薯蓣皂苷对大鼠胶原性关节炎治疗作用及其机制研究
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摘要
类风湿关节炎(rheumatoid arthritis, RA)是一种慢性、系统性、自身免疫性疾病,其主要病理特点是滑膜细胞增生,衬里层增厚,多种炎性细胞浸润,血管翳形成,继而引起软骨和骨组织的破坏,最终导致关节畸形和功能丧失。目前治疗RA的药物可分为非甾体抗炎药、甾体抗炎药和慢作用抗风湿药。由于RA的病因和发病机制仍不甚明了,临床应用受到限制。寻找安全、有效、毒副作用小的治疗RA的中药是今后研究的重要方向之一。中药穿山龙(Dioscorea nipponica Makino),主要成分有穿山龙含薯蓣皂甙(dioscin),纤细薯蓣皂甙(gracillin),穗菝葜甾甙(asperin),25-D-螺甾-3,5-二烯(25-D-spirosta-3,5-diene)及对羟基苄基酒石酸(piscidic acid)。中医作药物为其根茎,有舒筋活络,祛风止痛之功效。大鼠胶原诱导性关节炎(collagen-induced arthritis, CIA)是兼有体液免疫和细胞免疫变化的慢性系统性免疫性炎症,与RA有许多共同的组织学和免疫学特征,是筛选和研究治疗RA药物的较理想模型。本课题在建立大鼠CIA模型的基础上,从整体、器官、细胞、分子、信使RNA基因表达等不同层次研究了Dioscin对CIA大鼠的抗炎和免疫调节作用,并对其部分机制进行了探讨,为开发和利用穿山龙资源,提供理论依据。
1Dioscin体内给药对大鼠CIA炎症和免疫反应的影响
应用异源性鸡Ⅱ型胶原(CCⅡ)和弗氏不完全佐剂(FIA)建立大鼠CIA模型,用足爪仪测量足爪肿胀度(Δml),同时采用O-4级评分法进行关节评分；检测体重变化情况(Ag)；测定大鼠胸腺指数和脾指数；用ELISA法检测CIA大鼠抗CCⅡ抗体；用螺旋测微计测量两耳厚度之差(Δmm)观察CIA大鼠对CCⅡ的迟发型变态反应；用H&E染色的方法观察病理组织学改变；结果发现CIA大鼠炎症和免疫反应表现为足爪肿胀和多发性关节炎；体重和免疫器官(胸腺和脾)减轻；体内产生抗CⅡ抗体和对CCⅡ的迟发型变态反应；其病理组织学显示炎性细胞浸润、滑膜增厚、关节软骨破坏,与人RA类似,Dioscin30,60,120mg·kg-1,(ig)×7d均能明显减轻CIA大鼠足爪肿胀和多发性关节炎,缓解体重和免疫器官(胸腺和脾)重量的异常变化；对CIA大鼠抗CCⅡ抗体无明显作用；可显著抑制CIA大鼠的迟发性变态反应；Dioscin可明显减轻异常的病理组织学变化,对CIA大鼠病变关节的病理组织结构有明显的改善作用。
2Dioscin体内、体外给药对CIA大鼠免疫细胞和细胞因子的作用
采用MTT法检测胸腺、脾淋巴细胞增殖反应；用活化的小鼠脾淋巴细胞MTT比色法检测胸腺淋巴细胞产生的IL-2的活性；采用ConA诱导的小鼠胸腺淋巴细胞MTT参入法检测腹腔巨噬细胞产生的IL-1的水平；采用生物学方法检测腹腔巨噬细胞产生的TNF-α的活性；结果表明CIA大鼠ConA诱导的胸腺T细胞增殖反应和IL-2的产生显著增高,LPS诱导的脾B细胞增殖反应无明显变化：腹腔巨噬细胞产生的IL-1和TNF-α的活性明显升高。Dioscin30、60、120mg·kg-1,(ig)×7d和体外0.5、2.5、12.5、62.5、125mg·L-1不同浓度可抑制CIA大鼠过高的胸腺T细胞增殖反应和IL-2的产生；对于腹腔巨噬细胞IL-1、TNF-α的过度产生也有抑制作用。
3Dioscin体内、体外给药对CIA大鼠滑膜细胞增殖及其分泌功能的影响
