Sphingosine-1-phosphate (S1P) enhances glomerular
endothelial cells activation mediated by anti-myeloperoxidase
a, b, c, d
, Min Chen
a, b, c, d
*, Ming-Hui Zhao
a, b, c, d, e
Renal Division, Department of Medicine, Peking University First Hospital, Beijing, China
Institute of Nephrology, Peking University, Beijing, China
Key Laboratory of Renal Disease, Ministry of Health of China, Beijing, China
Key Laboratory of Chronic Kidney Disease Prevention and Treatment (Peking University), Ministry of Education, Beijing, China
Peking-Tsinghua Center for Life Sciences, Beijing, China
Received: June 23, 2017; Accepted: October 16, 2017
Cumulating evidences suggested an important role of sphingosine-1-phosphate (S1P) and its receptors in regulating endothelial barrier integ-
rity. Our previous study revealed that the circulating S1P levels and renal expression of S1PRs correlated with disease activity and renal damage
in patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). This study investigated the role of S1P and its recep-
tors in myeloperoxidase (MPO)-ANCA-positive IgG-mediated glomerular endothelial cell (GEnC) activation. The effect of S1P on morphological
alteration of GEnCs in the presence of MPO-ANCA-positive IgG was observed. Permeability assay was performed to determine endothelial
monolayer activation in quantity. Both membrane-bound and soluble ICAM-1 and VCAM-1 levels were measured. Furthermore, antagonists
and/or agonists of various S1PRs were employed to determine the role of different S1PRs. S1P enhanced MPO-ANCA-positive IgG-induced dis-
ruption of tight junction and disorganization of cytoskeleton in GEnCs. S1P induced further increase in monolayer permeability of GEnC mono-
layers in the presence of MPO-ANCA-positive IgG. S1P enhanced MPO-ANCA-positive IgG-induced membrane-bound and soluble ICAM-1/
VCAM-1 up-regulation of GEnCs. Soluble ICAM-1 levels in the supernatants of GEnCs stimulated by S1P and MPO-ANCA-positive IgG increased
upon pre-incubation of S1PR1 antagonist, while pre-incubation of GEnCs with the S1PR1 agonist down-regulated sICAM-1 level. Blocking
S1PR2-4 reduced sICAM-1 levels in the supernatants of GEnCs stimulated by S1P and MPO-ANCA-positive IgG. Pre-incubation with S1PR5
agonist could increase sICAM-1 level in the supernatants of GEnC stimulated by S1P and MPO-ANCA-positive IgG. S1P can enhance
MPO-ANCA-positive IgG-mediated GEnC activation through S1PR2-5.
ANCA-AAV is a group of systemic autoimmune diseases, including
microscopic polyangiitis (MPA), granulomatosis with polyangiitis
(GPA) and eosinophilic granulomatosis with polyangiitis (EGPA) .
AAV is characterized by necrotizing inﬂammation of the small blood
vessels, which involves GEnC injury in particular. ANCAs against MPO
and proteinase 3 (PR3) are the serological hallmarks of AAV [2, 3].
What is noteworthy is that Chinese AAV patients, as demonstrated in
our previous studies, are predominantly MPO-ANCA-positive [4, 5].
Moreover, cumulating evidences reveal that MPO-ANCAs can induce
direct GEnC activation and injury in AAV. Nagao et al. [6, 7] reported
that MPO-ANCA could induce up-regulation of adhesion molecules in
GEnCs both in vivo and in vitro.
S1P is a bioactive sphingolipid and key regulator in vascular
inﬂammation. S1P is the ligand for ﬁve G-protein-coupled receptors
(GPCRs) named S1PR1-5 [8, 9]. S1P and its receptors are involved in
the pathogenesis of a variety of vascular inﬂammatory conditions
including sepsis, atherosclerosis and ischaemia–reperfusion injury
[10–14]. In our previous study, it was found that the levels of circulat-
ing S1P were elevated and renal expression of S1PR2-5 was up-regu-
lated in AAV patients of active stage, and that the circulating S1P
levels and the renal expression of S1PR were associated with renal
involvement and disease activity of AAV . Moreover, we found
*Correspondence to: Min CHEN
ª 2017 The Authors.
Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.
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J. Cell. Mol. Med. Vol 22, No 3, 2018 pp. 1769-1777