三种成体干细胞对脂多糖诱导RAW264.7细胞炎症状态的影响

目的比较人羊膜上皮细胞（（human amniotic epithelial cell, H-AEC））、羊膜间充质细胞（human amniotic mesenchymal
cell, HA-MSC）和脐带间充质细胞（Umbilical cord mesenchymal stem cells, UC-MSC）分泌的细胞因子对脂多糖刺激巨噬细胞
系RAW264.7炎症状态的影响。方法将LPS刺激的RAW264.7细胞炎症模型作为对照组，比较H-AEC、HA-MSC、UC-MSC和
RAW264.7共培养或条件培养基培养RAW264.7对RAW264.7炎症状态的影响。比较各组RAW264.7细胞的迁移能力；检测各
组细胞分泌一氧化氮（NO）的水平；用实时定量多聚酶链反应（RT-PCR）检测各组细胞经典激活的巨噬细胞（classically
activated macrophage, M1 macrophage）相关的促炎基因如白介素-1β（IL-1β）、肿瘤坏死基因а（TNFа）、一氧化氮合成酶-2
（NOS-2）以及M2 macrophage相关的抑炎基因如精氨酸酶（Arg-1）、甘露糖受体基因CD206、B类清道夫受体CD36）表达情况。
结果（1）H-AEC、HA-MSC、UC-MSC 处理后RAW264.7 的迁移率分别为14.7%±4.5%、9.6%±0.7%、13.0%±0.9%，与对照组
（31.1%±11.0%）相比，3 种细胞的条件培养基处理后RAW264.7 的迁移率均降低，差异具有显著性（P<0.05）；（2）H-AEC、
HA-MSC、UC-MSC共培养后RAW264.7 细胞分泌NO的水平分别为24.26±0.72、44.52±2.51、42.25±0.76 μmol/L，与对照组
（45.65±1.78 μmol/L）相比，H-AEC组细胞分泌的NO有显著性下降（P<0.05）；（3）促炎基因与抑炎基因的表达改变：（A）H-AEC
处理组促炎基因IL-1β、TNFа、NOS-2、INFβ的表达下调，与对照组相比有显著差别；HA-MSC、UC-MSC处理组促炎基因INFβ表
达下调显著，其余基因均上调表达；抑炎相关基因如Arg-1、CD206、CD36均上调；（B）3组细胞干预后抑炎症相关基因Arg-1、
CD206、CD36表达均上调，与对照组有显著差异。结论人羊膜上皮细胞、羊膜间充质细胞和脐带间充质细胞可以促进巨噬细
胞向M2型分化，但其效果和机制存在不同。

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

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.