RA早期的病变位于滑膜,其滑膜细胞功能的改变直接影响RA病理进程。原代培养细胞离体时间短,其生物学特性和遗传性基本与体内相似,无自发性变异,是细胞形态、机能、分化等研究的理想材料。本文采用MTT法检测滑膜细胞的增殖反应：采用ConA诱导的小鼠胸腺淋巴细胞MTT参入法检测滑膜细胞产生的IL-1的水平；采用生物学方法检测滑膜细胞产生的TNF-α的活性；结果表明CIA大鼠滑膜细胞的增殖反应升高；滑膜细胞产生的IL-1和TNF-α的活性明显升高。Dioscin30、60、120mg·kg1,(ig)×7d和体外0.5、2.5、12.5、62.5、125mg·L-1不同浓度可抑制CIA大鼠过高的滑膜细胞的增殖反应；对于滑膜细胞IL-1、TNF-α的过度产生也有抑制作用。
4Dioscin对胶原诱导性大鼠关节炎踝关节滑膜组织中NF-κBp65亚基和COX-2表达的影响
为了探讨Dioscin对CIA大鼠炎症免疫反应的作用及其机制,本研究采用Western-blog方法检测CIA大鼠滑膜组织中NF-κBp65亚基和COX-2的表达。研究发现,CIA大鼠滑膜细胞的NF-κBp65亚基和COX-2的表达水平明显高于正常对照组,Dioscin120mg·kgl能显著降低关节炎大鼠踝关节滑膜组织中NF-κBp65亚基和COX-2表达(P<0.01),Dioscin(60mg/kg)和吲哚美辛亦可降低COX-2表达(P<0.05)。提示：GCS通过抑制NF-κBp65亚基和COX-2表达,从而抑制滑膜细胞的增殖反应和细胞因子及炎症介质的分泌而发挥抗炎和免疫作用。
综上所述,本文采用体内外研究相结合从整体、器官、细胞、分子、基因表达等不同层次探讨了Dioscin对大鼠CIA的作用及其NF-κBp65亚基和COX-2表达的关系,结果表明Dioscin对大鼠CIA具有治疗作用,其机制与调节机体的免疫功能、抑制炎症性细胞因子的过度分泌和调节NF-κBp65亚基和COX-2表达有关。调节NF-κBp65亚基和COX-2表达可能是GCS发挥抗炎免疫作用的重要分子机制之一。
Rheumatoid arthritis (RA) is an autoimmune disease that expresses chronic inflammation in synovial tissues and joints and develops into impaired joint function. The prevalence of RA varies worldwide between0.5%and1%. Genetic, hormonal, and environmental factors contribute to its development. A major obstacle to the development of rational treatment strategies is that the disease mechanisms and the causative environmental and genetic factors remain largely unknown. Clues may come from experimental arthritis.
Dioscorea nipponica Makino is a Chinese tradltional herbal medieine(CTM). Effective parts and chemical constituents of Dioscorea nipponica Makino includes dioscin, gracillin, asperin,25-D-spirosta-3,5-diene and piscidic acid. Dioscin was extracted and purified by methods of solvent extraction,column chromatography, layer chromatography, et al.
