Combination benefit of a pyrimidylpiperazine derivative (Y-40138) and methotrexate in arthritic rats

Anti-tumor necrosis factor-alpha (TNFalpha) antibody in combination with methotrexate dramatically decreases joint destruction in rheumatoid arthritis. The aim of this study was to examine combined treatment with N-[1-(4-([4-(pyrimidin-2-yl)piperazin-1-yl]methyl)phenyl)cyclopropyl] acetamide HCl (Y-40138) and methotrexate in rat adjuvant-induced arthritis. The increase in hindpaw volume and joint destruction was suppressed by single therapeutic administration (days 15-20) of Y-40138 (30 mg/kg, p.o.), but not by prophylactic administration (days 1-9). However, arthritic progression was suppressed by single prophylactic administration of methotrexate (0.3 mg/kg, p.o.), but not by therapeutic administration. Combined administration (days 10-20) of Y-40138 (0.3-1 mg/kg) and methotrexate (0.03 mg/kg) synergistically suppressed the increase in hindpaw volume and joint destruction. We concluded that Y-40138 in combination with methotrexate synergistically suppressed arthritic progression. These data suggest that combined treatment with Y-40138 and methotrexate may increase efficacy of therapy for rheumatoid arthritis.     

Weekly pulse therapy of methotrexate improves survival compared with its daily administration in MRL/lpr mice

The purpose of this study was to determine whether weekly pulse therapy was superior to daily administration of methotrexate in MRL/lpr mice. Oral methotrexate was given to 6-week-old MRL/lpr mice at doses of 0.3, 1.0, or 3.0 mg/kg 5 days a week or at a dose of 15.0 mg/kg once a week until 35 weeks of age. The effects of methotrexate on physical, serological, and pathological findings were assessed. The survival rate and articular destruction on X-ray films were also evaluated. Both weekly pulse therapy and daily administration of methotrexate at the same weekly dose improved nephropathy and articular destruction of MRL/lpr mice when compared with control. However, weekly pulse therapy with methotrexate prolonged the survival of MRL/lpr mice when compared with the daily administration of the same weekly dose of methotrexate and control. Methotrexate did not suppress the increase in anti-DNA antibody and rheumatoid factor. Daily administration of methotrexate reduced the red and white blood cell counts, whereas weekly pulse therapy caused little reduction. In conclusion, weekly pulse therapy was superior to daily administration of methotrexate with respect to the survival rate, possibly due to a reduction in toxicity.     

重组人内抑素对佐剂性关节炎大鼠关节软骨酸敏感离子通道的影响及其保护作用

目的观察重组人内抑素(rh-End)对佐剂性关节炎大鼠关节软骨酸敏感离子通道(ASICs)表达的影响,探讨其对关节软骨的作用。方法将大鼠随机分成正常组、AA模型组、rh-End 1.25、2.5、5.0mg·kg-1组和阿司匹林50mg·kg-1阳性对照组。弗氏完全佐剂(CFA)致佐剂性关节炎(adjuvant-induced arthritis,AA)后d10,大鼠出现继发性炎症,此时皮下注射重组人内抑素,连续7d,对照组灌胃给阿司匹林;正常组与模型组给予等容量的无菌注射用水。光学显微镜观察关节病理变化,半定量RT-PCR方法检测rh-End对AA大鼠关节软骨ASICs和蛋白聚糖体mRNA的影响,Western blot方法检测rh-End对AA大鼠关节软骨ASICs蛋白的影响,免疫组织化学技术测定rh-End对AA大鼠关节软骨中的Ⅱ型胶原蛋白合成的影响。结果rh-End各剂量组能明显抑制AA大鼠关节软骨组织中的ASICs表达,并升高关节软骨基质成分Ⅱ型胶原蛋白和蛋白聚糖体mRNA表达量。结论重组人内抑素通过抑制ASICs的表达进而保护AA大鼠关节软骨。     

Therapeutic effects of Hominis placenta injection into an acupuncture point on the inflammatory responses in subchondral bone region of adjuvant-induced polyarthritic rat

