Acacetin inhibits expression of matrix metalloproteinases via a MAPK‐dependent mechanism in fibroblast‐like synoviocytes

It is well known that rheumatoid arthritis (RA) is an autoimmune joint disease in which fibroblast‐like synoviocytes (FLSs) play a pivotal role. In this study, we investigated the anti‐arthritic properties of acacetin in FLSs. The expression of matrix metalloproteinase (MMP)‐1, MMP‐3 and MMP‐13 were investigated by quantitative RT‐PCR and western blot at gene and protein levels. At the same time, the phosphorylation of mitogen‐activated protein kinases (MAPK) was investigated. The DNA‐binding activity of NF‐κB was investigated by electrophoretic mobility shift assay. We found that acacetin inhibits p38 and JNK phosphorylation and reduces MMP‐1, MMP‐3 and MMP‐13 expression in interleukin‐1β‐induced FLSs. Our results suggest that acacetin has antiarthritic effects in FLSs. Thus, acacetin should be further studied for the treatment of arthritis.     

hTNFRII-Fc融合基因的克隆及在CHO细胞中的表达

构建人肿瘤坏死因子受体II胞外区（TNFRII）和人免疫球蛋白（IgG1）Fc段的融合基因真核表达载体,并在中国仓鼠卵巢细胞（CHO）中表达融合蛋白hTNFRII-Fc。用全合成重叠延伸PCR方法克隆TNFRII胞外区的基因片段,再与（IgG1）Fc段拼接起来形成hTNFRII-Fc融合基因,经HindIII、EcoRI双酶切后插入至pEE14中构建pEE14-TNFRII-Fc真核表达载体,脂质体转染至CHO细胞中进行表达及鉴定。ELISA检测到转染hTNFRII-Fc基因的CHO细胞培养上清中含有hTNFRII-Fc蛋白,SDS-PAGE与Western-blot鉴定证实其为hTNFRII-Fc蛋白,为进一步研究该融合蛋白在CHO细胞稳定表达及应用于类风湿关节炎等自身免疫性疾病临床治疗奠定了一定的研究基础。     

Epstein－Barr病毒核抗原Ⅱ（EBNA2）重组质粒pSG5－EBNA2－Hyg的构建及其在哺乳动物传代细胞中的表达

Ｅｐｔｅｉｎ－Ｂａｒｒ病毒,ＥＢＮＡ２短暂表达,类风湿关节炎     

Pre-analytical and biological variability in circulating interleukin 6 in healthy subjects and patients with rheumatoid arthritis

Abstract

Interleukin (IL)-6, a key player in the inflammatory response, may be a useful biomarker in rheumatoid arthritis (RA). The aim was to determine analytical variability, a reference interval in healthy subjects, and long- and short-term variation in serum and plasma IL-6 in healthy subjects and RA patients. An enzyme-linked immunosorbent assay from R&;D was used for determination of serum and plasma IL-6. The IL-6 concentration did not depend on the type of anticoagulant used or the 3-h time delay between sampling and processing or repeated freeze–thaw cycles. The median plasma and serum IL-6 in 318 healthy subjects were 1.3 pg ml^?1 (range 0.33–26) and 1.4 pg ml^?1 (range 0.25–23), respectively. The median coefficient of variation in plasma IL-6 in 27 healthy subjects during 1 month, and repeated after 6 and 12 months were 27%, 31% and 26%, respectively. No significant long-term changes were observed in serum IL-6 over a 3-year period (14%, p=0.33). Exercise (cycling) increased serum IL-6 in healthy subjects but not in RA patients. In conclusion, circulating IL-6 is stable regarding sample handling and shows little variation over time. Changes in IL-6 concentrations >60% (2 times the biological variation) are likely to reflect changes in disease activity and not only pre-analytical or normal biological variability.     