Collagen-induced arthritis (CIA) is a well-known disease model for RA. CIA resembles RA in a number of pathological, histological and immunological aspects. Features of CIA include chronic synovitis,inflammatory cell infiltration, Pannus formation,destruction of cartilage, and bone erosion. Immune mechanisms that include both humoral and cellular immunity to CⅡ have been implicated in the pathogenesis of the disease. Furthermore, the similarities between the joint pathology in CIA and RA are most widely used for studies of RA pathogenesis and for screening of new drugs for treatment of the rheumatoid disease. This study delineates the disease course, the influence of the organ-and tissue-specificity of inflammation, and the dynamics of the joint inflammation including infiltrating cell types, the influence of T cells, the humoral and cellular reactivity to the arthritogenic cartilage autoantigens rat collagen typeⅡ(CⅡ).The goal of this study was to extend our understanding of the roles played by Dioscin in the pathogenesis of arthritic diseases by interfering with cytokine-signaling pathways. Our study also aimed to develop a new therapeutic strategy for the treatment of RA by blockade of intracellular cytokine-signaling pathways.
1.Effccts of Dioscin on inflammatory and immune responses of collagen-induecd arthritis in vivo
CIA model was induced by native chicken type Ⅱ collagen that had been dissolved overnight at4℃in0.1M acetic acid(4mg·mL-1) and emulsified with an equal volume of Freund's incomplete adjuvant(FIA). Hindpaw volumes of rats were measured by volume meter(△ml) and arthritic index was evaluated at the same time; weight gain and arthritic index of immune organ(thymus and spleen) were also examined; antibodies to CⅡ was determined by ELISA and the delayed-type hypersensitivity(DTH)(△mm)was also examined; The legs and hindpaws of rats were removed, fixed with10%Paraformaldehyde in PBS, and then decalcified for10days with EDTA and embedded in paraffin for histologic analysis. The paraffin sections were stained with hematoxylin and eosin;It was shown that the administration of Dioscin (30,60,120mg·kg-1, ig×7d) inhibited the inflammatory response and restored bodyweight and the weight of immune organs of CIA rats. Dioscin had no effect on the concentration of antibodies to CⅡ. It was found that there were increases of lymphocyte proliferation、IL-2production and synoviocytes proliferation in CIA rats, together with IL-1andTNF-a in peritoneal microphages.
Dioscin reduced above changes significantly and had good effect on the progressive inflammatory, degeneration of synovial, cartilage, and bone in arthritic joints of CIA rats.
2. Effects of Dioscin on immune cells and cytokines of collagen-induced arthritis in vivo and vitro
ConA and LPS-induced lymphocytes proliferation, the production of interleukin-2(IL-2) secreted by thymus, the production of interleukin-1(IL-1) secreted by peritoneal macrophages(PMφ) were determined by MTT assay; TNF-α level of PMφ were determined by quantitation of the cytotoxic activity of TNF-a on L929cells respectively. Results showed that the increases of lymphocyte proliferation and IL-2production in CIA rats could be inhibited by Dioscin30,60,120mg·kg-1, ig×7din vivo and with0.5,2.5,12.5,62.5,125mg·L-1invtro, the same results as that of IL-1and TNF-α in PMφ.
3. Effects of Dioscin on synovial cells and cytokines of collagen-induced arthritis in vivo and vitro
Synovial cells were prepared by collagenase and trypsin digestion of small minced membranes. Synoviocytes proliferation and IL-1levels were determined by MTT assay; TNF-α level of synovial cells were determined by quantitation of the cytotoxic activity of TNF-α on L929cells, respectively. Results showed that the increases of synovial cells proliferation in CIA rats could be inhibited by Dioscin at the administration of30,60,120mg·kg-1, igx7d in vivo and at concentration of o.5,2.5,12.5,62.5,125mg·L-1in vitro, the same results as that of IL-1and TNF-a in synovial cells.
4. Effects of Dioscin on the expressions of NF-κBp65subunit and COX-2in rats' synovium of ankle joint
The expressions of NF-κBp65subunit and COX-2in rats' synovium of ankle joint were determined by Western blot method. we found that NF-κBp65subunit and COX-2expression in CIA rats was more high than that of normal, which could be inhibited by Dioscin at the administration of60,120mg·kg-1igx7d, Results showed that Dioscin could lower the levels of NF-KBp65subunit and COX-2in rats' synovium of ankle joint.