Rheumatoid arthritis (RA) is characterized by chronic inflammation of the synovial membrane in the joint, which leads to the progressive destruction of articular cartilage, ligament and bone. Several cytokines such as tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta), and interleukin 6 (IL-6) have been implicated in the pathological mechanisms of synovial tissue proliferation, joint destruction and programmed cell death in rheumatoid joint. In the Korean traditional medicine, Hominis placenta (HP) as an herbal component of herb-acupuncture has been widely used to treat chronic inflammatory diseases such as RA. To study the therapeutic effects of HP injection into the ST36 acupoint (HP herb-acupuncture) on the inflammatory responses of a subchondral region of rheumatoid joint, the polyarthritis-induced Sprague-Dawley (SD) rat was developed as a rheumatoid arthritis model by the intradermal injection of dried cells of Mycobacterium tuberculosis emulsified in squalene to the base of tail. After the onset stage (11 d after adjuvant injection) of polyarthritis, a fixed volume of HP extract was daily injected to Zusanli (ST36) acupoint on the rat's leg for 2 weeks. The body weight, paw volume of the knee joint and articular index were exploited as an assessment method addressing arthritic symptoms, and the expression profiles of TNF-alpha, IL-1beta and IL-6 at the subchondral bone of the joint were analyzed using an immunohistochemistry. After the treatment of arthritic rats with HP, the body weights and paw volumes of arthritic rats were almost restored to the levels of normal rats whereas the evaluation by the articular index was not remarkable. The TNF-alpha, IL-1beta and IL-6 positive cells in the immunohistological sections of subchondral bone region of the joint significantly decreased in HP-treated (ST36 acupoint) arthritic group as compared with those in non-treated or HP-treated (non-acupoint) ones, which was coincident with the behavioral studies. In conclusion, the HP herb-acupuncture was found to be effective to alleviate the arthritic symptoms in adjuvant-induced arthritis rats as regards the body weight, joint appearance and the expression profiles of inflammatory cytokines.     

Suppression of adjuvant arthritis of rats by a novel matrix metalloproteinase-inhibitor

BAY 12-9566 (4-[4-(chlorophenyl)phenyl]-4-oxo-2S-(phenylthiomethyl) butanoic acid) is a newly developed, synthetic matrix metalloproteinase (MMP) inhibitor (MMPI) that selectively inhibits MMP-2, MMP-3 and MMP-9 isozymes. We study the effect of BAY 12-9566 on inflammation and cartilage destruction in adjuvant-induced arthritis (AA) in rats. Rats were injected with adjuvant and treated for 21 days with vehicle, Indomethacin or BAY 12-9566. AA was assessed: by measuring arthritic index, paw volume, urinary pyridinoline (Pyr) and deoxypyridinoline (Dpyr); by examining joint inflammation; and by microscopic morphometry of articular cartilages. Oral treatment of rats for 22 days with 50 mg kg(-1) body weight/d BAY 12-9566 showed decreased AA as determined by improvement in body weight gain (P<0.01), arthritic index (P<0.05) and swelling of paws contralateral to the adjuvant injection site (P<0.05). Neutrophil infiltration and collagen degradation were also significantly lower (P<0.01) in this treatment group. Cartilage destruction was successfully suppressed (P<0.01) in rats treated with either 50 mg kg(-1) body weight/d BAY 12-9566 or 1 mg kg(-1) body weight/d Indomethacin. These results indicate that BAY 12-9566 successfully suppressed inflammation and cartilage destruction in rats with AA. Moreover, these results also suggested that MMP-2, MMP-3 and MMP-9 are involved in arthritic diseases such as rheumatoid arthritis.     

Effects of leflunomide on human cartilage

Modern therapeutic approach in rheumatoid arthritis (RA) includes early use of disease-modifying anti-rheumatic drugs (DMARDs). DMARDs may influence the course of disease progression, and their introduction in early RA is recommended to limit irreversible joint damage. Among DMARDs, leflunomide and methotrexate are more utilised in pharmacological therapy. In the present work, we considered the effects of leflunomide, in comparison with those of methotrexate and to those of leflunomide-methotrexate combination on human cartilage to verify its effectiveness in arthritic disease, simulated by our experimental model. We measured in vitro the amount of glycosaminoglycans (GAGs) and the production of nitric oxide (NO) released into the culture medium of human articular cartilage treated with interleukin-1beta (IL-1beta), which promotes the cartilage destruction during articular disease. Leflunomide, in the presence of IL-1beta decreased NO production and GAGs release respect IL-1beta alone treated samples, in dose-related manner. Our results suggest that leflunomide is able to protect cartilage matrix from degradative factors induced by IL-1beta with respect to methotrexate and leflunomide-methotrexate combination.     

