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. 2017 Apr 20;25(4):273-278.
doi: 10.3760/cma.j.issn.1007-3418.2017.04.008.

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

[Article in Chinese]
Y Hou  1 , X Zhou  1 , W L Cai  1 , C C Guo  2 , Y Han  2
Affiliations

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

[Article in Chinese]
Y Hou et al. Zhonghua Gan Zang Bing Za Zhi. .

Abstract
in English, Chinese

Objective: 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. Methods: 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 P < 0.05 was considered as statistically significant. Results: 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%, t = 5.034, P = 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 (t = 11.57 and 10.57, respectively, P < 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%, t = 3.924, P = 0.0172), along with significantly elevated relative mRNA expression of Arg1 (14.35±1.05 vs 1.00±0.03, t = 21.96, P < 0.05) and CD163 (3.04 ± 0.27 vs 1.00 ± 0.03, t = 13.14, P < 0.05). Conclusion: BM-MSCs can inhibit LPS + IFN-γ-induced polarization to M1 macrophages and promote polarization to M2 macrophages through the release of paracrine factors.

目的： 探索骨髓间充质干细胞（BM-MSC）对骨髓来源巨噬细胞极化的影响，为间充质干细胞治疗肝纤维化提供理论依据。 方法： 分别分离小鼠BM-MSC和骨髓来源巨噬细胞，用脂多糖（LPS）和干扰素γ（IFNγ）或白细胞介素4（IL-4）刺激巨噬细胞向M1或M2型巨噬细胞极化。利用Transwell小室建立BM-MSC和巨噬细胞非接触式共培养体系，通过流式细胞术分析共培养后，M1型巨噬细胞和M2型巨噬细胞比例变化情况，通过荧光定量PCR检测M1型巨噬细胞相关分子肿瘤坏死因子α（TNFα）、白细胞介素23a（IL-23a）和M2型巨噬细胞相关分子精氨酸酶1（Arg1）、CD163表达量的变化。两样本比较采用t检验，P<0.05为差异有统计学意义。 结果： 巨噬细胞+ LPS + IFNγ+ BM-MSC共培养组与巨噬细胞+ LPS+IFNγ组比较，M1型巨噬细胞比例显著降低（62.5%±4.6%对比86.6%±6.9%，t = 5.034，P = 0.0073），差异有统计学意义；且M1型巨噬细胞相关分子TNFα相对表达量（3.75±0.23对比6.17±0.27）和IL-23a相对表达量（2.05±0.16对比4.56±0.37），均明显降低，t值分别为11.57、10.57，P值均< 0.05，差异均具有统计学意义。巨噬细胞+ BM-MSC共培养组与巨噬细胞单独培养组相比，M2型巨噬细胞比例显著增高（89.5%±5.8%对比70.1%±6.3%，t = 3.924，P = 0.0172），差异有统计学意义；且M2型巨噬细胞相关分子Arg1相对表达量（14.35±1.05对比1.00±0.03）和CD163相对表达量（3.04±0.27对比1.00±0.03）均显著增高，t值分别为21.96、13.14，P值均< 0.05，差异均具有统计学意义。 结论： BM-MSC可通过旁分泌作用抑制巨噬细胞由LPS + IFNγ诱导的M1型极化，促进巨噬细胞向M2型极化。.

Keywords: Bone marrow mesenchymal stem cells; Liver cirrhosis; Macrophage polarization.

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