Studies on type II collagen induced arthritis in mice

A consistent and reproducible polyarthritis was induced in mice by immunizing them with type II collagen in Complete Freunds adjuvant (CFA) and Bacillus Calmette-Guerin (BCG) vaccine. Several inbred strains of mice were investigated for the ability to develop collagen induced arthritis (CIA). DBA/1 mice (H-2q) produced the highest incidence and the most severe arthritis of all the strains examined. Viable BCG vaccine was essential for the induction of a reproducible disease in this strain. The effects of some anti-inflammatory and anti-rheumatic compounds were examined on the developing and established lesions of CIA. These effects were determined by assessing the paw inflammation using a subjective scoring system and measuring foot weight. Furthermore, levels of serum amyloid P component (SAP) were also determined. Benoxaprofen, cyclophosphamide, indomethacin and prednisolone inhibited the paw inflammation in the developing disease whilst the anti-rheumatic compounds auranofin and D-penicillamine exacerbate the paw inflammation. Cyclophosphamide and prednisolone inhibited the established lesions but only prednisolone prevented the development of further lesions in the established disease. The SAP levels in the prednisolone treated group were also reduced. Auranofin treatment exacerbated the inflammation of both the established and the developing lesions in the same animal. D-penicillamine was inactive in the established disease.     

口服鸡Ⅱ型胶原治疗大鼠的佐剂型关节炎

目的 研究口服鸡Ⅱ型胶原对大鼠佐剂型关节炎的治疗作用。方法 建立大鼠佐剂型关节炎动物模型，通过胃管给药方式分别给予高、中、低剂量的鸡Ⅱ型胶原，对大鼠四肢关节进行评分，观察大鼠关节病变的发生和严重程度，采用免疫荧光技术，用流式细胞仪检测大鼠外周血T细胞亚群的变化，并且用大鼠rINFn检测试剂盒(固相夹心ELISA法)检测大鼠外周血TNFα水平的变化。结果 建立了大鼠佐剂型关节炎实验动物模型，口服高、中、低剂量的鸡Ⅱ型胶原均可减轻关节病变的发生，其中高剂量治疗组的作用最显著，而且大鼠外周血CD3 T细胞、CD4 T细胞和CD8 T细胞水平与关节炎模型组比较均有显著性差异，外周血TNFα水平也较模型组显著降低。结论 口服鸡Ⅱ型胶原可有效减轻实验性佐剂型关节炎大鼠的关节病变，其机理与口服耐受有关。     

可溶性鸡Ⅱ型胶原对关节炎大鼠的免疫治疗作用

目的 :考察口服小剂量可溶性鸡Ⅱ型胶原(SCCⅡ )对大鼠佐剂性关节炎 (AA)的免疫治疗作用。方法 :检测脾细胞、肠系膜淋巴结细胞 (MLNCs)和肠派伊尔结细胞 (PPCs)的ConA增殖反应、腹腔巨噬细胞 (PMΦ)和滑膜细胞 (SMCs)产生IL 1和TNFα水平 ,结合观察踝关节继发炎症、胸腺指数等指标变化。结果 :SCCⅡ (0 .0 3、0 .30和 3.0 0mg·kg- 1·d- 1,ig)治疗 (d 1 2～1 8) ,能提升患鼠脾细胞低下的ConA增殖反应 ,进一步抑制MLNCs和PPCs低下的ConA增殖反应 ,显著抑制PMΦ和SMCs产生IL 1和TNFα,明显减轻大鼠AA的炎症反应。结论 :口服小剂量可溶性SCCⅡ对大鼠AA有免疫治疗作用     

Humoral and cellular immunologic aspects of adjuvant and collagen arthritis in rats

Systemic and local immunological responses of rats sensitized with either M. butyricum or native type II collagen have been evaluated. In rats exhibiting adjuvant-induced arthritis no antibodies to collagen could be detected. In animals exhibiting collagen-induced arthritis, high antibody titers developed by day 14, and could be correlated with the severity of the arthritis. Delayed type hypersensitivity (DTH) responses were measured by a 5-iodo-2'-deoxyuridine 125-I (125-IUdR) uptake assay. Arthritic scores in rats immunized with collagen were not accompanied by a positive DTH response, whereas adjuvant arthritic rats showed a positive response. T-lymphocyte cellular responses in both adjuvant- and collagen-induced arthritic rats were measured. In neither syndrome were major alterations observed in T-lymphocyte subpopulations. These results provide evidence that adjuvant-induced arthritis and type II collagen-induced arthritis are distinct entities, and that they may be discriminated by the nature of the humoral response.     

