Zou Yaoyao, Zeng Shan, Huang Mingcheng, Qiu Qian, Xiao Youjun, Shi Maohua, Zhan Zhongping, Liang Liuqin, Yang Xiuyan, Xu Hanshi
Department of Rheumatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China.
Br J Pharmacol. 2017 May;174(9):893-908. doi: 10.1111/bph.13762. Epub 2017 Mar 27.
Abnormal glycolytic metabolism contributes to joint inflammation in rheumatoid arthritis (RA). The aims of this study were to investigate the role of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), a bifunctional enzyme that controls the glycolytic rate, in regulating fibroblast-like synoviocyte (FLS)-mediated synovial inflammation and invasiveness in RA.
A specific inhibitor of PFKFB3, PFK15, and siRNA were used to evaluate the role of PFKFB3. Protein expression was measured by Western blotting or immunofluorescence staining. The expression of cytokines was determined by quantitative real-time PCR. Migration and invasion were measured using a Boyden chamber assay. A mouse model of collagen-induced arthritis (CIA) was used to evaluate the in vivo effect of PFK15.
PFKFB3 expression was increased in the synovial tissue and FLSs from RA patients compared with osteoarthritis patients. PFKFB3 inhibition decreased the expression of IL-8, IL-6, CCL-2 and CXCL-10 and the proliferation, migration and invasion of RA FLSs. PFK15 suppressed TNF-α-induced activation of NF-κB and p38, JNK and ERK MAPK signals in RA FLSs. PFK15 treatment also suppressed glucose uptake and lactate secretion. Lactate reversed the inhibitory effect of PFK15 or PFKFB3 siRNA on cytokine expression and migration of RA FLSs. Lactate was also involved in PFKFB3-mediated activation of NF-κB and MAPKs. Intraperitoneal injection of PFK15 in mice with CIA attenuated joint inflammation.
Elevated PFKFB3 expression might contribute to synovial inflammation and aggressive behaviours of RA FLSs, suggesting a novel strategy of targeting PFKFB3 to prevent synovial inflammation and joint destruction in RA.
异常糖酵解代谢促成类风湿关节炎（RA）中的关节炎症。本研究旨在探究6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）（一种控制糖酵解速率的双功能酶）在调节RA中成纤维样滑膜细胞（FLS）介导的滑膜炎症和侵袭性方面的作用。
使用PFKFB3的特异性抑制剂PFK15和小干扰RNA（siRNA）来评估PFKFB3的作用。通过蛋白质印迹法或免疫荧光染色测量蛋白质表达。通过定量实时聚合酶链反应测定细胞因子的表达。使用博伊登小室分析法测量迁移和侵袭。采用胶原诱导性关节炎（CIA）小鼠模型评估PFK15的体内效应。
与骨关节炎患者相比，RA患者滑膜组织和FLS中PFKFB3表达增加。抑制PFKFB3可降低IL-8、IL-6、CCL-2和CXCL-10的表达以及RA FLS的增殖、迁移和侵袭。PFK15抑制RA FLS中肿瘤坏死因子-α（TNF-α）诱导的核因子-κB（NF-κB）以及p38、应激活化蛋白激酶（JNK）和细胞外信号调节激酶（ERK）丝裂原活化蛋白激酶（MAPK）信号通路的激活。PFK15处理还抑制葡萄糖摄取和乳酸分泌。乳酸可逆转PFK15或PFKFB3 siRNA对RA FLS细胞因子表达和迁移的抑制作用。乳酸还参与PFKFB3介导的NF-κB和MAPK激活。对CIA小鼠腹腔注射PFK15可减轻关节炎症。
PFKFB3表达升高可能促成RA FLS的滑膜炎症和侵袭性行为，提示靶向PFKFB3以预防RA滑膜炎症和关节破坏的新策略。
