Combination of G-CSF and AMD3100 Improves the Anti-inflammatory Effect of Mesenchymal Stem Cells on Inducing M2 Polarization of Macrophages Through NF-κB-IL1RA Signaling Pathway.

Long Chen,Feng-Yin Ran,Li-Mei Yu,Gong-Yu Song,Qian Zhang,Tao Zhang,Jun-Ming Tang,Qiang Fu,Xiu Liu,Qin-Hua Chen

doi:10.3389/fphar.2019.00579

Abstract

Mobilized peripheral blood-derived mesenchymal stem cells (PB-MSCs) mainly derived from bone marrow-derived MSCs (BM-MSCs) exert a similar anti-inflammatory effect. However, the mechanism of anti-inflammatory effect of mobilized PB-MSCs by a combination of G-CSF and AMD3100 remains unclear. Cultured rat PB-MSCs mobilized by G-CSF/AMD3100 have shown typical surface markers and potential for multiple differentiations, similar to non-mobilized BM-MSCs. In a co-culture system, rat M0-type macrophages co-cultured with PB-MSCs have shown higher expression of M2 markers including CD206, Arg-1, IL-10, and CCL-22 than BM-MSCs, indicating that PB-MSCs induced greater M0 polarization to M2. Furthermore, compared with BM-MSCs, PB-MSCs in a co-culture system with lipopolysaccharide-induced M1-type macrophages more efficiently promoted M1 polarization to M2, accompanied by increasing expression of CD206, Arg-1, IL-10, and CCL-22 while decreasing expression of M1 markers including iNOS, TNF-α, IL-1β and IL-6, indicating that PB-MSCs triggered greater M1 polarization to M2. Subsequently, polymerase chain reaction arrays showed higher expressions of both IL1rn and Tnfrsf11b in PB-MSCs versus BM-MSCs. In response to an inflammatory niche, such as TNF-α, PB-MSCs have shown higher expression and release of IL1RA, causing greater M2 polarization of macrophages, and the special effects may be almost entirely abolished through the neutralization antibody of IL1RA. Mechanistic studies determined that PB-MSCs showed higher levels NF-κBp65 and NF-κBp-p65 than BM-MSCs, which could be obviously enhanced by TNF-α. And the increased IL1RA expression by TNF-α in PB-MSCs could be markedly canceled by an NF-κB inhibitor PDTC. Interestingly, mimicking the mobilized PB-MSCs by a combination of G-CSF and AMD3100 in vivo, BM-MSCs were treated with G-CSF and/or AMD3100 in vitro, showing the increased expressions of NF-κBp65 and IL1RA, which could be prominently abolished by PDTC. Therefore, targeting IL1rn, gene modification or drug intervention for MSCs may provide a novel therapeutic strategy for human diseases, especially inflammatory diseases.

Highlights

Inflammation is a common cause of numerous human diseases, such as heart disease, diabetes mellitus, Alzheimer’s disease, hepatitis, acute lung injury, arthritis, and acute pancreatitis
We observed purified PB-Mesenchymal stromal cells (MSCs) and Bone marrow-derived MSCs (BM-MSCs) under an inverted phase-contrast microscope to determine whether cultured peripheral blood-derived MSCs (PB-MSCs) and BM-MSCs exhibited traits of stem cells
Red lipid droplets, red phosphate crystals, and blue nodules were observed in both differentiated PB-MSCs and BM-MSCs

Summary

Introduction

Inflammation is a common cause of numerous human diseases, such as heart disease, diabetes mellitus, Alzheimer’s disease, hepatitis, acute lung injury, arthritis, and acute pancreatitis. IL-1α, IL-1β, and IL-1RA are the most widely studied members of the IL-1 family, and their respective biological activities are pivotal in inflammatory mechanisms for the activation of macrophages (Dinarello, 1996). Both IL-1α and IL-1β interact with the IL-1 receptor 1 (IL-1R1) and recruit the IL-1 receptor accessory protein (IL-1R3, formerly termed IL-1RAcP) to induce a downstream signal via several inflammatory kinases, such as MAPK-P38, ERK, JNK, AKT, and NF-κB, leading to transcription of inflammatory cytokines (Huaux et al, 2015; Sakai et al, 2015; El et al, 2018; Maruyama et al, 2019). Increasing the levels of the members of the IL-1 family (IL-1RA) may be beneficial in inducing the macrophage phenotypic switch toward M2

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