人脐带间充质干细胞对巨噬细胞表型调节的机制探讨  

Effect of human umbilical cord-derived mesenchymal stem cells on macrophages phenotype transition

作　　者：薛婧 高杰清[1] 尹雅琪[1] 张琪[1] 于松岩 邹俊彦 郝好杰[2] 母义明[1] XUE Jing1, GAO Jieqing1, YIN Yaqi1, ZHANG Qi1, YU Songyan1, ZOU Junyan1, HAO Haojie2, MU Yiming1(1.Department of Endocrinology; 2Institute of Basic Medicine Chinese PLA General Hospital, Beijing 100853, Chin)

机构地区：[1]解放军总医院内分泌科,北京100853 [2]解放军总医院基础医学所,北京100853

出　　处：《解放军医学院学报》2018年第3期228-233,238,共7页Academic Journal of Chinese PLA Medical School

摘　　要：目的探讨人脐带间充质干细胞(human umbilical cord-derived mesenchymal stem cells,UC-MSCs)对巨噬细胞表型的影响及其机制。方法提取小鼠原代巨噬细胞,分为对照组、脂多糖(lipopolysaccharide,LPS)和干扰素γ(interferon-γ,IFN-γ)诱导组、干细胞共培养组。诱导组加入LPS和IFN-γ诱导24 h,干细胞共培养组在LPS和IFN-γ诱导24 h后与人脐带间充质干细胞通过Transwell共培养24 h。采用q RT-PCR和流式细胞术检测巨噬细胞炎性因子的表达,免疫荧光观察巨噬细胞表型,Western blot检测巨噬细胞极化状态和极化通路关键蛋白的表达。结果干细胞共培养后的巨噬细胞形态改变,长梭形细胞减少,细胞上清IL-1β、TNF-α与诱导组相比分别下降79.5%、41.1%(P均<0.05),抑炎因子IL-4增加91.4%(P<0.05);免疫荧光显示,干细胞共培养组M1型巨噬细胞标记物i NOS阳性细胞比例下降32.48%,M2型巨噬细胞标记物Arg1阳性细胞比例增加32.25%(P均<0.05);Western blot结果显示,参与巨噬细胞极化的PI3K-AKT通路表达增强,而抑制该通路后,巨噬细胞向M2表型极化减弱。结论人脐带来源的间充质干细胞可促进巨噬细胞向M2表型极化,抑制炎症进展,其与PI3K-AKT通路的激活有关。Objective To investigate the effect of human umbilical cord-derived mesenchymal stem cells （UC-MSCs） on macrophages phenotype transition and the potential mechanisms. Methods Macrophages were achieved from C57BL/6J mice and divided into three groups： control group, LPS and IFN- γ induced group and MSCs group. After treated with lipopolysaccharide （LPS） and interferon γ （IFN-γ） for 24 hours, macrophages were co-cultured with UC-MSCs in a Transwell system （MSCs group） or not （LPS and IFN- γ induced group）. The expression of inflammatory cytokines was measured by qRT-PCR and flow cytometry. Macrophages phenotype was detected by immunofluorescence. The polarization state and PI3K/AKT signaling pathway were detected by Western blot. Results After co-cultured with MSCs, macrophages manifested morphological changes with decrease of elongated cells, and levels of IL-1 β, TNF- α in macrophages media reduced by 79.5% and 41.1%, respectively （P 〈 0.05, respectively）, while levels of IL-4 increased by 91.4% compared with those of induced group （P 〈 0.05）. Immunofluorescence analysis revealed that proportion of iNOS positive cells decreased by 32.48% and proportion of Argl positives cells increased by 32.25% in MSCs group （P 〈 0.05, respectively）. Western blot analysis further showed unregulated expression of PI3K and p-AKT in MSCs group, and macrophages polarization towards M2 type was partly blocked after inhibiting PDK/AKT signaling pathway. Conclusion UC-MSCs can polarize macrophages into M2 type and alleviate chronic inflammation, which is related to activation of PI3K-AKT signaling pathway.

关 键 词：间充质干细胞 巨噬细胞 炎症 

分 类 号：R392.12[医药卫生—免疫学]