小剂量来氟米特治疗类风湿关节炎的临床研究

目的：前瞻性观察小剂量来氟米特治疗类风湿关节炎（RA）的临床疗效。方法：32例类风湿关节炎病例随机进入治疗组及对照组。治疗方案治疗组采用小剂量来氟米特（略去负荷剂量,每日剂量10mg维持）,对照组使用柳氮磺胺吡啶治疗,1.5-2.0g/日。观察期6个月,观察指标为主要疗效指标：肿胀、压痛关节数、患者及医师对疾病状况总体评价;次要疗效指标疼痛视觉模拟评分、晨僵时间、美国风湿病学会疗效评价指标（ACR20、50）、健康评价问卷（HAQ）、C反应蛋白。结果：治疗6个月后,主要疗效指标压痛、肿胀关节数改善及医师总体评价,来氟米特均优于柳氮磺胺吡啶（p〈0.05）。在次要疗效指标方面,来氟米特组HAQ评分及C反应蛋白改善优于柳氮磺胺吡啶组（p〈0.05）,两组在关节晨僵改善无明显差异。达到ACR20标准的病例,来氟米特组占56.3%,柳氮磺胺吡啶组占57.1%（p〉0.05）;达到ACR50标准分别为37.5%、35.7%（p〉0.05）。研究中,来氟米特组胃肠道反应轻微,有2例出现血压升高。结论：小剂量来氟米特治疗类风湿关节炎治疗活动性类风湿关节炎,与柳氮磺胺吡啶疗效相当,耐受性好。     

IL-6: TNF-α之后的类风湿关节炎治疗关键靶点

IL-6是一种重要的细胞因子,在类风湿关节炎发病过程中发挥重要作用。本文对已上市的IL-6受体单克隆抗体tocilizumab对类风湿关节炎的临床疗效和安全性进行了总结,并和TNF-α阻断药物进行了对比,证明tocilizumab在药物疗效和副作用方面与TNF-α阻断药物相比各有优劣。另外也对在研的IL-6通路阻断单抗的临床试验结果进行了总结。结合本中心近年的研究和总结,IL-6是继TNF-α之后的另一个重要的类风湿关节炎治疗关键靶点,该类药物的上市为以后类风湿关节炎的个性化治疗提供了更多的选择。     

治疗关节炎症的新靶点——聚蛋白多糖酶Aggrecanase的研究进展

聚蛋白多糖酶（aggrecanase）是新发现的降解软骨组织重要成分聚蛋白多糖（aggrecan）的金属蛋白酶,是治疗关节炎的新靶点.Aggrecanase的发现使得关节炎的治疗有望取得新的突破,为能从根本上治疗和预防关节炎提供了基础.综述了aggrecanase从克隆到对其性质的研究,阐明以该酶为靶点寻找抑制剂是治疗与预防关节炎的一条可能的途径.     

Classifying Integrated Signature Molecules in Macrophages of Rheumatoid Arthritis,Osteoarthritis, and Periodontal Disease: An Omics-Based Study

Rheumatoid arthritis (RA), osteoarthritis (OA), and periodontal disease (PD) are chronic inflammatory diseases that are globally prevalent, and pose a public health concern. The search for a potential mechanism linking PD to RA and OA continues, as it could play a significant role in disease prevention and treatment. Recent studies have linked RA, OA, and PD to Porphyromonas gingivalis (PG), a periodontal bacterium, through a similar dysregulation in an inflammatory mechanism. This study aimed to identify potential gene signatures that could assist in early diagnosis as well as gain insight into the molecular mechanisms of these diseases. The expression data sets with the series IDs {"type":"entrez-geo","attrs":{"text":"GSE97779","term_id":"97779"}}GSE97779, {"type":"entrez-geo","attrs":{"text":"GSE123492","term_id":"123492"}}GSE123492, and {"type":"entrez-geo","attrs":{"text":"GSE24897","term_id":"24897"}}GSE24897 for macrophages of RA, OA synovium, and PG stimulated macrophages (PG-SM), respectively, were retrieved and screened for differentially expressed genes (DEGs). The 72 common DEGs among RA, OA, and PG-SM were further subjected to gene–gene correlation analysis. A GeneMANIA interaction network of the 47 highly correlated DEGs comprises 53 nodes and 271 edges. Network centrality analysis identified 15 hub genes, 6 of which are DEGs (API5, ATE1, CCNG1, EHD1, RIN2, and STK39). Additionally, two significantly up-regulated non-hub genes (IER3 and RGS16) showed interactions with hub genes. Functional enrichment analysis of the genes showed that “apoptotic regulation” and “inflammasomes” were among the major pathways. These eight genes can serve as important signatures/targets, and provide new insights into the molecular mechanism of PG-induced RA, OA, and PD.     