1Dioscin体内给药对大鼠CIA炎症和免疫反应的影响
应用异源性鸡Ⅱ型胶原(CCⅡ)和弗氏不完全佐剂(FIA)建立大鼠CIA模型,用足爪仪测量足爪肿胀度(Δml),同时采用O-4级评分法进行关节评分；检测体重变化情况(Ag)；测定大鼠胸腺指数和脾指数；用ELISA法检测CIA大鼠抗CCⅡ抗体；用螺旋测微计测量两耳厚度之差(Δmm)观察CIA大鼠对CCⅡ的迟发型变态反应；用H&E染色的方法观察病理组织学改变；结果发现CIA大鼠炎症和免疫反应表现为足爪肿胀和多发性关节炎；体重和免疫器官(胸腺和脾)减轻；体内产生抗CⅡ抗体和对CCⅡ的迟发型变态反应；其病理组织学显示炎性细胞浸润、滑膜增厚、关节软骨破坏,与人RA类似,Dioscin30,60,120mg·kg-1,(ig)×7d均能明显减轻CIA大鼠足爪肿胀和多发性关节炎,缓解体重和免疫器官(胸腺和脾)重量的异常变化；对CIA大鼠抗CCⅡ抗体无明显作用；可显著抑制CIA大鼠的迟发性变态反应；Dioscin可明显减轻异常的病理组织学变化,对CIA大鼠病变关节的病理组织结构有明显的改善作用。
2Dioscin体内、体外给药对CIA大鼠免疫细胞和细胞因子的作用
采用MTT法检测胸腺、脾淋巴细胞增殖反应；用活化的小鼠脾淋巴细胞MTT比色法检测胸腺淋巴细胞产生的IL-2的活性；采用ConA诱导的小鼠胸腺淋巴细胞MTT参入法检测腹腔巨噬细胞产生的IL-1的水平；采用生物学方法检测腹腔巨噬细胞产生的TNF-α的活性；结果表明CIA大鼠ConA诱导的胸腺T细胞增殖反应和IL-2的产生显著增高,LPS诱导的脾B细胞增殖反应无明显变化：腹腔巨噬细胞产生的IL-1和TNF-α的活性明显升高。Dioscin30、60、120mg·kg-1,(ig)×7d和体外0.5、2.5、12.5、62.5、125mg·L-1不同浓度可抑制CIA大鼠过高的胸腺T细胞增殖反应和IL-2的产生；对于腹腔巨噬细胞IL-1、TNF-α的过度产生也有抑制作用。
3Dioscin体内、体外给药对CIA大鼠滑膜细胞增殖及其分泌功能的影响
RA早期的病变位于滑膜,其滑膜细胞功能的改变直接影响RA病理进程。原代培养细胞离体时间短,其生物学特性和遗传性基本与体内相似,无自发性变异,是细胞形态、机能、分化等研究的理想材料。本文采用MTT法检测滑膜细胞的增殖反应：采用ConA诱导的小鼠胸腺淋巴细胞MTT参入法检测滑膜细胞产生的IL-1的水平；采用生物学方法检测滑膜细胞产生的TNF-α的活性；结果表明CIA大鼠滑膜细胞的增殖反应升高；滑膜细胞产生的IL-1和TNF-α的活性明显升高。Dioscin30、60、120mg·kg1,(ig)×7d和体外0.5、2.5、12.5、62.5、125mg·L-1不同浓度可抑制CIA大鼠过高的滑膜细胞的增殖反应；对于滑膜细胞IL-1、TNF-α的过度产生也有抑制作用。
4Dioscin对胶原诱导性大鼠关节炎踝关节滑膜组织中NF-κBp65亚基和COX-2表达的影响
为了探讨Dioscin对CIA大鼠炎症免疫反应的作用及其机制,本研究采用Western-blog方法检测CIA大鼠滑膜组织中NF-κBp65亚基和COX-2的表达。研究发现,CIA大鼠滑膜细胞的NF-κBp65亚基和COX-2的表达水平明显高于正常对照组,Dioscin120mg·kgl能显著降低关节炎大鼠踝关节滑膜组织中NF-κBp65亚基和COX-2表达(P<0.01),Dioscin(60mg/kg)和吲哚美辛亦可降低COX-2表达(P<0.05)。提示：GCS通过抑制NF-κBp65亚基和COX-2表达,从而抑制滑膜细胞的增殖反应和细胞因子及炎症介质的分泌而发挥抗炎和免疫作用。
综上所述,本文采用体内外研究相结合从整体、器官、细胞、分子、基因表达等不同层次探讨了Dioscin对大鼠CIA的作用及其NF-κBp65亚基和COX-2表达的关系,结果表明Dioscin对大鼠CIA具有治疗作用,其机制与调节机体的免疫功能、抑制炎症性细胞因子的过度分泌和调节NF-κBp65亚基和COX-2表达有关。调节NF-κBp65亚基和COX-2表达可能是GCS发挥抗炎免疫作用的重要分子机制之一。
Rheumatoid arthritis (RA) is an autoimmune disease that expresses chronic inflammation in synovial tissues and joints and develops into impaired joint function. The prevalence of RA varies worldwide between0.5%and1%. Genetic, hormonal, and environmental factors contribute to its development. A major obstacle to the development of rational treatment strategies is that the disease mechanisms and the causative environmental and genetic factors remain largely unknown. Clues may come from experimental arthritis.
Dioscorea nipponica Makino is a Chinese tradltional herbal medieine(CTM). Effective parts and chemical constituents of Dioscorea nipponica Makino includes dioscin, gracillin, asperin,25-D-spirosta-3,5-diene and piscidic acid. Dioscin was extracted and purified by methods of solvent extraction,column chromatography, layer chromatography, et al.
