Objective To investigate the effect of Miao medicine Jinwu Jiangu decoction containing serum freeze-dried powder on levels of IL-17,ACT1 and TRAF6 in human rheumatoid arthritis fibroblast like synoviocytes (RA-HFLS). Methods Rabbits were randomly divided into blank control group (recieving normal saline of the same volume),Jinwu Jiangu decoction high-dose,medium-dose and low-dose group (intragastrically administrated with Jinwu Jiangu decoction at doses of 14.4,4.8 and 2.4 g·kg-1,respectively),tripterygium glycosides group and prednisone group (treated with human equivalent dosage).RA-HFLS primary cell model was established in the experiment.ELISA method was used to detect effect of lyophilized powder on IL-17 secretion.Expression of ACT1,TRAF6 mRNA was detected by RT-PCR. Results Compared with the blank control gorup,IL-17 in the supernatant of each medication administration group was significantly decreased (all P<0.01),and it was decreased most significantly in Jinwu Jiangu decoction high-dose and medium-dose group.IL-17 was down-regulated more significantly in high-dose group than that in tripterygium glycosides group (P<0.01).Compared with the blank control group,TRAF6 and ACT1 mRNA expression level of each medication administration group were significantly decreased (all P<0.01),and in the high-dose group that were decreased most significantly,but not significantly different as compared with tripterygium glycosides group and prednisone group (P>0.05). Conclusion Freeze-dried powder of Jinwu Jiangu decoction can decrease the secretion of IL-17 and down-regulate expression of ACT1,TRAF6 with RA-HFLS.
表1
金乌健骨方含药血清对RA-HFLS培养上清液中IL-17分泌的影响
Tab.1
Effects of Jinwu Jiangu decoction containing serum on IL-17 secretion in the supernatant of cultured RA-HFLS x¯±s,n=5
组别
剂量/(g·kg-1)
IL-17/(ng·L-1)
空白对照组
-
17.50±0.67
雷公藤多苷组
0.36
3.15±0.53*1
金乌健骨方小剂量组
2.4
9.71±0.51*1
金乌健骨方中剂量组
4.8
2.64±0.30*1*2
金乌健骨方大剂量组
14.4
1.97±0.28*1*2
Compared with blank control group,*1P<0.01;compared with tripterygium glycosides group,*2P<0.01
与空白对照组比较,*1P<0.01;与雷公藤多苷组比较,*2P<0.01
表1
金乌健骨方含药血清对RA-HFLS培养上清液中IL-17分泌的影响
Tab.1
Effects of Jinwu Jiangu decoction containing serum on IL-17 secretion in the supernatant of cultured RA-HFLS x¯±s,n=5
Fig.1 mRNA expression of TRAF6 in six groups of RA-HFLS K.blank control group;1.low-dose Jinwu Jiangu decoction group;2.medium-dose Jinwu Jiangu decoction group; 3.high-dose Jinwu Jiangu decoction group;4. tripterygium glycosides group;5.prednisone group
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Interleukin-17 (IL-17) is critically involved in the pathogenesis of various inflammatory disorders. IL-17 receptor (IL-17R)-proximal signaling complex (IL-17R-Act1-TRAF6) is essential for IL-17-mediated NF-κB activation, while IL-17-mediated mRNA stability is TRAF6 independent. Recently, inducible IκB kinase (IKKi) has been shown to phosphorylate Act1 on Ser 311 to mediate IL-17-induced mRNA stability. Here we show that TANK binding kinase 1 (TBK1), the other IKK-related kinase, directly phosphorylated Act1 on three other Ser sites to suppress IL-17R-mediated NF-κB activation. IL-17 stimulation activated TBK1 and induced its association with Act1. IKKi also phosphorylated Act1 on the three serine sites and played a redundant role with TBK1 in suppressing IL-17-induced NF-κB activation. Act1 phosphorylation on the three sites inhibited its association with TRAF6 and consequently NF-κB activation in IL-17R signaling. Interestingly, TRAF6, but not TRAF3, which is the upstream adaptor of the IKK-related kinases in antiviral signaling, was critical for IL-17-induced Act1 phosphorylation. TRAF6 was essential for IL-17-induced TBK1 activation, its association with Act1, and consequent Act1 phosphorylation. Our findings define a new role for the IKK-related kinases in suppressing IL-17-mediated NF-κB activation through TRAF6-dependent Act1 phosphorylation.
SAKUMAM, HATSUSHIKAK, KOYAMAK, et al.TGF-β type I receptor kinase inhibitor down-regulates rheumatoid synoviocytes and prevents the arthritis induced by type II collagen antibody[J]. Int Immunol, 2007, 19(2): 117-126.
