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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.
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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.
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Objective To investigate the effects of interleukin (IL)-34 on the proliferation and chemokines expression of fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA).Methods RA FLS were isolated and cultured in the presence or absence of IL-34.FLS proliferation was determined by methyl thiazlyl tetrazoliu method (MTT) method.The levels of chemokines from RA FLS supernatant were detected by protein chip AAH-CYT-G1000 assay.The IL-34 receptor (IL-34R) expression on RA FLS was analyzed by flow cytometry (FCM).Statistical analysis between groups was performed by t test.Results Compared to the control group, the proliferation of IL-34-stimulated RA FLS was obviously increased, and up to the maximum at 72 hours.In addition, the levels of chemokines [epithelial neutrohil-activating peptide 78 (ENA-78), IL-8, growth-related oncogene (GRO), macrophage chemoattractant protein-1 (MCP-1)] on RA FLS supernatant in IL-34 stimulated group were significantly elevated than those in the control group [ENA-78: (397.1±8.2) pg/ml, (54.0±2.9) pg/ml (t=127.61, P＜0.01);IL-8:(2 017±36) pg/ml, (778±102) pg/ml (t=36.67, P＜0.01);GRO: (4 935±160) pg/ml, (2 746±188) pg/ml (t=43.60, P＜0.01);MCP-1: (46 798±1 293) pg/ml, (27 813±2 329) pg/ml (t=22.43, P＜0.01)].IL-34R was highly expressed on the RA FLS.Conclusion IL-34 promotes RA FLS proliferation and chemokines expression, which may be one of the important mechanisms involved in the pathogenesis of RA.
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AIM: To apply molecular systematic techniques to revealing the genetic diversity of medicinal plant of Dendrobium candidum and its related species in Dendrobium.METHODS: The internal transcribed spacer(ITS) as well as 5.8S rDNA sequences of 40 samples of 25 species of Dendrobium were carried out on 25 dendrobi-um samples and amplified using PCR method and sequenced.Mega 3.1 was used to analyze the genetic diversity within the genus.Phylogenetic relationships among species in Dendrobium were estimated by maximum parsimony and neighbor-joining methods to construct similar ITS trees.RESULTS: The phylogenetic analysis indicated that Dendrobium candidum had a markedly difference in the interspecies and possessed the characteristic sequence site.CONCLUSION: ITS sequences as a good molecular marker for authentication between Dendrobium and outgroup is feasibility.