Wnt signaling in rheumatoid synoviocyte activation

Rheumatoid arthritis (RA) is a joint-specific disease with complex pathogenesis. It is characterized by synovial inflammation, cartilage loss, and joint destruction. The reasons why joint damage recurs when therapy is discontinued are not clearly understood. Several lines of evidence suggest that cartilage damage is promoted by the transformed and invasive fibroblast-like synoviocytes (FLS) of the rheumatoid joint. It has been demonstrated in several systems that aberrant wnt-mediated signaling causes blockade of cartilage differentiation and malformation of joints. In this review, we have discussed the importance of wnt–frizzled-mediated signaling in the autonomous activation of FLS in patients with RA. Anti-wnt/anti-frizzled antibodies, frizzled receptor antagonists, or small molecule inhibitors of wnt–frizzled signaling might be useful for therapeutic interventions in RA. Received: May 15, 2001/Accepted: September 28, 2001     

芍甘附子汤对AA大鼠免疫调节及关节滑膜超微结构形态的影响

目的　观察芍甘附子汤对佐剂性关节炎(AA)大鼠免疫调节作用及关节滑膜超微结构形态的影响 ,初步探讨其治疗RA的机理。方法　将 40只SD系大鼠随机分为空白对照组、模型组、芍甘附子汤治疗组及痹颗粒治疗组。除空白对照组外 ,其余三组均复制成完全弗氏佐剂性关节炎模型 ,于造模 1 8天后灌胃给药 ,连续 35天 ,采血测定RBC -C3bR花环率和RBC -IC花环率 :处死动物 ,取胸腺、脾脏称重 ;取右膝关节组织观察病理改变。结果　芍甘附子汤治疗组血中RBC -C3bR花环率升高 ,RBC -IC花环率降低 ,关节滑膜病理损害明显改善 ,与模型组比较差异有显著性意义 ,但对免疫器官无明显影响。结论　芍甘附子汤能够提高AA大鼠RBC免疫活性 ,增强RBC清除免疫复合物的能力 ,减轻其对关节滑膜的侵害     

补肾活血中药含药血清对滑膜细胞分泌TNF-α、IL-1β水平的影响

目的:探讨补肾活血中药是否可以抑制兔滑膜细胞分泌TNF-α、IL-1β等炎性因子。方法:制作补肾活血方含药血清,体外分离并培养兔滑膜细胞,并鉴定、传代。实验分含空白血清对照组、含10%西乐葆血清组、含5%、10%、20%中药血清组等5组。以西乐葆含药血清作为对照,将不同浓度的中西药血清加入培养传代的第3代滑膜细胞,继续培养。观察药物对滑膜细胞分泌TNF-α、IL-1β水平的影响。结果:补肾活血方含药血清具有和西乐葆含药血清类似的抑制滑膜细胞分泌TNF-α、IL-1β的作用,其抑制TNF-α分泌的作用随药物浓度的增加作用增强。结论:补肾活血中药可抑制滑膜细胞分泌TNF-α、IL-1β等炎性因子。     

The synovial production of collagenase and chondrocyte activating factors in response to cobalt

Addition of CoCl2 solutions to the culture media of confluent monolayers of lapine or human synoviocytes stimulated their production of the neutral proteinases collagenase, gelatinase, and caseinase. With lapine cells, maximum stimulation occurred at 10(-7) M CoCl2, while human cells required 10(-4)-10(-5) M CoCl2 to achieve a maximum stimulation. Production of prostaglandin E2 by lapine cells was enhanced some 30-40% by concentrations of CoCl2 that maximally stimulated synthesis of the neutral proteinases, whereas all concentrations of CoCl2 slightly depressed the production of prostaglandin E2 by human cells. Lapine synovial cells that had been stimulated by CoCl2 also produced a substance, or substances, that provoked the synthesis of collagenase, gelatinase, caseinase, and prostaglandin E2 by monolayers of articular chondrocytes. Chondrocytes themselves, however, resisted activation by CoCl2. These findings may be relevant to the aseptic loosening of joint prostheses.     

白细胞介素-18在骨关节炎滑膜细胞中的表达及意义

目的 了解白细胞介素(IL)-18在骨关节炎(OA)患者和非OA患者滑膜细胞中表达的差异.方法 取OA患者(22例)与非OA患者(8例)关节滑膜组织,分别原代培养滑膜细胞,采用酶联免疫吸附测定法(ELISA)定量检测2组滑膜细胞培养上清液中IL-18蛋白的表达水平,采用免疫细胞化学方法 定性观察IL-18在2组滑膜细胞问表达的差异,统计学处理采用t检验.结果 ELISA检测OA患者细胞培养上清液中IL-18的含量为(63.9±21.4)pg/ml, 非OA患者细胞培养上清液中IL-18的含量为(17.7±1.5)pg/ml,2组比较差异有统计学意义(t=10.044,P<0.01);免疫细胞化学方法 显示在OA患者滑膜细胞中IL-18蛋白的表达呈阳性.胞质为棕黄色深染,而在非OA患者滑膜细胞中胞质淡染.结论 IL-18在OA关节滑膜细胞中表达上调, 提示IL-18可能直接参与了OA的发病过程.     