Rheumatoid arthritis (RA) is characterized by hypertrophic synovial tissues comprising excessively proliferating synovial fibroblasts and infiltrating inflammatory cells. Transforming growth factor-β (TGF-β) is a multifunctional cytokine that regulates cell growth, inflammation and angiogenesis by acting on various cell types. In RA synovial tissues, TGF-β is expressed at high levels. However, the precise role of TGF-β in RA remains unclear. We herein demonstrated a causal link between the TGF-β-induced RA synovial cell proliferation and induction of platelet-derived growth factor (PDGF)-AA. In addition, TGF-β induced IL-6 and vascular endothelial growth factor (VEGF) production by RA synovial fibroblasts associated with nuclear factor-kappa B activation. These effects of TGF-β on RA synovial fibroblasts were suppressed by TGF-β type I receptor kinase inhibitor HTS466284. Furthermore, HTS466284 significantly prevented anti-collagen type II antibody-induced arthritis in mice according to the clinical manifestations, histology, tumor necrosis factor-α, PDGF and VEGF expression and 5-bromo-2′-deoxyuridine incorporation. These in vitro and in vivo results suggest that TGF-β plays a role in the development of synovial hyperplasia consisting of synovial cell proliferation, inflammation and angiogenesis. The blockade of TGF-β signaling may thus become an additional strategy for the treatment of RA.
NAKAES, NAMBUA, SUDOK, et al.Suppression of immune induction of collagen-induced arthritis in IL-17-deficient mice[J].Immunol,2003,171(11):6173-6177.
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LUBBERTSE, KOENDERS M I. OPPERS W B, et al.Treatment with a neutralizing anti-murine interleukin-17 antibody after the onset of collagen-induced arthritis reduces joint inflammation, cartilage destruction, and bone erosion[J].Arthritis Rheum,2004,50(2):650-659.
Abstract Top of page Abstract MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES Objective Interleukin-17 (IL-17) is a proinflammatory cytokine that is expressed in the synovium of rheumatoid arthritis (RA) patients. This T cell cytokine is implicated in the initiation phase of arthritis. However, the role of IL-17 during the effector phase of arthritis has still not been identified; this was the objective of the present study. Methods Mice with collagen-induced arthritis (CIA) were treated with polyclonal rabbit anti-murine IL-17 (anti–IL-17) antibody–positive serum or normal rabbit serum after the first signs of arthritis. In addition, during a later stage of CIA mice were selected and treated with anti–IL-17 antibody or control serum. Arthritis was monitored visually, and joint pathology was examined radiologically and histologically. Systemic IL-6 levels were measured by enzyme-linked immunosorbent assay, and local synovial IL-1 and receptor activator of NF-κB ligand (RANKL) expression was analyzed using specific immunohistochemistry. Results Treatment with a neutralizing anti–IL-17 antibody after the onset of CIA significantly reduced the severity of CIA. Radiographic analysis revealed marked suppression of joint damage in the knee and ankle joints. Histologic analysis confirmed the suppression of joint inflammation and showed prevention of cartilage and bone destruction after anti–IL-17 antibody therapy. Systemic IL-6 levels were significantly reduced after anti–IL-17 antibody treatment. Moreover, fewer IL-1β–positive and RANKL-positive cells were detected in the synovium after treatment with neutralizing IL-17. Interestingly, initiation of anti–IL-17 antibody therapy during a later stage of CIA, using mice with higher clinical arthritis scores, still significantly slowed the progression of the disease. Conclusion IL-17 plays a role in early stages of arthritis, but also later during disease progression. Systemic IL-6 was reduced and fewer synovial IL-1–positive and RANKL-positive cells were detected after neutralizing endogenous IL-17 treatment, suggesting both IL-1–dependent and IL-1–independent mechanisms of action. Our data strongly indicate that IL-17 neutralization could provide an additional therapeutic strategy for RA, particularly in situations in which elevated IL-17 may attenuate the response to anti–tumor necrosis factor/anti–IL-1 therapy.
LUBBERTSE, Joosten LA, VAN DE LOO F A. et al. Overexpression of IL-17 in the knee joint of collagen type II immunized mice promotes collagen arthritis and aggravates joint destruction[J].lnflamm Res,2002,51(2):102-104.
ErbB-4 is a receptor tyrosine kinase that is activated by the binding of specific growth factors to its ectodomain. In addition to the initiation of signal transduction pathways that direct cell responses, such as proliferation or differentiation, this receptor is subject to ligand-dependent trafficking events. The signal transduction events are controlled by ligand-dependent activation of the receptor tyrosine kinase activity, which results in receptor autophosphorylation and the tyrosine phosphorylation of other cellular proteins. The trafficking events include migration into and out of membrane microdomains, entry into internalization pathways and endocytosis, plus proteolytic fragmentation.
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KEHLENA, THIELEK, RIEMANND, et al.Expression,modulation and signalling of IL-17 receptor in fibroblast-like synoviocytes of patients with rheumatoid arthritis[J].Clin Exp Immunol,2002,127(3):539-546.
