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巨噬细胞吞噬骨髓间充值干细胞后对梗死心肌修复作用及可能机制的研究

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DOI:
作者:
陆文彬 傅聪 沈成兴
Wenbin，Lu； Cong，Fu ；Chengxing,Shen
作者单位:
东南大学
Southeast University
关键词:
巨噬细胞；骨髓间充值干细胞；细胞移植；pMΦ
摘要:
摘要： 目的：研究巨噬细胞（MΦ）吞噬骨髓间充值干细胞（MSCs）后对梗死心肌的修复作用以及可能的机制。方法：体外分离培养骨髓间充值干细胞，经流式细胞仪分离纯化及及进一步缺氧诱导凋亡后与LPS激活的巨噬细胞共培养48小时，随后获得吞噬了骨髓间充值干细胞的巨噬细胞（pMΦ）；PBS组为对照，将MSCs、pMΦ、MIP-1α+pMΦ三组细胞经尾静脉注射入急性心肌梗死后BALB/C小鼠体内；磁共振及多光谱分析仪分析移植后细胞在梗死区的定植及强度；小鼠心脏超声及弹力纤维染色分析移植细胞对小鼠梗死心肌的修复作用；流式细胞技术分析移植细胞修复小鼠梗死心肌的可能机制。结果：与PBS组相比，三组细胞均可定植于梗死心肌局部，三治疗组之间两两比较均差异显著（p＜0.001）；心肌梗死后两周小鼠心脏超声示pMΦ组改善心功能优于MSCs组但差于MIP-1α+pMΦ治疗组（p＜0.001）；弹力纤维染色可见pMΦ、MIP-1α+pMΦ两组细胞治疗后较单纯MSCs组弹力纤维增生显著（p＜0.001），但两组之间未见明显差异（p=0.32）；经胰酶消化及流式细胞仪检测可见pMΦ治疗组梗死区局部CD86阳性巨噬细胞显著多于MSCs治疗组，但少于MIP-1α+pMΦ组（p＜0.001）结论：急性心肌梗死后移植吞噬了凋亡MSCs后的巨噬细胞可有效定植于梗死心肌并显著改善梗死后的心脏功能，机制与促进弹力纤维增生及促使更多修复性性巨噬细胞的动员与募集有关，MIP-1α可增强这种作用。
[Abstract] Objective: To study the effect of macrophages (MΦ) after phagocytized died bone marrow stem cells (MSCs) on the repairation of infarcted myocardium and the possible mechanisms. Methods: Bone marrow stem cells were cultured in vitro , after being purified by flow cytometry and further being hypoxia, the dead MSCs were then co-cultured with LPS-activated macrophages for 48 hours, then pMΦ (macrophage which has phagocytized died MSCs) were got; PBS as a control group , MSCs, pMΦ, MIP-1α + pMΦ were injected into acute myocardial infarction BALB / C mice through tail vein; MRI and multi-spectrometer analyzer were used to identify the colonization intensity of the transplanted cells in the infarct area; Mice cardiac ultrasound and elastic fibers stain were used to analyze the effection of the transplanted cells on the repairation of infarcted myocardium; Flow cytometry were being used to speculate the possible mechanisms of transplanted cells on repairation of infarcted myocardium in mice. Results: Compared to PBS group, all the three groups of cells can be planted in myocardial infarction, comparisons between any two groups were significantly (p <0.001); Echocardiography showed improved heart function in pMΦ group better than that of MSCs group but less than MIP-1α + pMΦ group (p <0.001) two weeks after myocardial infarction; Elastic fiber staining showed improved elastic fibers in pMΦ and MIP-1α + pMΦ group compared to MSCs group (p <0.001)but did not show significant difference between the two groups (p = 0.32); After trypsin digestion and flow cytometry detection found that CD86-positive macrophages localized significantly in pMΦ group, more than that of MSCs treated group, but less than MIP-1α + pMΦ group (p <0.001) Conclusion: pMΦ can effectively colonize in the infarcted myocardium after being transplanted into acute myocardial infarction mice and can significantly improved cardiac function, the possible mechanisms may related to the promotion of elastic fibers proliferation and mobilization of more reparative macrophages, MIP-1α can enhance this effect.

 

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