山茱萸总苷对关节炎大鼠原代滑膜细胞培养上清趋化能力的影响

探讨山茱萸总苷抗类风湿关节炎的作用机理.采用改良的Boyden小室前导法检测CⅡ诱导的关节炎大鼠山茱萸总苷治疗组和模型组的原代滑膜细胞培养上清对关节浸润炎性细胞如中性粒细胞、骨髓巨噬细胞和淋巴细胞趋化能力,结果以实验组穿膜细胞数减去对照组穿膜细胞数表示.结果发现:山茱萸总苷治疗组的原代滑膜细胞培养上清对上述三种炎性细胞的趋化能力(穿膜细胞数)均显著低于模型组(P＜0.01).说明山茱萸总苷抑制关节滑膜细胞对炎性细胞的趋化,减少炎性细胞向关节滑膜的浸润是其抗关节炎作用的机理之一.     

Paeoniflorin suppresses inflammatory mediator production and regulates G protein-coupled signaling in fibroblast – like synoviocytes of collagen induced arthritic rats

OBJECTIVE: To investigate the effects of Paeoniflorin (Pae) on inflammatory mediators and G protein - coupled signaling in fibroblast - like synoviocytes (FLS) from collagen induced arthritic (CIA) rats. METHODS: SD rats were injected with type II collagen. Pae (25, 50, 100 mg. kg(-1)) was administered to CIA rats. The inflammation of CIA rats was evaluated by paw swelling, arthritis index and histopathology of joints. FLS were isolated and cultured. Interleukin (IL)-1 activity was measured by the (3)H-TdR - intake method Tumor necrosis factor alpha (TNF-alpha), prostaglandin E(2) (PGE(2)) and cAMP were measured by radioimmunoassay. Protein kinase A (PKA) was assessed by luminescent kinase assay. Gi was detected by Western blot. RESULTS: Inflammation in CIA rats was accompanied by hyperplastic synovium, pannus and cartilage erosion in joints. IL-1 activity and Gi expression increased, PGE(2) and TNF-alpha production were enhanced, but cAMP level and PKA activity decreased. Pae significantly suppressed the inflammatory response and inflammatory mediators (IL-1, TNF-alpha and PGE(2)) in vivo. Pae inhibited Gi expression and restored cAMP level and PKA activity in FLS of CIA rats in vivo and vitro. CONCLUSION: Inflammatory mediators and G protein - coupled signaling were associated with the pathogenesis of synovitis in CIA rats. Pae, as a new monomer, had anti-inflammatory effects on the animal model of CIA in rats, but also had regulatory effects on FLS from CIA rats in vitro. These results highlight Pae as a good candidate for therapeutic intervention in RA.     

LncRNA PVT1 induces chondrocyte apoptosis through upregulation of TNF-α in synoviocytes by sponging miR-211-3p

Temporomandibular joint osteoarthritis (TMJ OA) is an important subtype of temporomandibular disorders (TMD). Articular cartilage destruction is considered a common pathological feature of TMJ OA, which is reported to be mainly induced by chondrocyte apoptosis. Synovial sterile inflammation is an initial factor of TMJ OA-associated articular cartilage destruction. Therefore, determining the mechanism of synovial membrane inflammation-induced articular cartilage destruction in TMJ OA is important for the TMJ OA therapy. In this study, we detected the function of synoviocytes in chondrocyte apoptosis under lipopolysaccharide (LPS)-induced inflammatory conditions and explored the underlying mechanism. We found that synoviocytes in inflammatory conditions facilitated LPS-induced chondrocytes apoptosis by secreting increased Tumor Necrosis Factor α (TNF-α), which was induced by long non-coding RNA plasmacytoma variant translocation 1 (PVT1) upregulation. PVT1 served as a competing endogenous RNA that sponged the microRNA miR-211-3p and prevented the inhibition of TNF-α expression. In conclusion, our in vitro study revealed that PVT1 has a previously unknown role in chondrocyte apoptosis, which may also be a mechanism underlying synoviocyte involvement in TMJ OA.     

热痹清片诱导AA大鼠滑膜细胞凋亡的作用及机制

目的：探讨热痹清片治疗类风湿关节炎（RA）的作用机制。方法：采用弗氏完全佐剂造成大鼠（AA）模型,设正常组、模型组、甲氨喋呤（MTX）组、雷公藤多苷（TPT）组和热痹清片组。分别采用电子显微镜、免疫组化法和流式细胞术检测各组大鼠滑膜细胞凋亡情况。结果：电镜观察显示,热痹清片组可见滑膜细胞核染色质呈凝块状,边聚呈月芽状,胞浆内及胞膜外可见凋亡小体;免疫组化法观察显示,热痹清片组Fas和FasL表达显著增加、Bcl-2表达显著减少（P〈0.05）,指标变化差异与TPT、MTX相当（P〉0.05）。流式细胞术观察表明,热痹清片组滑膜细胞凋亡率显著高于TPT组及MTX组（P〈0.05）。结论：热痹清片通过促进Fas、FasL表达,抑制Bcl-2的表达,促进了滑膜细胞凋亡,从而发挥治疗作用。