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[三种不同成体干细胞对脂多糖诱导的RAW264.7细胞炎症状态的影响]

[Effects of three different adult stem cells on inflammatory status of lipopolysaccharide- induced RAW264.7 cells].

作者信息

He Da, Peng Lin, Huang Shengjian, Lu Wenling, Wang Jian

机构信息

Central South University Reproductive And Stem Cell Research Institute, Changsha 410000, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2014 Nov;34(11):1627-31.

Abstract

OBJECTIVE

To compare the modulatory effects of human amniotic epithelial cells (H-AECs), human amniotic mesenchymal cell (HA-MSCs), umbilical mesenchymal cells (UC-MSCs) on the inflammatory status of lipopolysaccharide (LPS)-induced RAW264.7 cells.

METHODS

RAW264.7 cells stimulated with LPS were co-cultured with H-AECs, HA-MSCs, or UC-MSCs or cultured in conditioned media of the 3 stem cells to assess the changes of the inflammatory status of RAW264.7 cells. The migration ability, nitric oxide concentration, and expressions of the pro-inflammatory and anti-inflammatory genes, including interleukin-1β (IL-1β), tumor necrosis factor-α (TNFα), and inducible nitric oxide synthase (NOS-2) of M1 macrophages, and Arg-1, CD206, and CD36 of M2 macrophages, were detected in the co-cultures.

RESULTS

Compared with the control macrophages, RAW264.7 cells cultured in the conditioned media of H-AECs, HA-MSCs, and UC-MSCs all showed significantly lowered migration abilities (P<0.05). Co-culture with H-AECs, but not the other two stem cells, resulted in a significant reduction of NO production (P<0.05) and significant down-regulation of IL-1β, TNFα, NOS-2, and INFβ expressions in RAW264.7 cells; co-culture with HA-MSCs and UC-MSCs only caused a down-regulation of INFβ mRNA expression. In all the 3 RAW264.7 and stem cell co-cultures, the expressions of the inflammation related genes including Arg-1, CD206, and CD36 were up-regulated significantly.

CONCLUSION

H-AECs, HA-MSCs, and UC-MSCs can all prevent RAW264.7 cells from differentiating into M2 macrophages, but their effects and mechanisms are different from one another.

摘要

目的

比较人羊膜上皮细胞(H-AECs)、人羊膜间充质细胞(HA-MSCs)、脐间充质细胞(UC-MSCs)对脂多糖(LPS)诱导的RAW264.7细胞炎症状态的调节作用。

方法

将用LPS刺激的RAW264.7细胞与H-AECs、HA-MSCs或UC-MSCs共培养,或在这3种干细胞的条件培养基中培养,以评估RAW264.7细胞炎症状态的变化。检测共培养物中RAW264.7细胞的迁移能力、一氧化氮浓度以及促炎和抗炎基因的表达,包括M1巨噬细胞的白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNFα)和诱导型一氧化氮合酶(NOS-2),以及M2巨噬细胞的精氨酸酶-1(Arg-1)、CD206和CD36。

结果

与对照巨噬细胞相比,在H-AECs、HA-MSCs和UC-MSCs的条件培养基中培养的RAW264.7细胞迁移能力均显著降低(P<0.05)。与H-AECs共培养可使RAW264.7细胞的一氧化氮产生显著减少(P<0.05),并使IL-1β、TNFα、NOS-2和INFβ的表达显著下调;而与另外两种干细胞共培养仅导致INFβ mRNA表达下调。在所有3种RAW264.7细胞与干细胞的共培养物中,Arg-1、CD206和CD36等炎症相关基因的表达均显著上调。

结论

H-AECs、HA-MSCs和UC-MSCs均可阻止RAW264.7细胞分化为M2巨噬细胞,但其作用及机制各不相同。

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