Differences in pharmacokinetics and hepatobiliary transport of a novel anti-inflammatory agent between normal and adjuvant arthritis rats

1. The pharmacokinetics, particularly the hepatobiliary transport of T-5557 ((3-methyl-2-oxo-piperadin-3-yl)-acetic acid N'-(3-thieophen-2-yl-8-methoxy-quinazolin-1-yl)-hydrazide), a novel anti-inflammatory agent, has been examined in normal and adjuvant arthritis (AA) rats. 2. Following oral administration of T-5557, the absolute bioavailability in AA rats was increased by sixfold compared with normal rats. The extent of binding T-5557 to plasma proteins obtained from AA rats was markedly greater than in normal rats (97.0 versus 88.2%). The biliary clearance in AA rats was significantly lower than that in normal rats (1.186 versus 5.621 ml min(-1) kg(-1)), and lower intrinsic biliary clearance was also observed in AA rats (40.33 versus 69.83 ml min(-1) kg(-1)). 3. Concomitant administration of T-5557 with quinidine, a potent P-glycoprotein inhibitor, to normal rats caused a significant decrease in the biliary clearance of T-5557 by 37.9%. Moreover, the transport of T-5557 for the apical-to-basal compartment in a Caco-2 cells' monolayer was fourfold lower than that for the opposite direction, and was increased in the presence of quinidine and verapamil. 4. These results suggest that P-glycoprotein is involved in the biliary excretion of T-5557 and the decrease in the transport activity as well as the increase in plasma protein binding caused the elevated plasma concentration and bioavailability of T-5557 in AA rats.     

Differences in pharmacokinetics and hepatobiliary transport of a novel anti-inflammatory agent between normal and adjuvant arthritis rats

1. The pharmacokinetics, particularly the hepatobiliary transport of T-5557 ((3-methyl-2-oxo-piperadin-3-yl)-acetic acid N '-(3-thieophen-2-yl-8-methoxy-quinazolin-1-yl)-hydrazide), a novel anti-inflammatory agent, has been examined in normal and adjuvant arthritis (AA) rats. 2. Following oral administration of T-5557, the absolute bioavailability in AA rats was increased by sixfold compared with normal rats. The extent of binding T-5557 to plasma proteins obtained from AA rats was markedly greater than in normal rats (97.0 versus 88.2%). The biliary clearance in AA rats was significantly lower than that in normal rats (1.186 versus 5.621 ml?min -1 kg -1), and lower intrinsic biliary clearance was also observed in AA rats (40.33 versus 69.83 ml?min -1 kg -1) . 3. Concomitant administration of T-5557 with quinidine, a potent P-glycoprotein inhibitor, to normal rats caused a significant decrease in the biliary clearance of T-5557 by 37.9%. Moreover, the transport of T-5557 for the apical-to-basal compartment in a Caco-2 cells' monolayer was fourfold lower than that for the opposite direction, and was increased in the presence of quinidine and verapamil. 4. These results suggest that P-glycoprotein is involved in the biliary excretion of T-5557 and the decrease in the transport activity as well as the increase in plasma protein binding caused the elevated plasma concentration and bioavailability of T-5557 in AA rats.     

霞水母胶原治疗大鼠佐剂型关节炎的研究

目的 探讨霞水母来源的胶原对大鼠佐剂型关节炎的治疗作用.方法 采用完全弗氏佐剂造成大鼠关节炎模型,致炎后大鼠给予霞水母胶原连续灌胃14 d,观察大鼠双后肢关节肿胀情况,及对血清中一氧化氮(NO),超氧化物歧化酶(SOD)、丙二醛(MDA)的影响.结果 口服霞水母胶原可以降低大鼠的双后肢关节肿胀度,降低大鼠血清中NO和MDA含量,提高大鼠血清中SOD活性.结论 霞水母胶原对佐剂型关节炎有较好的治疗作用,其机理可能与降低体内氧自由基水平及过氧化脂质水平有关.     

