南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
11期
1627-1631
,共5页
何妲%彭琳%黄生建%陆文玲%王建
何妲%彭琳%黃生建%陸文玲%王建
하달%팽림%황생건%륙문령%왕건
羊膜上皮细胞%羊膜间充质细胞%脐带间充质细胞%M1型巨噬细胞%M2型巨噬细胞
羊膜上皮細胞%羊膜間充質細胞%臍帶間充質細胞%M1型巨噬細胞%M2型巨噬細胞
양막상피세포%양막간충질세포%제대간충질세포%M1형거서세포%M2형거서세포
human amniotic epithelial cells%human amniotic mesenchymal cells%umbilical mesenchymal cells%M1 macrophages%M2 macrophages
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±0.76μmol/L，与对照组（45.65±1.78μmol/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。
目的：比較人羊膜上皮細胞（（human amniotic epithelial cell, H-AEC））、羊膜間充質細胞（human amniotic mesenchymal cell, HA-MSC）和臍帶間充質細胞（Umbilical cord mesenchymal stem cells, UC-MSC）分泌的細胞因子對脂多糖刺激巨噬細胞繫RAW264.7炎癥狀態的影響。方法將LPS刺激的RAW264.7細胞炎癥模型作為對照組，比較H-AEC、HA-MSC、UC-MSC和RAW264.7共培養或條件培養基培養RAW264.7對RAW264.7炎癥狀態的影響。比較各組RAW264.7細胞的遷移能力；檢測各組細胞分泌一氧化氮（NO）的水平；用實時定量多聚酶鏈反應（RT-PCR）檢測各組細胞經典激活的巨噬細胞（classically activated macrophage, M1 macrophage）相關的促炎基因如白介素-1β（IL-1β）、腫瘤壞死基因а（TNFа）、一氧化氮閤成酶-2（NOS-2）以及M2 macrophage相關的抑炎基因如精氨痠酶（Arg-1）、甘露糖受體基因CD206、B類清道伕受體CD36）錶達情況。結果（1）H-AEC、HA-MSC、UC-MSC處理後RAW264.7的遷移率分彆為14.7%±4.5%、9.6%±0.7%、13.0%±0.9%，與對照組（31.1%±11.0%）相比，3種細胞的條件培養基處理後RAW264.7的遷移率均降低，差異具有顯著性（P<0.05）；（2）H-AEC、HA-MSC、UC-MSC共培養後RAW264.7細胞分泌NO的水平分彆為24.26±0.72、44.52±2.51、42.25±0.76μmol/L，與對照組（45.65±1.78μmol/L）相比，H-AEC組細胞分泌的NO有顯著性下降（P<0.05）；（3）促炎基因與抑炎基因的錶達改變：（A）H-AEC處理組促炎基因IL-1β、TNFа、NOS-2、INFβ的錶達下調，與對照組相比有顯著差彆；HA-MSC、UC-MSC處理組促炎基因INFβ錶達下調顯著，其餘基因均上調錶達；抑炎相關基因如Arg-1、CD206、CD36均上調；（B）3組細胞榦預後抑炎癥相關基因Arg-1、CD206、CD36錶達均上調，與對照組有顯著差異。結論人羊膜上皮細胞、羊膜間充質細胞和臍帶間充質細胞可以促進巨噬細胞嚮M2型分化，但其效果和機製存在不同。
목적：비교인양막상피세포（（human amniotic epithelial cell, H-AEC））、양막간충질세포（human amniotic mesenchymal cell, HA-MSC）화제대간충질세포（Umbilical cord mesenchymal stem cells, UC-MSC）분비적세포인자대지다당자격거서세포계RAW264.7염증상태적영향。방법장LPS자격적RAW264.7세포염증모형작위대조조，비교H-AEC、HA-MSC、UC-MSC화RAW264.7공배양혹조건배양기배양RAW264.7대RAW264.7염증상태적영향。비교각조RAW264.7세포적천이능력；검측각조세포분비일양화담（NO）적수평；용실시정량다취매련반응（RT-PCR）검측각조세포경전격활적거서세포（classically activated macrophage, M1 macrophage）상관적촉염기인여백개소-1β（IL-1β）、종류배사기인а（TNFа）、일양화담합성매-2（NOS-2）이급M2 macrophage상관적억염기인여정안산매（Arg-1）、감로당수체기인CD206、B류청도부수체CD36）표체정황。결과（1）H-AEC、HA-MSC、UC-MSC처리후RAW264.7적천이솔분별위14.7%±4.5%、9.6%±0.7%、13.0%±0.9%，여대조조（31.1%±11.0%）상비，3충세포적조건배양기처리후RAW264.7적천이솔균강저，차이구유현저성（P<0.05）；（2）H-AEC、HA-MSC、UC-MSC공배양후RAW264.7세포분비NO적수평분별위24.26±0.72、44.52±2.51、42.25±0.76μmol/L，여대조조（45.65±1.78μmol/L）상비，H-AEC조세포분비적NO유현저성하강（P<0.05）；（3）촉염기인여억염기인적표체개변：（A）H-AEC처리조촉염기인IL-1β、TNFа、NOS-2、INFβ적표체하조，여대조조상비유현저차별；HA-MSC、UC-MSC처리조촉염기인INFβ표체하조현저，기여기인균상조표체；억염상관기인여Arg-1、CD206、CD36균상조；（B）3조세포간예후억염증상관기인Arg-1、CD206、CD36표체균상조，여대조조유현저차이。결론인양막상피세포、양막간충질세포화제대간충질세포가이촉진거서세포향M2형분화，단기효과화궤제존재불동。
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 INFβexpressions 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.