几种抗类风湿性关节炎药物对佐剂性关节炎大鼠关节软骨细胞酸敏感离子通道的表达影响及作用

目的：观察几种抗类风湿性关节炎（RA）药物对佐剂性关节炎（AA）大鼠关节软骨中的酸敏感离子通道（ASICs）表达的影响，探讨抗RA药物对关节软骨的保护作用。方法：将大鼠随机分成正常组、AA模型组、阿司匹林（50mg／kg）组、地塞米松（0．2mg／kg）组、雷公藤多苷（50mg／kg）组和左旋咪唑（10mg／kg）组。弗氏完全佐剂（CFA）致炎后第10天起，AA大鼠出现继发性炎症，此时地塞米松组腹腔注射地塞米松，其它用药组分别灌胃相应的受试药物，正常组与模型组灌胃等容量的无菌注射用水，连续7d。实验结束后，光学显微镜观察关节病理变化；用半定量RT-PCR和westem blotting分析常用药物对AA大鼠关节软骨细胞中ASICs表达的影响；用半定量RT-PCR方法分析常用药物对AA大鼠关节软骨中的蛋白聚糖体（aggregate proteoglycan，aggrecan）表达的影响；免疫组织化学技术检测常用药物对AA大鼠关节软骨中的Ⅱ型胶原蛋白合成的影响。结果：经过治疗后，各用药组能明显抑制AA大鼠的继发性足肿胀。阿司匹林能显著抑制AA大鼠关节软骨细胞ASICs的表达，其它用药组对ASICs无明显抑制作用。各用药组能明显升高AA大鼠关节软骨细胞基质成分Ⅱ型胶原和aggrean表达量，其中阿司匹林作用显著。结论：阿司匹林可能通过抑制ASICs的表达而抑制AA大鼠关节软骨的破坏。     

Methotrexate use in rheumatoid arthritis. A Clinician's perspective

Aminopterine, a precursor of methotrexate (MTX), was first used for the treatment of rheumatoid arthritis (RA) in 1951 [Gubner, R., 1951. Therapeutic suppression of tissue reactivity: I. Comparison of the effects of cortisone and aminopterin. Am. J. Med. Sci. 221, 169-175; Gubner, R., August, S., Ginsberg, V., 1951. Therapeutic suppression of tissue reactivity: II. Effect of aminopterin in rheumatoid arthritis and psoriasis. Am. J. Med. Sci. 221, 176-182.]. Corticosteroids, and to some extent cyclophosphamide, took MTX out of the rheumatologist's armamentarium until the late 1970s-early 1980s when the toxic profile of these compounds became apparent. By the mid 1980s, four randomized clinical trials (RCTs) had proven beyond doubt the beneficial effects of MTX when administered to patients with established disease who had failed to respond to other compounds such as gold salts and D-penicillamine [Thompson, R.N., Watts, C., Edelman, J., Esdaile, J., and Russell, A.S., 1984. A controlled two-centre trial of parenteral methotrexate therapy for refractory rheumatoid arthritis. J. Rheumatol. 11, 760-763; Andersen, P.A., West, S.G., O'Dell, J.R., Via, C.S., Claypool, R.G., and Kotzin, B.L., 1985. Weekly pulse methotrexate in rheumatoid arthritis. Clinical and immunologic effects in a randomized, double-blind study. Ann. Intern. Med. 103, 489-496; Weinblatt, M.E., Coblyn, J.S., Fox, D.A., Fraser, P.A., Holdsworth, D.E., Glass, D.N., and Trentham, D.E., 1985. Efficacy of low-dose methotrexate in rheumatoid arthritis. N. Engl. J. Med. 312, 818-822; Williams, H.J., Willkens, R.F., Samuelson, C.O.J., Alarcón, G.S., Guttadauria, M., Yarboro, C., Polisson, R.P., Weiner, S.R., Luggen, M.E., Billingsley, L.M., Dahl, S.L., Egger, M.J., Reading, J.C., and Ward, J.R., 1985. Comparison of low-dose oral pulse methotrexate and placebo in the treatment of rheumatoid arthritis. A controlled clinical trial. Arthritis Rheum. 28, 721-730.]. Subsequently, these four trials were included in a meta-analysis and the drug was approved by the Food and Drug Administration for use in RA [Health and Public Policy Committee, H.P.P.C. and American College Physicians, A.C.P., 1987. Methotrexate in rheumatoid arthritis. Ann. Intern. Med. 107, 418-419; Paulus, H.E., 1986. FDA Arthritis Advisory Committee meeting: Methotrexate; guidelines for the clinical evaluation of antiinflammatory drugs; DMSO in scleroderma. Arthritis Rheum. 29, 1289-1290; Tugwell, P., Bennett, K., and Gent, M., 1987. Methotrexate in rheumatoid arthritis. Indications, contraindications, efficacy, and safety. Ann. Intern. Med. 107, 358-366.]. Since then, rheumatologists have become aware of what Pincus et al. have called "the side effects" of RA comparing the morbidity and mortality caused by RA with that potentially caused by medications used to treat this disease [Pincus, T. and Callahan, L.F., 1993. The "side effects" of rheumatoid arthritis: joint destruction, disability and early mortality. Br. J. Rheumatol. 32, 28-37.]. Thus, during the 1990s the use of MTX for the treatment of RA became generalized [O'Dell, J.R., 1997. Methotrexate use in rheumatoid arthritis. Rheum. Dis. Clin. N Am. 23, 779-796 (a); Bannwarth, B., Vernhes, J., Schaeverbeke, T., and Dehais, J., 1995. The facts about methotrexate in rheumatoid arthritis. Rev. Rhum. 62, 471-473 (b); Bologna, C., Jorgensen, C., and Sany, J., 1997a. Methotrexate as the initial second-line disease modifying agent in the treatment of rheumatoid arthritis patients. Clin. Exp. Rheumatol. 15, 597-601; Bologna, C., Viu, P. (ABSTRACT TRUNCATED)     

