缺血再灌注条件下内皮祖细胞促进单核/巨噬细胞向M1型极化的研究
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宁夏回族自治区重点研发计划项目(2018EBG02006)


The role and mechanism of endothelial progenitor cells to promote mononuclear/macrophage to type M1 polarization under ischemic reperfusion
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    摘要:

    目的:探讨在缺血再灌注条件下内皮祖细胞(endothelial progenitor cell,EPC)促进单核/巨噬细胞向M1型极化的作用及机制。方法:获取健康志愿者的外周血,原代培养EPC和单核/巨噬细胞;体外构建细胞缺血再灌注损伤模型;流式细胞术检测EPC表面细胞间黏附分子-1(intercellular adhesion molecule -1,ICAM-1)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)和E-选择素的表达;ELISA检测上清中ICAM-1、VCAM-1和E-选择素浓度;采用Transwell小室进行EPC和单核/巨噬细胞共培养,流式细胞术检测M1和M2型单核/巨噬细胞比例。结果:缺血再灌注条件下EPC表面表达及其分泌ICAM-1和VCAM-1均没有显著变化,两组之间差异均没有统计学意义;对照组EPC表面E-选择素平均荧光强度为10.89,缺血再灌注组为33.93(t=3.895,P=0.018);对照组EPC上清中E-选择素浓度为3.69 ng/mL,缺血再灌注组为18.17 ng/mL(t=4.568,P=0.010);缺血再灌注条件下EPC能够促进单核/巨噬细胞向M1型极化,对照组M1型单核/巨噬细胞比例为58.83%,缺血再灌注组为81.43%(t=5.394,P=0.006),E-选择素阻断能抑制这种作用(t=5.950,P=0.004);缺血再灌注条件下EPC抑制单核/巨噬细胞向M2型极化,对照组M2型单核/巨噬细胞比例为60.57%,缺血再灌注组为35.30%(t=6.424,P=0.003),E-选择素阻断能抑制这种作用(t=4.179,P=0.014)。结论:在缺血再灌注损伤条件下,EPC能够通过高表达和分泌E-选择素,促进单核/巨噬细胞向M1型极化,为缺血再灌注损伤的治疗提供了新的靶点。

    Abstract:

    Objective:This study aims to explore the role and mechanism of endothelial progenitor cells(EPC) promoting monocyte/macrophage to type M1 polarization under ischemia and reperfusion. Methods:the peripheral blood of healthy volunteers was obtained,the primary EPC and mononuclear/macrophage were cultured. The model of cell ischemia-reperfusion injury was established in vitro,and the expression of intercellular adhesion molecule -1(ICAM-1),vascular cell adhesion molecule -1(VCAM-1)and E- selectin on EPC surface were detected by flow cytometry. ELISA was used to detect the concentration of ICAM-1,VCAM-1 and E- selectin in the supernatant. Co-culture of EPC and mononuclear/macrophages was performed by Transwell chamber,and the ratio of M1 to M2 type monocytes/macrophages was detected by flow cytometry. Results:There was no significant change in the expression and the secretion of ICAM-1 and VCAM-1 of EPC under the ischemic reperfusion condition,and there was no significant difference between the two groups. The average fluorescence intensity of E- selectin on the surface of EPC was 10.89 in the control group and 33.93 in the ischemia reperfusion group(t=3.895,P=0.018). The concentration of E- selectin in the EPC supernatant was 3.69 ng/mL in the control group and 18.17 ng/mL in the ischemia-reperfusion group(t=4.568,P=0.010). Under ischemia reperfusion condition,EPC promoted monocyte/macrophage to M1 type polarization. The proportion of monocyte/macrophage type M1 was 58.83% in control group and was 81.43% in ischemia reperfusion group(t=5.394,P=0.006),E- selectin blockage could inhibit this effect(t=5.950,P=0.004). Under ischemia reperfusion condition,EPC inhibited monocyte/macrophage to M2 type polarization. The ratio of M2 monocyte/macrophage was 60.57% in control group and was 35.30% in ischemia reperfusion group(t=6.424,P=0.003),E- selectin blockage could inhibit this effect(t=4.179,P=0.014). Conclusion:Under the condition of ischemia-reperfusion injury,EPC can promote the monocyte/macrophage to M1 polarization through the high expression and secretion of E- selectin. Our research provides a new target for the treatment of ischemia-reperfusion injury.

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程 华,金梅花,董俭达.缺血再灌注条件下内皮祖细胞促进单核/巨噬细胞向M1型极化的研究[J].南京医科大学学报(自然科学版),2020,(2):180-184

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  • 收稿日期:2019-07-17
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  • 在线发布日期: 2020-03-14
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