Abstract: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.
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.
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.