Evaluation of the effect of losartan and methotrexate combined therapy in adjuvant-induced arthritis in rats

There is increasing body of evidence documenting the involvement of angiotensin II in inflammatory diseases. Moreover the up-regulation of angiotensin II AT₁ receptors in the synovium of rheumatoid arthritis patients has been previously described.This study aimed at investigating the anti-inflammatory effect of losartan, the selective angiotensin II AT₁ receptor blocker, and comparing the efficacy of methotrexate alone and in combination with losartan in adjuvant arthritis in rats. Twelve days post adjuvant injection, Sprague-Dawley rats were treated with methotrexate (1 mg/kg/week), losartan (20 mg/kg/day) and their combination for 15 days. Severity of arthritis was assessed by hind paw swelling, arthrogram scores. Serum was analyzed for measurement of albumin, C-reactive protein (CRP), nitrite/nitrate concentrations, interleukin 1β (IL-1β), tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), aspartate transaminase (AST) and alanine transaminase (ALT). Histopathological examination was done for hind paws and livers. Methotrexate and losartan monotherapies significantly reduced all parameters of inflammation and arthritis with better results in the methotrexate group except for the transaminases where losartan caused more significant reduction in their serum levels. The combined therapy showed better results than methotrexate and losartan alone. Hind paws showed better improvement of inflammatory cell infiltration and bone resorption in the combined therapy group. Disturbances in liver architecture and fibrosis caused by adjuvant arthritis were reverted to normal status in the combined therapy group in contrast to losartan and methotrexate monotherapies. In conclusion, methotrexate and losartan combined therapy provided more effective anti-inflammatory and hepatoprotective effects than either drug alone.     