Collagen-induced arthritis (CIA) is a well-known disease model for RA. CIA resembles RA in a number of pathological, histological and immunological aspects. Features of CIA include chronic synovitis,inflammatory cell infiltration, Pannus formation,destruction of cartilage, and bone erosion. Immune mechanisms that include both humoral and cellular immunity to CⅡ have been implicated in the pathogenesis of the disease. Furthermore, the similarities between the joint pathology in CIA and RA are most widely used for studies of RA pathogenesis and for screening of new drugs for treatment of the rheumatoid disease. This study delineates the disease course, the influence of the organ-and tissue-specificity of inflammation, and the dynamics of the joint inflammation including infiltrating cell types, the influence of T cells, the humoral and cellular reactivity to the arthritogenic cartilage autoantigens rat collagen typeⅡ(CⅡ).The goal of this study was to extend our understanding of the roles played by Dioscin in the pathogenesis of arthritic diseases by interfering with cytokine-signaling pathways. Our study also aimed to develop a new therapeutic strategy for the treatment of RA by blockade of intracellular cytokine-signaling pathways.
1.Effccts of Dioscin on inflammatory and immune responses of collagen-induecd arthritis in vivo
CIA model was induced by native chicken type Ⅱ collagen that had been dissolved overnight at4℃in0.1M acetic acid(4mg·mL-1) and emulsified with an equal volume of Freund's incomplete adjuvant(FIA). Hindpaw volumes of rats were measured by volume meter(△ml) and arthritic index was evaluated at the same time; weight gain and arthritic index of immune organ(thymus and spleen) were also examined; antibodies to CⅡ was determined by ELISA and the delayed-type hypersensitivity(DTH)(△mm)was also examined; The legs and hindpaws of rats were removed, fixed with10%Paraformaldehyde in PBS, and then decalcified for10days with EDTA and embedded in paraffin for histologic analysis. The paraffin sections were stained with hematoxylin and eosin;It was shown that the administration of Dioscin (30,60,120mg·kg-1, ig×7d) inhibited the inflammatory response and restored bodyweight and the weight of immune organs of CIA rats. Dioscin had no effect on the concentration of antibodies to CⅡ. It was found that there were increases of lymphocyte proliferation、IL-2production and synoviocytes proliferation in CIA rats, together with IL-1andTNF-a in peritoneal microphages.
