Leptin stimulates interleukin-6 production via janus kinase 2/signal transducer and activator of transcription 3 in rheumatoid synovial fibroblasts.
Leptin stimulates interleukin-6 production via janus kinase 2/signal transducer and activator of transcription 3 in rheumatoid synovial fibroblasts.
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发表时间：
2013-04
影响因子：
3.7
通讯作者：
S. Muraoka;N. Kusunoki;Hiroshi Takahashi;Kazuaki Tsuchiya;S. Kawai
中科院分区：
文献类型：
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作者：
S. Muraoka;N. Kusunoki;Hiroshi Takahashi;Kazuaki Tsuchiya;S. Kawai
本研究的目的是确定瘦素对类风湿性滑膜成纤维细胞（RSFs）产生促炎细胞因子的影响。方法滑膜组织取自类风湿关节炎（RA）患者。逆转录-聚合酶链反应（RT-PCR）检测瘦素受体mRNA。采用实时荧光定量PCR和ELISA试剂盒分别检测培养液中白细胞介素1 β（IL-1 β）、肿瘤坏死因子α（TNF-α）和IL-6的mRNA和蛋白的产生。用小干扰RNA（siRNA）转染RSF，下调瘦素受体的表达。评估Janus激酶2（JAK 2）、磷脂酰肌醇3-激酶（PI 3 K）和丝裂原活化蛋白激酶（MAPK）的抑制剂对IL-6产生的影响。Western blot检测RSF中信号转导子和转录激活子3（STAT 3）的磷酸化水平。结果我们在RSF中检测到瘦素受体mRNA。RSFs中加入瘦素后，IL-1β和IL-6 mRNA的表达呈浓度依赖性增强。瘦素刺激后RSFs分泌IL-6增加。在暴露于靶向瘦素受体（Ob-Rb）的siRNA后，瘦素诱导的RSFs产生IL-6减少。JAK 2抑制剂，而不是PI 3 K和MAPK抑制剂，减少瘦素诱导的IL-6的产生。在瘦素刺激后，观察到RSFs中STAT 3的磷酸化增强。结论瘦素可能是通过激活JAK 2/STAT 3上调RSFs产生IL-6的促炎细胞因子之一。瘦素和JAK/STAT通路可能成为RA治疗的新靶点。
OBJECTIVES The aim of this study was to determine the influence of leptin on the production of proinflammatory cytokines by rheumatoid synovial fibroblasts (RSFs). METHODS Synovial tissue was obtained from patients with rheumatoid arthritis (RA). Leptin receptor mRNAs were detected by reverse transcription-polymerase chain reaction (RT-PCR). Productions of mRNA and protein of interleukin (IL)-1β, tumour necrosis factor-α (TNF-α), and IL-6 in the culture medium were detected by real-time PCR and ELISA kit, respectively. Small interfering RNA (siRNA) was transfected into RSF to down-regulate the expression of leptin receptor. Effects of inhibitors of janus kinase 2 (JAK2), phosphatidylinositol 3-kinase (PI3K), and mitogen-activated protein kinase (MAPK) on IL-6 production were evaluated. Phosphorylation of signal transducer and activator of transcription 3 (STAT3) in RSF were determined by Western blot analysis. RESULTS We detected leptin receptor mRNAs in RSFs. Expression of IL-1β and IL-6 mRNA was enhanced in a concentration-dependent manner by addition of leptin to RSFs. IL-6 secretion by RSFs showed an increase after leptin stimulation. Leptin-induced production of IL-6 by RSFs was decreased after exposure to siRNA targeting leptin receptor (Ob-Rb). A JAK2 inhibitor, but not PI3K and MAPK inhibitors, decreased leptin-induced IL-6 production. Enhanced phosphorylation of STAT3 was observed in RSFs after stimulation by leptin. CONCLUSIONS Leptin may be one of the proinflammatory cytokines that up-regulates IL-6 production in RSFs via activation of JAK2/STAT3. Leptin and JAK/STAT pathway may represent a new alternative therapeutic target in the treatment of RA.