鸡Ⅱ型胶原对小鼠胶原性关节炎的治疗作用

目的　研究鸡Ⅱ型胶原 (CⅡ )对小鼠胶原性关节炎(CIA)的治疗作用。方法　在建立小鼠CIA模型的基础上 ,观测关节积分变化 ,同时检测脾淋巴细胞增殖反应 ,白细胞介素 1(IL 1)和IL 2。结果　CⅡ 5、10、2 0 μg·kg-1ig× 7d能明显减轻CIA小鼠的关节炎症 ;抑制CIA小鼠过高的ConA诱导的脾细胞增殖反应及小鼠腹腔巨噬细胞IL 1的过度产生 ,降低脾细胞IL 2的产生 ;同时病理检查亦发现CⅡ治疗用药可以减轻CIA小鼠的骨膜增生和炎细胞浸润。结论　CⅡ对CIA具有治疗作用 ,其机制与免疫功能调节有关。     

Adjuvant arthritis in young copper-deficient rats.

Adjuvant arthritis has been studied in young copper-deficient rats and a very strong inhibition of the disease following 60 days of deficient diet (0.4 ppm of copper) was found. No difference in lung prostaglandin synthesis and in rat stomach strip and colon reactivity to exogenous prostaglandins was noticed between controls and copper-deficient animals. These results are briefly discussed.     

可溶性鸡Ⅱ型胶原对大鼠胶原性关节炎的免疫治疗作用

目的　考察可溶性鸡Ⅱ型胶原 (SCCⅡ )对大鼠胶原性关节炎 (CIA)的免疫治疗作用及机制。方法　检测CIA大鼠血清抗Ⅱ型胶原 (CⅡ )抗体、腹腔巨噬细胞 (PMΦ)与滑膜细胞产生白介素 1(IL 1)和肿瘤坏死因子α(TNFα)和SCCⅡ /刀豆球蛋白A(ConA)诱导 5个部位淋巴细胞的增殖反应 ,同时测量继发炎症反应和免疫器官系数。结果　ig 0 .0 3,0 .3和 3.0mg·kg- 1·d- 1SCCⅡ (d 14～ 2 2 ) ,明显减轻患鼠继发炎症 ,抑制针对SCCⅡ的皮肤迟发型超敏反应和PMΦ,滑膜细胞超常分泌IL 1和TNFα ,促进体重和免疫器官系数恢复 ,提升低下的淋巴细胞ConA反应 ,但不能降低血清抗CⅡ抗体水平 ,5个部位的淋巴细胞对SCCⅡ体外刺激也几无反应性。结论　SCCⅡ通过T细胞免疫耐受途径对大鼠CIA产生治疗作用。     

Phellinus baumii ethyl acetate extract alleviated collagen type II induced arthritis in DBA/1 mice

Mushrooms have a long history of dietary benefits in Asia due to their health-promoting effects. Phellinus baumii, a wild mushroom, has been reported to have anti-platelet, anti-inflammatory, anti-obesity and free radical scavenging activities. However, its anti-rheumatoid arthritis (RA) property remains poorly understood. Hence, we investigated the protective effect of Phellinus baumii ethyl acetate extract (PBEAE) against bovine collagen type II induced arthritis (CIA) in DBA/1 mice. PBEAE (50 and 150 mg/kg) reduced the CIA score and leukocyte count in draining lymph nodes (DLNs) and inflamed joints. PBEAE also attenuated the expressions of CD3⁺ (T cells), CD19⁺ (B cells), CD4⁺ (T-helper), CD8⁺ (T-cytotoxic), MHC class II/CD11c⁺ (antigen-presenting cells), double positives (B220⁺/CD23⁺ and CD3⁺/CD69⁺: early lymphocyte activation markers) and CD4⁺/CD25⁺ (activated T-helper) leukocyte subpopulations in DLNs. Likewise, CD3⁺ and Gr-1⁺CD11b⁺ (neutrophil) counts in inflamed joints were also decreased. Furthermore, PBEAE reduced the serum levels of anti-collagen type immunoglobulin G, tumor necrosis factor-α and interleukin (IL)-1β and IL-6. Taken together, PBEAE impaired cellular recruitment to the inflamed joint and alleviated CIA, and thus could be considered as a potential agent against rheumatoid arthritis     