Dioscin reduced above changes significantly and had good effect on the progressive inflammatory, degeneration of synovial, cartilage, and bone in arthritic joints of CIA rats.
2. Effects of Dioscin on immune cells and cytokines of collagen-induced arthritis in vivo and vitro
ConA and LPS-induced lymphocytes proliferation, the production of interleukin-2(IL-2) secreted by thymus, the production of interleukin-1(IL-1) secreted by peritoneal macrophages(PMφ) were determined by MTT assay; TNF-α level of PMφ were determined by quantitation of the cytotoxic activity of TNF-a on L929cells respectively. Results showed that the increases of lymphocyte proliferation and IL-2production in CIA rats could be inhibited by Dioscin30,60,120mg·kg-1, ig×7din vivo and with0.5,2.5,12.5,62.5,125mg·L-1invtro, the same results as that of IL-1and TNF-α in PMφ.
3. Effects of Dioscin on synovial cells and cytokines of collagen-induced arthritis in vivo and vitro
Synovial cells were prepared by collagenase and trypsin digestion of small minced membranes. Synoviocytes proliferation and IL-1levels were determined by MTT assay; TNF-α level of synovial cells were determined by quantitation of the cytotoxic activity of TNF-α on L929cells, respectively. Results showed that the increases of synovial cells proliferation in CIA rats could be inhibited by Dioscin at the administration of30,60,120mg·kg-1, igx7d in vivo and at concentration of o.5,2.5,12.5,62.5,125mg·L-1in vitro, the same results as that of IL-1and TNF-a in synovial cells.
4. Effects of Dioscin on the expressions of NF-κBp65subunit and COX-2in rats' synovium of ankle joint
The expressions of NF-κBp65subunit and COX-2in rats' synovium of ankle joint were determined by Western blot method. we found that NF-κBp65subunit and COX-2expression in CIA rats was more high than that of normal, which could be inhibited by Dioscin at the administration of60,120mg·kg-1igx7d, Results showed that Dioscin could lower the levels of NF-KBp65subunit and COX-2in rats' synovium of ankle joint.
引文
[1].Jinnin M, Ihn H, Yamane K, Asano Y、Yazawa N, Jhmaki K. Clinicalfeatures of Patients with systemic sclerosis accompanied by theumatoid arthritis. ClinExP Rheumatol,2003;21(1):91-4
[2].Maetzel A, Tugwell P, Boers M, Guillemin F, Coyle D, DrummondM, etal. Economic Evaluation of Programs or Interventions in the Management of Reumatoid Arthritis:Defining a Consensus-based Reference Case. JR Reumatol,2003;30(4):891-6
[3]. Kaplan MJ, McCune WJ. New evidence for vaseular disease in patients with early rheumatoid arthritis. Lancet,2003:361(9363):1068-9
[4].Gulan G, Ravlie-Gulan J, StrboN, Sotosek V Nemec B, Matovinovie D, etal. Systemic and local expression of Perforin in lymphocyte subsets in acute and chronic rheumatoid arthritis. J Rheumatol,2003;30(4):660-70
[5]. Hanley DA, Brown JP, Tenenhouse A, Olszynski WP, Ioannidis G, Berger C, etal. Assoeiations amnong disease conditions, bone mineral density, and Prevalent vertebral deformities in men and wemen 50 years of age and older:cross-sectional results from the Canadian Multicentre Osteoporosis Study. J Bone MinerRes,2003:18(4):784-90
[6].Tallia AF, Cardone DA. Diagnostic and therapeutic injection of the should region. Am Fam Physician,2003:67(6):1271-8
[7]. Duryea J, Dobbins JT 3rd, Lynch JA. Digital tomosynthesis of hand joints for arthritis assessment.Med Phys,2003:30(3):325-33
[8]. Gonzalez-Alvarol, Carrnona L, Balsa A, Sanmarti R, Belmonte MA, Tena X. Patterns of disease modifying antirheumatic drug use in a Spanish cohort of Patients with rheumatoid arthritis. J Rheumatol,2003:30(4):697-704
[9]. WANG XP, LU X. New progress of Dioscin biological activity research [J]International Journal of Traditional Chinese Medicine,2004 26(3):138-141.