Dual effect of nitric oxide donor on adjuvant arthritis

The effect of medical use of NO donors on the pathogenesis of arthritis is still yet unclear. We investigated the effects of the NO donor, sodium nitroprusside (SNP), on the pathogenesis of adjuvant-induced arthritis in rats. Rats were given SNP intraperitoneally either from day 5 to day 14 (as a prophylactic protocol) or from day 16 to day 25 (as a therapeutic protocol) after inoculation of adjuvant. SNP administration, whether prophylactic or therapeutic, in doses of 0.1 and 1 mg/kg/d significantly aggravated pathogenesis of adjuvant arthritis in rats. SNP-treated rats showed significant (P < 0.05) increase in arthritis index, hind paw volume, ankle joint diameter and hyperalgesia compared with control adjuvant arthritic rats. However, in adjuvant rats given the smallest dose of SNP (0.01 mg/kg/d), arthritis index, volume of hind paws, ankle joint diameter, body weight loss, and hyperalgesia were significantly lower than that of control adjuvant rats. After 30 d of the induction of adjuvant arthritis, TNF alpha levels exhibited insignificant changes either in control adjuvant rats or in rats given SNP compared with control non adjuvant rats. IL-10 levels in adjuvant control rats and adjuvant rats given 1 mg or 0.1 mg/kg/d from day 15 to day 25 were significantly lower than that of control non adjuvant rats. Histopathology examination of ankle joint showed that large doses of SNP (1 mg or 0.1 mg/kg/d) increased the mononuclear cells infiltration and erosion of cartilage induced by adjuvant while the infiltration of the inflammatory cells in the synovium of adjuvant rats treated with 0.01 mg/kg/d was minimal and the pannus was inhibited with alleviation of erosion of articular cartilage. Prophylactic small dose of SNP improved the histological status more than the therapeutic small dose. The present work reveals that SNP administration, either prophylactic or therapeutic, was deleterious in higher doses. However, the smallest dose used 0.01 mg/kg/d attenuates joint inflammation, hyperalgesia and body weight loss in adjuvant arthritic rats. These results suggest that small dose of NO donor may exert partial protective effects while the safety of the clinical use of NO donors, in higher doses, in patients with rheumatoid arthritis is questioned.     

Selective cyclooxygenase-2 (COX-2) inhibitors reduce anti-Mycobacterium antibodies in adjuvant arthritic rats

Adjuvant arthritis, induced by Mycobacterium butyricum, is an experimental immunopathy that shares many features of human rheumatoid arthritis and, as such, is one of the most widely used models for studying the anti-inflammatory activity of compounds. In rats with adjuvant induced arthritis, IgG antibodies to M. butyricum have been detected and autoantigens that cross react with mycobacteria may be involved in the pathogenesis of adjuvant arthritis. In this study, the anti-inflammatory and immunosuppressive activities of two cyclooxygenase-2 selective inhibitors, flosulide and L-745,337, at doses of 0.1, 1 and 5 mg/kg/day, were examined in adjuvant arthritic rats. After 14 days of treatment, a clear dose-dependent inhibition of plantar edema was seen for both flosulide (ID50 lower than 0.1 mg/kg) and L-745,337 (ID50 = 0.4 mg/kg). Plasma levels of IgG anti-M. butyricum antibodies were also decreased by both drugs. In each case the maximal immunosuppressive effect was observed at doses lower than 5 mg/kg. The non-selective COX-2 inhibitor, indomethacin (1 mg/kg) decreased paw edema by 65% and the levels of IgG anti-M. butyricum by 45%. Neither cyclooxygenase selective inhibitors nor indomethacin decreased the delayed hypersensitivity reaction induced by M. butyricum. Thus, in vivo inhibition of COX-2 inhibited articular swelling and also the humoral immune response to Mycobacterium.     

The effect of tryptophan plus methionine, 5-azacytidine, and methotrexate on adjuvant arthritis of rat.

Within the wider framework of our studies on the genesis of rheumatoid arthritis we have investigated the two signal processes in arthritis: adenoribosylation of proteins and DNA methylation. Arthritis can be induced when Freund's complete adjuvant is applied to rats. This form of arthritis can then be reduced or even totally suppressed through the application of several different substances. In the present article we have investigated if the effect of two of these substances, 5-azacytidine and methotrexate can be influenced by the application of tryptophan plus methionine. When applied singly, these latter two substances are known to reduce the formation of arthritis. This effect is intensified by a combination of tryptophan plus methionine. Application of tryptophan plus methionine without methotrexate or 5-azacytidine causes an enhanced development of an adjuvant induced arthritis.     

