ABSTRACT
Background and Objectives@#The immunomodulatory potential of mesenchymal stem cells (MSCs) can be regulated by a variety of molecules, especially cytokines. The inflammatory cytokine, TNF-like ligand 1A (TL1A), has been reported as an inflammation stimulator in-multiple autoimmune diseases. Here, we studied the effects of TL1A/TNF-receptor 2 (TNFR2) pathway on the therapeutic potency of bone marrow-derived MSCs (BMSCs). @*Methods@#and Results: BMSCs, fibroblast-like synoviocytes (FLSs), and H9 and jurkat human T lymphocytes were used in this study. BMSCs paracrine activities, differentiation, proliferation, and migration were investigated after stimulation with TL1A, and intervened with anti-TNFR2. Additionally, the effects of TL1A on BMSCs therapeutic potency were evaluated by treating RA-FLSs, and H9 and jurkat T cells with TL1A-stimulated BMSCs conditioned medium (CM). Indian hedgehog (IHH) involvement was determined by gene silencing and treatment by recombinant IHH (rIHH). TL1A induced BMSCs stemness-related genes, COX-2, IL-6, IDO, TGF-β and HGF through TNFR2. Also, TL1A corrected biased differentiation and increased proliferation, and migration through TNFR2. Meanwhile, CM of TL1A-stimulated BMSCs decreased the inflammatory markers of RA-FLSs and T cells. Moreover, TL1A-stimulated BMSCs experienced IHH up-regulation coupled with NF-κB and STAT3 signaling up-regulation, while p53 and oxidative stress were down-regulated. Furthermore, treatment of BMSCs by rIHH increased their anti-inflammatory effects.More importantly, knockdown of IHH decreased the ability of TL1A-stimulated BMSCs to alleviating the inflammation in RA-FLSs and T cells. @*Conclusions@#This study reports the effects of TL1A/TNFR2 pathway on the biological behaviors and therapeutic potency of BMSCs through IHH. These findings could introduce novel procedures to increase the stemness of MSCs in cellular therapy.
ABSTRACT
Objective To investigate the effects of interleukin-34 (IL-34) on prostaglandin E2 (PGE2)/cyclo-oxygenase-2 (COX-2) expression on fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA). Methods FLS was isolated from 6 RA patients and stimulated with IL-34 (50 ng/ml), IL-34 receptor antagonist (25 ng/ml) and IL-34 (50 ng/ml), inhibitors of signaling pathway (10 μmol/L) and IL-34 (50 ng/ml) in vitro respectively. The expression of COX-2 mRNA was detected by reverse transcription polymerase chain reac-tion (RT-PCR). The level of PGE2 in the supernatant of RA FLS culture was measured by Enzyme linked immunosorbent assay (ELISA). Statistical analysis between groups were performed by t test. Results Com-pared to unstimulated FLS, COX-2 and PGE2 expression was increased dramatically on IL-34-stimulated FLS, most evidently in 48 hours [(139±24) pg/ml vs (201±8) pg/ml, t=-6.177, P<0.01]; Moreover, the level of PGE2 was decreased when anti-IL-34 antibody was added to the IL-34-stimulated RA FLS at 24 hours, 48 hours, 72 hours [(250 ±58) pg/ml vs (100 ±28) pg/ml, t=5.742, P<0.01; (375 ±24) pg/ml vs (97 ±23) pg/ml, t=20.564, P<0.001; (357 ±21) pg/ml vs (94 ±18) pg/ml, t=22.353, P<0.01]; In the presence of SB203580 and IKK-16, PGE2 level produced by IL-34-stimulated FLS was obviously decreased [(279 ±37) pg/ml vs (63 ±17) pg/ml, t=12.806, P<0.01;(279±37) pg/ml vs (77±16) pg/ml, t=6.177, P<0.01]. Conclusion Binding of IL-34 with its receptor may promote the secretion of PGE2 via NF-κB and P38 MAPK signaling pathway in RA FLS, suggesting that it might be involved in the pathogenesis of RA.
ABSTRACT
Objective To preliminarily investigate the levels of interleukin (IL)-1 family and IL-34 in serum of patients with ankylosing spondylitis (AS) and their roles.Methods Serum IL-1 family levels were detected from 6 AS patients and 4 healthy controls by using protein-chip technique.Enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of serum IL-34 from 65 AS patients and 85 healthy controls and the relationships of serum IL-34 levels and clinical or laboratory features were analyzed.T test and Spearman correlation were used for statistical analysis.Results IL-1Ra [(3302±1352) pg/ml vs (10778±2764) pg/ml]and IL-36Ra [(1363±194) pg/ml vs (3875±996) pg/ml] levels were significantly down-regulated in AS patients compared with that of healthy controls (t=5.363 and 4.289 respectively,both P＜0.05).The levels of IL-1α,IL-18,IL-36α and IL-37 were increased more remarkable in AS patients than in healthy controls (t=-2.532,-5.400,-5.023 and-5.783 respectively,both P＜0.05).Moreover,serum IL-34 levels were elevated more significantly in AS patients than in healthy controls [(169±153) pg/ml vs (54±31) pg/ml,t=6.722,P＜0.01] and were positively correlated with the levels of CRP and ESR.Serum IL-34 levels were markedly up-regulated in human leukocyte antigen (HLA)-B27 positive patients than in HLA-B27 negative patients(P＜0.05).Conclusion Part of IL-1 family and IL-34 may be involved in inflammatory or immunological process of AS.