脐带间充质干细胞对小胶质细胞增殖及活化的影响

doi:10.6040/j.issn.1671-7554.0.2015.1029

摘要/Abstract

摘要： 目的观察人脐带来源的间充质干细胞(hUC-MSCs)在正常环境和炎症环境下对小胶质细胞活化的调节作用,探讨hUC-MSCs对中枢神经系统炎症反应的免疫调节作用。方法将BV2细胞与人脐带间充质干细胞来源的条件培养基(hUC-MSCs-CM)和(或)脂多糖(LPS)共培养24或48 h后利用MTT比色法分析BV2增殖活力的变化,ELISA试剂盒分析BV2细胞上清中TNF-α和IL-6的变化,免疫印迹法检测精氨酸酶1(Arg1)的表达, Real-time PCR分析TNF-α、IL-6、ARG1基因的表达水平。结果 (1)BV2在LPS长程刺激下表现出明显的增殖效应,而hUC-MSCs-CM可以抑制这种增殖反应;(2)炎症刺激诱导BV2表达TNF-α和IL-6明显增多,而在hUC-MSCs-CM和LPS共刺激时TNF-α和IL-6表达明显低于LPS单刺激组;(3)hUC-MSCs-CM共培养组BV2细胞高表达M2型小胶质细胞标志物Arg1。结论脐带间充质干细胞抑制炎症所致的小胶质细胞增殖;脐带间充质干细胞通过旁分泌机制调节小胶质细胞向M2型活化,降低促炎因子表达,这可能是其最终发挥神经保护作用的机制之一。

关键词: 神经保护作用, 脐带间充质干细胞, 免疫调节, 小胶质细胞

Abstract: Objective To observe the effect of umbilical cord mesenchymal stem cells(hUC-MSCs)on the microglia proliferation and activation in normal or inflammatory conditions, in order to explore their immunomodulation effect on the central nervous system. Methods After BV2 cells were co-cultured with hUC-MSCs-CM and/or lipopolysaccharide(LPS)for 24 or 48 h, the proliferation was analyzed by MTT colorimetric method, TNF-α and IL-6 in the supernatant were measured by ELISA kit, Arginase 1(Arg1)protein and mRNA levels were examined by Western blotting, TNF-α and IL-6 mRNA expression level were tested by real-time PCR. Results (1) After long-time LPS stimulation, BV2 cells obviously proliferated, while hUC-MSCs-CM inhibited the proliferation. (2) BV2 cultured with LPS expressed high levels of TNF-α and IL-6, while in both hUC-MSCs-CM and LPS conditions, the TNF-α and IL-6 levels were lower than in LPS stimulation. (3) The level of Arg1 was increased in BV2 cells co-cultured with hUC-MSCs-CM for 48 h. Conclusions Human UC-MSCs inhibit microglia proliferation caused by inflammatory stimulation. Human UC-MSCs induce microglial cells to M2 polarization and reduce the expression of proinflammatory factors via paracrine mechanism, and this may be the mechanism for its neuroprotective effect.

Key words: Neuroprotection, Immunomodulation, Microglia, Umbilical cord mesenchymal stem cells

中图分类号:

R392.1

王娜,陈乃耀. 脐带间充质干细胞对小胶质细胞增殖及活化的影响[J]. 山东大学学报（医学版）, 2016, 54(10): 16-20.

WANG Na, CHEN Naiyao. Effect of umbilical cord mesenchymal stem cells on microglial polarization and proliferation[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(10): 16-20.

参考文献

[1] Saijo K, Glass CK. Microglial cell origin and phenotypes in health and disease[J]. Nat Rev Immunol, 2011, 11(11): 775-787.
[2] Lee WH, Sonntag WE, Mitschelen M, et al. Irradiation induces regionally specific alterations in pro-inflammatory environments in rat brain[J]. Int J Radiat Biol, 2010, 86(2): 132-144.
[3] Jenrow KA, Brown SL, Lapanowski K, et al. Selective inhibition of microglia-mediated neuroinflammation mitigates radiation-induced cognitive impairment[J]. Radiat Res, 2013, 179(5): 549-556.
[4] Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation[J]. Nat Rev Immunol, 2008, 8(12): 958-969.
[5] Chhor V, Le Charpentier T, Lebon S, et al. Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia in vitro[J]. Brain Behav Immun, 2013, 32:70-85. doi: 10.1016/j.bbi.2013.02.005.
[6] Tang Y, Le W. Differential roles of M1 and M2 microglia in neurodegenerative diseases[J]. Mol Neurobiol, 2016, 53(2): 1181-1194.
[7] Hegyi B, Kornyei Z, Ferenczi S, et al. Regulation of mouse microglia activation and effector functions by bone marrow-derived mesenchymal stem cells[J]. Stem Cells Dev, 2014, 23(21): 2600-2612.
[8] Beltrami AP, Cesselli D, Bergamin N, et al. Multipotent cells can be generated in vitro from several adult human organs(heart, liver, and bone marrow)[J]. Blood, 2007, 110(9): 3438-3446.
[9] Gao S, Mao F, Zhang B, et al. Mouse bone marrow-derived mesenchymal stem cells induce macrophage M2 polarization through the nuclear factor-kappaB and signal transducer and activator of transcription 3 pathways[J]. Exp Biol Med(Maywood), 2014, 239(3): 366-375.
[10] Ennis WJ, Sui A, Bartholomew A. Stem cells and healing: impact on inflammation[J]. Adv Wound Care(New Rochelle), 2013, 2(7): 369-378.
[11] Kharraz Y, Guerra J, Mann CJ, et al. Macrophage plasticity and the role of inflammation in skeletal muscle repair[J]. Mediators Inflamm, 2013: 491497. doi: 10.1155/2013/491497.
[12] Mandal P, Pratt BT, Barnes M, et al. Molecular mechanism for adiponectin-dependent M2 macrophage polarization: Link between the metabolic and innate immune activity of full-length adiponectin[J]. J Biol Chem, 2011, 286(15): 13460-13469.
[13] Busch SA, Silver J. The role of extracellular matrix in CNS regeneration[J]. Curr Opin Neurobiol, 2007, 17(1): 120-127.
[14] Tang Y, Li T, Li J, et al. Jmjd3 is essential for the epigenetic modulation of microglia phenotypes in the immune pathogenesis of Parkinsons disease[J]. Cell Death Differ, 2014, 21(3): 369-380.
[15] Rath M, Muller I, Kropf P, et al. Metabolism via Arginase or Nitric Oxide Synthase: Two Competing Arginine Pathways in Macrophages[J]. Front Immunol, 2014, 5: 532. doi: 10.3389/fimmu.2014.00532

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