C C, Han Y

机构信息

State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an 710032, China.

Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, Xi'an 710032, China.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2017 Apr 20;25(4):273-278. doi: 10.3760/cma.j.issn.1007-3418.2017.04.008.

DOI:10.3760/cma.j.issn.1007-3418.2017.04.008

PMID:28494546

Abstract

To examine the regulatory effect of bone marrow mesenchymal stem cells (BM-MSCs) on the polarization of bone marrow-derived macrophages, and to provide a theoretical support for the application of mesenchymal stem cells in the treatment of liver fibrosis. MSCs and macrophages were first isolated from the bone marrow of mice. Macrophages were polarized to M1 macrophages with lipopolysaccharide (LPS) and interferon-γ (IFN-γ), and to M2 macrophages with interleukin-4 (IL-4). The macrophages were then co-cultured with BM-MSCs in a Transwell for 24 h, and changes in the percentages of M1 and M2 macrophages were examined using flow cytometry. The mRNA levels of the M1 macrophage-associated cytokines, tumor necrosis factor-α (TNF-α) and interleukin-23a (IL-23a), and M2 macrophage-associated molecules, arginase-1 (Arg-1) and CD163, were measured by real-time quantitative PCR. The two samples were compared using the t test, and < 0.05 was considered as statistically significant. Flow cytometry showed that the percentage of M1 macrophages was significantly lower in the (macrophage + LPS + IFN-γ + BM-MSC) co-culture group than in the (macrophage + LPS + IFN-γ) group (62.5% ± 4.6% vs 86.6% ± 6.9%, = 5.034, = 0.0073). In addition, the relative mRNA expression of TNF-α and IL-23a was also significantly reduced in the co-culture group compared with those in the macrophage control group as measured by RT-qPCR ( = 11.57 and 10.57, respectively, < 0.05). Compared with that in the macrophage control group, the percentage of M2 macrophages in the (macrophage+BM-MSC) co-culture group was significantly increased (89.5% ± 5.8% vs 70.1% ± 6.3%, = 3.924, = 0.0172), along with significantly elevated relative mRNA expression of Arg1 (14.35±1.05 vs 1.00±0.03, = 21.96, < 0.05) and CD163 (3.04 ± 0.27 vs 1.00 ± 0.03, = 13.14, < 0.05). BM-MSCs can inhibit LPS + IFN-γ-induced polarization to M1 macrophages and promote polarization to M2 macrophages through the release of paracrine factors.

摘要

探讨骨髓间充质干细胞（BM-MSCs）对骨髓来源巨噬细胞极化的调控作用，为间充质干细胞在肝纤维化治疗中的应用提供理论支持。首先从小鼠骨髓中分离出间充质干细胞和巨噬细胞。用脂多糖（LPS）和干扰素-γ（IFN-γ）将巨噬细胞极化为M1巨噬细胞，用白细胞介素-4（IL-4）将其极化为M2巨噬细胞。然后将巨噬细胞与BM-MSCs在Transwell中共培养24小时，并用流式细胞术检测M1和M2巨噬细胞百分比的变化。通过实时定量PCR检测M1巨噬细胞相关细胞因子肿瘤坏死因子-α（TNF-α）和白细胞介素-23a（IL-23a）以及M2巨噬细胞相关分子精氨酸酶-1（Arg-1）和CD163的mRNA水平。两组样本采用t检验进行比较，P<0.05被认为具有统计学意义。流式细胞术显示，（巨噬细胞+LPS+IFN-γ+BM-MSC）共培养组中M1巨噬细胞的百分比显著低于（巨噬细胞+LPS+IFN-γ）组（62.5%±4.6%对86.6%±6.9%，t=5.034，P=0.0073）。此外，通过RT-qPCR检测，共培养组中TNF-α和IL-23a的相对mRNA表达也较巨噬细胞对照组显著降低（分别为t=11.57和10.57，P<0.05）。与巨噬细胞对照组相比，（巨噬细胞+BM-MSC）共培养组中M2巨噬细胞的百分比显著增加（89.5%±5.8%对70.1%±6.3%，t=3.924，P=0.0172），同时Arg1（14.35±1.05对1.00±0.03，t=21.96，P<0.05）和CD163（3.04±0.27对1.00±0.03，t=13.14，P<0.05）的相对mRNA表达也显著升高。BM-MSCs可通过旁分泌因子的释放抑制LPS+IFN-γ诱导的向M1巨噬细胞的极化，并促进向M2巨噬细胞的极化。