Cinnamaldehyde suppresses NLRP3 derived IL-1β via activating succinate/HIF-1 in rheumatoid arthritis rats

Cinnamaldehyde (CA) is an essential component of cinnamon (Cinnamomum cassia Presland), which is often used as a flavoring condiment in beverages, pastries, perfumes, etc. Cinnamon is also used as herbal medicine in China and Southeast Asia to treat rheumatoid arthritis. However, the molecular mechanism is unclear. In this study, we aim to investigate its anti-inflammatory effects against Rheumatoid arthritis (RA) using activated macrophages (Raw246.7) in vitro and adjuvant arthritis rats (AA) in vivo. The results demonstrated that CA significantly reduced synovial inflammation in AA rats, possibly due to suppression of the expressions of pro-inflammatory cytokines, especially the IL-1β. Further investigation found that CA also suppressed the activity of HIF-1α by inhibiting the accumulation of succinate in cytoplasm. As we know, the reduction of HIF-1α nucleation slows down IL-1β production, because HIF-1α activates the expression of NLRP3, which is involved in the assembly of inflammasome and processing of IL-1β. In addition, CA also inhibited the expression of the succinate receptor GPR91, which in turn inhibited the activation of HIF-1α. In conclusions, our results suggested that CA might be a potential therapeutic compound to relieve rheumatoid arthritis progress by suppressing IL-1β through modulating succinate/HIF-1α axis and inhibition of NLRP3.     

Age-depending effects of methotrexate treatment on systemic bone turnover in experimental adjuvant arthritis

Adjuvant arthritis was induced in rats in the growth stage (aged 6 weeks) and those in the mature stage (aged 4 months), and changes in the systemic bone turnover and the effects of methotrexate (MTX, CAS 133073-73-1) were compared. After induction of adjuvant arthritis, the paw edema ratio and the urinary deoxypyridinoline (u-Dpy) level increased in both age groups. No marked changes were observed in the serum osteocalcin (s-OC) level in either group. In the 6-week-old rats, arthritis completely inhibited the bone mass, and strength of the femur and lumbar vertebral body. The 4-month-old rats showed more marked changes than the 6-week-old rats in the bone mass and strength of the lumbar, vertebral body. MTX administration (0.05, 0.1 and 0.2 mg/kg/day) resulted in significant dose-dependent inhibition of arthritis-induced changes, and the effects of MTX were similar between the two age groups. MTX was useful at each age. These results suggest that 4-month-old rats with arthritis are more appropriate as a model for evaluation of drugs for bone metabolic turnover in human chronic rheumatoid arthritis.     

Effect of celecoxib, a cyclooxygenase-2 inhibitor, on the pathophysiology of adjuvant arthritis in rat

We investigated the efficacy of celecoxib, a specific cyclooxygenase (COX)-2 inhibitor, on arthritic pathophysiology and confirmed its gastric safety in adjuvant-induced arthritis rats. Results were compared with those for loxoprofen, a non-selective COX inhibitor. Arthritis was induced by injection of 1 mg of Mycobacterium butyricum in 50 microl of liquid paraffin into the left footpad of Lewis rats. The drugs were given by twice daily oral administration for 10 days beginning 15 days after adjuvant injection, with celecoxib at 0.01-3 mg/kg/day and loxoprofen at 0.01-3 mg/kg/day. Celecoxib significantly inhibited paw swelling, hyperalgesic response, and joint destruction (radiographic and histopathological findings) in these arthritic rats. These effects of celecoxib were superior to those of loxoprofen. Further, the administration of loxoprofen (3 mg/kg/day) caused significant gastric lesions, whereas celecoxib at the same dose did not. These results suggest that COX-2-mediated prostaglandins may play an important role in the progression of pathophysiology in this model and that celecoxib may be a useful therapeutic agent for the treatment of rheumatoid arthritis, with greater safety than non-selective COX inhibitors.     

紫草素对大鼠佐剂性关节炎滑膜炎症的影响

目的：探讨紫草素对佐剂性关节炎大鼠滑膜炎症的影响。方法：SD大鼠分为4组（每组10只）：正常对照组,佐剂性关节炎组,甲氨蝶呤治疗组,紫草素治疗组。测量大鼠后肢关节肿胀程度,实验第二十一天杀死大鼠,分离右后肢关节滑膜,应用免疫组织化学方法结合病理学图像分析系统,研究滑膜组织中单核-巨噬细胞（CCR5阳性）的浸润情况。结果：甲氨蝶呤和紫草素均能减轻关节肿胀（P〈0.05）。甲氨蝶呤治疗组和紫草素治疗组关节滑膜CCR5阳性细胞明显减少,且紫草素治疗组更低。结论：紫草素减轻大鼠佐剂性关节炎的炎症反应,减轻关节肿胀和减少滑膜组织炎症细胞浸润,为紫草素治疗类风湿关节炎（RA）提供了实验依据。     