高脂食物对动脉硬化合并类风湿关节炎小鼠肠道菌群的影响

目的 分析高脂食物对动脉硬化合并类风湿关节炎小鼠肠道微生物的影响,了解动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。方法 8周龄ApoE-/-小鼠饲喂高脂食物和普通食物至17周龄来诱发动脉硬化症状,再通过给17周龄ApoE-/-小鼠腹腔注射抗6-磷酸葡萄糖异构酶（glucose-6-phosphate isomerase,GPI）抗体呈阳性的K/BxN血清,从而诱导其产生类风湿关节炎症状。通过Illumina HiSeq平台对各组小鼠粪便进行16S rDNA V4区测序,分析动脉硬化合并类风湿关节炎小鼠肠道微生物的变化。结果 ApoE-/-小鼠饲喂高脂食物后,其血清低密度脂蛋白胆固醇（LDL-C）浓度和血清总胆固醇（TC）浓度均显著升高,主动脉内膜斑块面积比喂普通食物的ApoE-/-小鼠显著增加,表明ApoE-/-小鼠饲喂高脂食物后引起更显著的动脉硬化症状。再通过腹腔注射抗GPI抗体呈阳性的K/BxN血清,各组ApoE-/-小鼠均出现关节肿胀,饲喂高脂食物的ApoE-/-小鼠其踝关节宽度和临床评分（clinical score）低于饲喂普通食物组小鼠。OTU数、Shannon指数和Simpson指数显示高脂食物和K/BxN血清处理组ApoE-/-小鼠肠道菌群多样性降低,Firmicutes/Bacteroidetes值升高,t-test分析显示在属水平上,Prevotellaceae_UCG-001显著降低,Ruminiclostridium_6显著升高。t-test分析和Firmicutes/Bacteroidetes比值显示ApoE-/-小鼠肠道菌群结构紊乱。结论 高脂食物使ApoE-/-小鼠的肠道菌群组成和结构发生改变,导致ApoE-/-小鼠的动脉硬化症状加重,类风湿关节炎症状减轻。提示肠道微生物组成和结构的改变,可能与动脉硬化合并类风湿关节炎发病机制相关。     

Synthetic TGF-β Signaling Agonist-Treated Dendritic Cells Induce Tolerogenicity and Antirheumatic Effects

The newly synthesized compound TGF-β signaling agonist (T74) is a small molecule associated with the TGF-β receptor signaling pathway. Tolerogenic dendritic cells (tDCs) have been used to examine immunosuppressive and anti-inflammatory effects in multiple autoimmune disease models. The aim of this study was to investigate whether treatment of DCs with T74 has an antirheumatic effect in a mouse model of collagen-induced arthritis (CIA). Bone marrow-derived cells were obtained from DBA/1J mice and differentiated into DCs. T74-treated DCs (T74-DCs) were generated by treating bone marrow-derived DCs with LPS, type II collagen, and T74. T74-DCs expressed lower levels of surface molecules and inflammatory cytokines associated with antigen presentation and T cell stimulation. The ability of T74-DCs to differentiate effector T cells was lower than that of T74-untreated DCs (NT-DCs), but T74-DCs increased the regulatory T (Treg) cell differentiation in vitro. DBA/1J mice received two subcutaneous (s.c.) injections of type II collagen to establish CIA. Mice then received two s.c. injections of T74-DCs or NT-DCs. Joint inflammation was ameliorated in the paws of T74-DC-treated mice. Additionally, Treg populations in T74-DC-treated mice were higher than in NT-DC-treated or PBS-treated CIA mice. Taken together, these results demonstrate that T74 induces tolerance in DCs, and that T74-mediated DCs exert antirheumatic effects via induction of Tregs.