[10]. Larsson E, Erlandsson Harris H, Lorentzen JC, Larsson A, Mans son B, Klareskog L, etal. Serum concentrations of cartilage oligomeric matrix protein, fibrinogen and hyaiuronan distinguish inflammation and cartilage destruction in experimental arthritis in rats. Rheumatology(Oxford), 2002:41(9):996-1000
[11]. Lu S, Carlsen S, Hansson AS, Holmdahl R. Immunization of rats with homologous type XI collagen leads to chronic and relapsing arthritis with different genetics and joint pathology than arthritis induced with homologous type Ⅱ collagen. J Autoimmun,2002;18(3):199-211
[12]. Romas E, Sims NA, Hards DK, Lindsay M, Quinn JW, Ryan PF, etal. Osteoprotegerin reduces osteoelast numbers and prevents bone erosion in Collagen-induecd arthritis. AmJPathol,2002:161(4):1419-27
[13].董文娟,梁秀军,翟泽玲,宋鸿儒等.穿山龙总皂苷对CIA大鼠滑膜血管新生的作用.[J].承德医学院学报,2010,27(4)：360-362
[14].于海荣,王济兴,陈建双等.穿山龙总皂苷含药血清对小鼠脾淋巴细胞增殖及IL-6产生影响的实验研究.[J].江苏中医药,2007,39(1)：57-58[15]. Remmers EF, Joe B, Griffiths MM, Dobbins DE, Dracheva SV, Hashirajnoto A, etal. Modulation of multiple experimental arthritis models by collagen-induced arthritis quantitative trait loci isolated in congenic rat lines:different effeets of non-major histocompatibility complex quantitative trait loci in males and females. Arthritis Rheum,2002; 46(8):2225-34
[16]. Bendele A, MeAbee T, Sennello G, Frazier J, ChliPala E, MeCabe D. Effieacy of sustained blood levels of interleukin-1 receptor antagonist in animal models of arthritis:comparison of efficacy in animal models with human clinical data. Arthritis Rheum,1999:42(3):498-506
[17]. Cuzzocrea S, MeDonald MC, Mota-FiliPe H, Mazzon E, Costantlno G, Britti D, etal. Beneficial effects of tempol, a membrane-permeable radical scavenger, in a rodent model of collagen-induced arthritis.Arthritis Rheum,2000:43(2):320-8
[18]. Mizuno M, Nishikawa K, Spiller OB, Morgan BP, Okada N, Okada H, etal. Membrane complement regulators Protect against the development of type II Collagen-induced arthritis in rats. Arthritis Rheum,2001: 44(10):2425-34
[19]. CatehPole B, Staines NA, Hamblin AS. Antigen Presentation of Type II collagen in rats. Clin ExP Immnunol,2001:125(3):478-84
[20]. Fjuisavva H. Nishimura T, Motohaga M. Effect of actarton type Ⅱ Collagen induced arthritis in mice. Arznein Forsh/Drug Res,1994:44(1):64-8
[21]. Lu S, Carlsen S, Hansson AS, Holmdahl R. Immunization of rats with homologous type Ⅺ collagen leads to chronic and relapsing arthritis with different genetics and joint Pathology than arthritis induced with homologous type Ⅱ collagen. J Autoimmun,2002;18(3):199-211
[22]. Han Z, Chang L, yamanishi Y, Karin M, Firestein GS. Joint damage and Inflammation in c-Jun N-tenminal kinase 2 knockout mice with passive murine Collagen-induced arthritis. Arthritis Rheum,2002:46(3):818-23
