Objective To explore the inhibitory effect of pre-treatment of one marrow mesenchymal stem cells (BMMSCs) with interferon-γ and IFN-γ on the activation of microglia(MG) induced by lipopolysaccharide(LPS).Methods BMMSCs were isolated and cultured using the whole bone marrow adherence method. Then, 100ng/ml IFN-γ was used to induce the activation of BMMSCs and10μg/ml lipopolysaccharide was used as the inflammatory stimulating factor to induce the activation of microglia. After isolation and purification, microglia were seeded in 6-well plates at the density of 1×105/well and BMMSCs were co-cultured with microglia with the cell contact cell method for 24 h in the following groups: B group(BMMSCs group), L+B group(LPS+BMMSCs group), I+B group(IFN-γ pretreatment BMMSCs group), L+I+B group(LPS+IFN-γ pretreatment BMMSCs group), M group(microglia group), L+M group(LPS+ microglia group), B+L+M group(BMMSCs+ LPS+ microglia group), and I+B+L+M group(BMMSCs pretreated with IFN-γ+LPS+ microglia group). 24 hours later, the expressions of TNF-α、IL-1βin cell culture supernatants were detected by ELISA kit and the expression of microglia surface marker CD68 was detected by immunofluorescence.Results In the L+M group,LPS promoted the expression of CD68 and the production of TNF-α and IL-1β in microglial cells. The expression of CD68 and the production of TNF-α and IL-1β in B+L+M group and I+B+L+M group were lower than in L+M group, and there was statistically significant difference between the B+L+M group and the I+B+L+M group(P＜0.05).Conclusions Both BMMSCs and IFN-γ pretreatment of BMMSCs can inhibit the activation of LPS-induced microglia by direct cell contact. After IFN-γ pretreatment ,BMMSCs could more strongly inhibit the activity of microglia.
Key words
bone marrow mesenchymal stem cells /
microglia /
activation /
pretreatment /
IFN-γ
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] Vadivelu S, Willsey M.Potential role of stem cells for neuropathic pain disorders[J].Neurosurgical Focus,2013,35(3):1225-1236.
[2] 王培申,刘学彬,伊 龙.脐带间充质干细胞鞘内移植治疗不完全性颈髓损伤的疗效和安全性[J].武警医学,2015, 26(3):282-285.
[3] 高 锋,张晓峰,段艳伟.经骨髓间充质干细胞诱导分化的神经样细胞对大鼠脊髓损伤的修复作用[J].武警医学,2016, 27(3):285-288.
[4] Prockop D J. Concise review: two negative feedback loops place mesenchymal stem/stromalcells at the center of early regulators of inflammation[J]. Stem Cells,2013,31(10):2042-2046.
[5] Wang Z, Fang B.Hypoxic preconditioning increases the protectiveeffect of bone marrow mesenchymal stem cells on spinal cord ischemia/reperfusion injury[J].Mol Med Rep, 2016,13(3):1953-1960.
[6] MohammadpourH,Pourfathollah A A. Increasing proliferation of murine adipose tissue-derivedmesenchymal stem cells by TNF-α plus IFN-γ[J].Immunopharmaand Immunotoxi,2016;38(2):1-9.
[7] 王炜玮,李 恒. γ干扰素上调人胎盘间充质干细胞PD-L1表达并增强其诱导脐血和外周血T细胞向IL-10+T细胞的分化[J].细胞与分子免疫学杂志, 2016, 32(2):191-195.
[8] Becker A, Riet I V.Homing and migration of mesenchymal stromal cells: how to improve the efficacy of cell therapy[J]. World J Stem,2016, 8(3):73-87.
[9] Wang Z, Fang B, Tan Z, et al. Hypoxic preconditioning increases the protectiveeffectof bone marrow mesenchymal stem cells on spinal cordischemia/reperfusion injury[J].Mol Med Rep,2016,11:1953-1960.
[10] Won Y W, Patel A N, Bull D A, et al. Cell surface engineering to enhance mesenchymal stem cell migration toward an SDF-1 gradient [J]. Biomaterials, 2014, 35(21):5627-5635.
[11] Lan Y W.Hypoxia-preconditioned mesenchymal stem cells attenuates bleomycin-induced pulmonaryfibrosis[J]. Stem Cell Res Ther,2015,6(1):97.
[12] Watanabe S, Nakajima H.Early transplantation of mesenchymal stem cells after spinal cord injury relieves pain hypersensitivitythrough suppression of pain related signaling cascades and reduced inflammatory cell recruitment[J].Stem Cells, 2015, 33(6):1902.
[13] Schfer S, Berger J V, Deumens R.Influence of intrathecal delivery of bone marrow-derived mesenchymal stem cells on spinal inflammation and pain hypersensitivity in a rat model of peripheral nerve injury[J].J Neuroin flammation, 2014, 11(1):157.
[14] Li W, Ren G, Huang Y, et al. Mesenchymal stem cells: a double-edged sword inregulating immune responses[J]. Cell Death Differentiation, 2012,19:1505-1513.
[15] Gazdic M L, Volarevic V. Mesenchymal stem cells: A friend or foe in immune mediated diseases[J]. Stem Cell Rev,2015,11(2):280-287.
[16] Ruth S,Waterman,Suzanne L, et al. A New mesenchymal stem cell (MSC) paradigm: polarization into a pro-Inflammatory MSC1 or an immunosuppressive MSC2 phenotype [J].Plos One, 2010, 5(5):1-14
[17] Gornostaeva A, Andreeva E, Buravkova L.Factors governing the immunosuppressive effects of multipotent mesenchymal stromal cells in vitro [J]. Cytotechnology, 2016, 68(4):1-13.
[18] Sivanathan K N, Gronthos S, Rojas C D,et al. Interferon-gamma modification of mesenchymal stem cells: implications of autologous and allogeneic mesenchymal stem cell therapy in allotransplantation[J]. Stem Cell Rev,2014,10(3):351-375.
[19] Wang L, Zhao Y, Liu Y. IFN-gamma and TNF-alpha synergistically inducemesenchymal stem cell impairment and tumorigenesis via NFkappa B signaling [J]. Stem Cells, 2013,31(7):1383-1395.
[20] 周小琳,罗向阳,何展文.骨髓间充质干细胞对LPS刺激BV2小胶质细胞炎性因子的影响[J].实用医学杂志, 2014,43(22):3545-3548.
PDF(3495 KB)