Paradoxical effects of LS-2616 (Linomide) treatment in the type II collagen arthritis model in mice

The quinoline-3-carboxamide, Linomide, has been shown to possess potent immunomodulatory activity. We have evaluated the effect of Linomide in the type II collagen induced arthritis in mice. Treatment with Linomide (1.25-80 mg/kg/day) from the day of immunization strongly suppressed the arthritic response. On the other hand, initiation of treatment (20-80 mg/kg/day) at the onset of arthritis resulted in an increased severity of the arthritis. These potent and contradictory effects of Linomide, depending on treatment regime, indicates that central immuno-regulatory functions are affected and that this compound may be a useful tool for the understanding of autoimmune mechanisms.     

佐剂性关节炎(AA)大鼠血清中瘦素(leptin)与TNF-α的关系

目的:研究瘦素(leptin)在佐剂性关节炎(AA)大鼠血清中的表达,以及与TNF-α的关系,以探讨leptin在类风湿关节炎(RA)发病机制中的作用。方法:通过给大鼠右后足足跖部皮下注射完全弗氏佐剂(CFA)建立类风湿关节炎(RA)的动物模型-AA。用放免法(RIA)测定AA大鼠血清中leptin,TNF-α的表达水平。结果:AA组大鼠血清中leptin水平高于正常组,TNF-α水平高于正常组,且血清中leptin的表达水平与TNF-α呈显著正相关。结论:AA大鼠血清中leptin水平增高,并与TNF-α呈显著正相关,leptin可能参与了RA的发病,为RA的治疗提供了新的途径。     

Safety and toxicological evaluation of undenatured type II collagen

Previous research has shown that undenatured type II collagen is effective in the treatment of arthritis. The present study evaluated the broad-spectrum safety of UC-II by a variety of toxicological assays including acute oral, acute dermal, primary dermal irritation, and primary eye irritation toxicity. In addition, genotoxicity studies such as Ames bacterial reverse mutation assay and mouse lymphoma tests, as well as a dose-dependent 90-day sub-chronic toxicity study were conducted. Safety studies indicated that acute oral LD₅₀ of UC-II was greater than 5000?mg/kg in female Sprague-Dawley rats. No changes in body weight or adverse effects were observed following necropsy. Acute dermal LD₅₀ of UC-II was determined to be greater than 2000?mg/kg. Primary skin irritation tests conducted on New Zealand Albino rabbits classified UC-II as slightly irritating. Primary eye irritation tests conducted on rabbits indicated that UC-II was moderately irritating to the eye. UC-II did not induce mutagenicity in the bacterial reverse mutation test in five Salmonella typhimurium strains either with or without metabolic activation. Similarly, UC-II did not induce a mutagenic effect in the gene mutation test in mouse lymphoma cells either with or without metabolic activation. A dose-dependent 90-day sub-chronic toxicity study revealed no pathologically significant changes in selected organ weights individually or as percentages of body or brain weights. No significant changes were observed in hematology and clinical chemistry. Therefore, the results from the current study show a broad-spectrum safety profile of UC-II.     

Ⅱ型胶原诱导的小鼠关节炎动物模型的建立及影响因素

类风湿性关节炎(RA)是一种顽固性、致畸性自身免疫性疾病。Ⅱ型胶原诱导的小鼠关节炎(CIA)表现为严重的多关节性炎,并能引起慢性、破坏性关节损伤,与RA有很多相似之处,目前国际上广泛用于RA发病机制的研究以及创新药物的研发。该文就CIA实验动物模型的建立以及CIA发生病变的主要影响因素进行综述。