SUMMARY Interleukin-17 (IL-17) has been characterized as a proinflammatory cytokine produced by CD4CD45ROmemory T cells. Overproduction of IL-17 was detected in the synovium of patients with rheumatoid arthritis (RA) compared to patients with osteoarthritis. In contrast to the restricted expression of IL-17, the IL-17 receptor (IL-17R/CDw217) is expressed ubiquitously. Using a real-time RT-PCR assay, we detected similar absolute levels of IL-17R mRNA expression in fibroblast-like synoviocytes (SFC) from patients with RA (mean 9 pg/μg total RNA; ranged from 0·1 pg to 96 pg IL-17R mRNA/μg total RNA) compared to synoviocytes of non-RA patients. Analysis of the IL-17R surface expression confirmed the results obtained for IL-17R mRNA expression. Exposure of SFC to IL-17 led to a mRNA induction of CXC chemokines IL-8, GRO-α and GRO-β. An anti-IL-17 antibody blocked these effects of IL-17. The MAPK p38 appears necessary for the regulation of IL-8, GRO-α and GRO-β expression as shown by inhibition with SB203580. The inhibitors genistein (tyrosine kinase inhibitor) and calphostin C (inhibitor of protein kinase C) reduced significantly the IL-17-stimulated mRNA expression of IL-8, GRO-α and GRO-β in SFC, whereas PD98059 (inhibitor of MEK-1/2) was without effect. Pharmacological drugs used in therapy of RA, such as cyclosporin and methotrexate, induced a fourfold increase of IL-17R mRNA expression and augmented the IL-17-stimulated IL-8 expression. Our results support the hypothesis that IL-17/IL-17R may play a significant role in the pathogenesis of RA contributing to an unbalanced production of cytokines as well as participating in connective tissue remodelling.
ANNAS, MARIANS.IL-17-expressing cells as a potential therapeutictarget for treatment of immunological disorders[J]. Pharmacol Rep,2011,63(1): 30-44.
IL-17 is a multifunctional cytokine produced by activated CD4+ and CD8+ lymphocytes as well as stimulated unconventional T纬未 and natural killer T cells. IL-17 induces expression of chemokines, proinflammatory cytokines and metalloproteinases, thereby stimulating the inflammation and chemotaxis of neutrophils. Elevation of proinflammatory cytokines is associated with asthma and autoimmune disorders, such as multiple sclerosis, rheumatoid arthritis and psoriasis. Although the role of IL-17 in these disorders is not always easy to define, extensive research has demonstrated an aggravating influence of IL-17 in some animal models. Thus, the development of therapeutics to reduce IL-17 levels is a promising strategy for ameliorating inflammatory diseases. This review briefly summarizes recent knowledge about stimulants and intracellular signaling pathways that induce development and maturation of IL-17-expressing cells. Its positive and negative roles on disease progression and its importance in vaccineinduced memory are also discussed. Finally, recent literature describing potential therapeutic approaches for targeting IL-17 is presented.
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SEHWANDNERR, YAMAFUEHIK, CAOZ.Requirernent of tumor neerosis factor receptor-associated factor(TRAF)6 in interleukin-17 signal transduetion[J].Exp Med,2000,191(7):1233-1240.
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FANGFANGQ, HANAHAOG, SHUZ, et al.TRAF6-dependent Act1 phosphorylation by the IκB kinase-related kinases suppresses interleukin-17-induced NF-κB activation[J].Mol Cell Biol,2012,32(19):3925-3937.
Interleukin-17 (IL-17) is critically involved in the pathogenesis of various inflammatory disorders. IL-17 receptor (IL-17R)-proximal signaling complex (IL-17R-Act1-TRAF6) is essential for IL-17-mediated NF-κB activation, while IL-17-mediated mRNA stability is TRAF6 independent. Recently, inducible IκB kinase (IKKi) has been shown to phosphorylate Act1 on Ser 311 to mediate IL-17-induced mRNA stability. Here we show that TANK binding kinase 1 (TBK1), the other IKK-related kinase, directly phosphorylated Act1 on three other Ser sites to suppress IL-17R-mediated NF-κB activation. IL-17 stimulation activated TBK1 and induced its association with Act1. IKKi also phosphorylated Act1 on the three serine sites and played a redundant role with TBK1 in suppressing IL-17-induced NF-κB activation. Act1 phosphorylation on the three sites inhibited its association with TRAF6 and consequently NF-κB activation in IL-17R signaling. Interestingly, TRAF6, but not TRAF3, which is the upstream adaptor of the IKK-related kinases in antiviral signaling, was critical for IL-17-induced Act1 phosphorylation. TRAF6 was essential for IL-17-induced TBK1 activation, its association with Act1, and consequent Act1 phosphorylation. Our findings define a new role for the IKK-related kinases in suppressing IL-17-mediated NF-κB activation through TRAF6-dependent Act1 phosphorylation.
Treatment with a neutralizing anti-murine interleukin-17 antibody after the onset of collagen-induced arthritis reduces joint inflammation, cartilage destruction, and bone erosion
2004
VAN DE LOO F A. et al. Overexpression of IL-17 in the knee joint of collagen type II immunized mice promotes collagen arthritis and aggravates joint destruction
, 宁乔怡, 李东晓, 姚血明, 侯雷, 梁江, 黄颖, 周静