Effects of ulinastatin on experimental arthritis

Effects of the urinary enzyme inhibitor ulinastatin on experimental arthritis were investigated. Ulinastatin at the dose of 30,000 units/kg/day, i.v., significantly restored the swelling of hind paw, inflammatory score and bone damage in adjuvant arthritic rats. Intraarticular administration of ulinastatin at 3000 units/site x 3, significantly suppressed the articular swelling and the elevated inflammatory parameters in the synovial fluid of carrageenin-induced arthritic rabbits. Moreover, ulinastatin at the dose of 50000 units/kg/day, i.v., significantly prevented the elevation of serum rheumatoid factor and articular lesions in MRL/l mice. In addition, ulinastatin significantly inhibited human granulocyte elastase and cathepsin G. These findings indicate that ulinastatin may be a useful therapeutic agent for arthritis.     

Anti-arthritic activity of synthesized chondroitin sulfate E hexasaccharide

The purpose of this study was to investigate the anti-arthritic effects of synthesized chondroitin sulfate E hexasaccharide (sCSE-6, CAS 866407-73-0), using a type II collagen-induced arthritis model in mice. sCSE-6 was administered subcutaneously on a daily basis to type II collagen (CII)-sensitized mice from day 0 to day 55. The severity of arthritis, as well as the immunohistological features of the arthritic mice, were analyzed. sCSE-6 inhibited the course of arthritis and restored the body weight loss of CII-immunized mice. An immunohistological analysis showed that bone/cartilage destruction in the arthritic mice was significantly attenuated by sCSE-6 treatment, with a marginal inhibition of synovial inflammation also observed. The beneficial effect of sCSE-6 on bone destruction, which is the most important factor in preventing arthritis, is particularly noteworthy. In summary, sCSE-6 inhibited arthritis and helped to prevent bone and cartilage destruction in a type II collagen-induced arthritis model in mice. The findings indicated that CSE oligosaccharides might be a novel potential therapeutic tool for rheumatoid arthritis.     

小剂量激素联合来氟米特与甲氨蝶呤治疗类风湿关节炎50例

目的探讨小剂量激素联合来氟米特与甲氨蝶呤治疗类风湿关节炎的临床疗效。方法将类风湿关节炎患者100例随机均分为治疗组和对照组,两组均给予甲氨蝶呤10mg/周和来氟米特10mg/d。治疗组加用强的松10mg,1次/d,疗程48周,病情控制后逐渐减量至停药,一般为3～6个月。治疗12周后观察两组患者晨僵、关节肿胀数、关节压痛数、双手握力、血沉（ESR）等指标。结果治疗组总有效率为88.00%,明显高于对照组的70.00%;治疗组各项观察指标均较对照组有显著变化（P〈0.05）。结论小剂量激素联合来氟米特与甲氨蝶呤治疗类风湿关节炎疗效满意。     

Therapeutic index of methotrexate depends on circadian cycling of tumour necrosis factor‐α in collagen‐induced arthritic rats and mice

Objectives Rheumatoid arthritis is an autoimmune disorder of unknown aetiology. Morning stiffness, a characteristic feature of rheumatoid arthritis, shows a 24‐h rhythm. Noticing this rhythm, we hypothesized the presence of a similar rhythm for a rheumatoid arthritis indicator, in addition to dosing‐time dependency of the anti‐rheumatic effect of methotrexate in arthritis induced by collagen in rats and mice, which reflect the symptomatology of rheumatoid arthritis patients. Methods To measure tumour necrosis factor (TNF)‐α concentration, blood was taken at different times (2, 6, 10, 14, 18 or 22 h after the light was turned on (HALO)) in collagen‐induced arthritic mice. Methotrexate was administered at two different dosing times based on these findings to estimate arthritis. Key findings The arthritis score was significantly lower in the 22 HALO‐treated group than in the control and 10 HALO‐treated groups in collagen‐induced arthritic rats and mice. Plasma TNF‐α concentrations showed obvious 24‐h rhythms, with higher levels at light phase and lower levels at dark phase after rheumatoid arthritis crisis. Arthritis was relieved after administration of methotrexate during the dark phase in synchronization with the 24‐h rhythm. Conclusions Our findings suggest that choosing an optimal dosing time associated with the 24‐h cycling of TNF‐α could lead to effective treatment of rheumatoid arthritis by methotrexate.