[23].魏伟,沈玉先,徐叔云.免疫细胞培养技术.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1421-22
[24].魏伟,沈玉先,徐叔云.淋巴细胞功能的测定.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1426-28
[25].沈玉先,魏伟,徐叔云.细胞因子的测定.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1438-42
[26]. Cusehieri J, Gourlay D, Gareial I, Jelaeie S, Maier RV. Hypertonic preconditioning inhibits macrophage responsiveness to endotoxin. J Immunol,2002:168 (3):1389-96
[27]. Kiemer AK, Muller C, Vollmar AM. Inhibition of LPS-induced nitric oxide and TNF-alpha production by alpha-liPoic acid in rat KuPffer eells and in RAW 264.7 murine macrophages. Immunol Cell Biol,2002:80(6):550-7
[28].Sugiyama T, Ishii S, Yamamoto J, Irie R, Saito K, Otuki T, etal. cDNA macroarray analysis of gene expression in synoviocytes stimulated with TNF alpha. FEBS Lett,2002;517(1-3):121-8
[29].Privat J, Oo A, Waterbury LD. Production of interleukin-6, but not interleukin-8, induced by TNF-alpha or IL-1 beta in human f ibroblast-like ynoviocyte in ercreases over cell passage. Proc West Pharmacol Soc, 2001:44:9-13
[30]. Migita K, Thnaka F,s Yamasakis, Shibatomi K, Ida H, Kawakami A, etal. Regulation of rheumatoid synoviocyte Proliferation by endogenous P53 induction. Clin ExP Immunol,2001:126(2):334-8
[31].Nakazawa M, Ishii H, Aono H, Takai M, Honda T, Aiatanl S, etal. Role of Noteh-1 intral cellular domain in activation of rheumatoid synovioeytes.ArthritisRheum,2001:44(7):1545-54
[32]. Trentham DE. Townes AS, Kang AH. Autoimmunity to type Ⅱ collagen:an Experimental model of arthritis. J Exp Med,1977:146:857-66
[33], Romas E, Bakharevski O, Hards DK, Kartsogiannis V, Quinn JM, Ryan PF, etal.Expression of osteoelast differentiation factor at sites of bone erosion in collagen-induced arthritis. Arthritis Rheum,2000;43(4):821-6
[34]. Kobayashi K, Nakashima A, Nagata H, Nakajima H,、Yamaguehi K, Sato S, etal. Anti-arthritic effects of KF20444, a new immunosuppressive compound inhibiting Dihydroorotate dehydrogenase, on rat collagen-inducedarthritis.Inflamm Res, 2001:50(1):24-31
[35]. Nawata K, EnokidaM,、Yamasaki D, Minamizaki T, Hagino H, Morio Y etal. Tensile Properties of rat anterior cruciate ligament in collagen induced arthritis. Ann Rheum Dis,2001:60(4):395-8
[36].Hoshino K, Hanyu T, Arai K, Takahashi HE.Mineraldensity and histomorphometric assessment of bone changes in the proximal tibia early after induction of type Ⅱ Collagen-induced arthritis in growing and mature rats. J Bone Miner Metab,2001;19(2):76-83
[37]. Cuzzoerea S, Mazzon E, Bevilaqua C, Costantlno G, Britti D, Mazzullo G, etal. Clorieromene, a coumarine derivative, protects against collagen-induced arthritis in Lewis rats. Br J phannacol,2000:131 (7):1399-407
[38]. Yamasaki D, Enokida M, Okano T, Hagino H, Teshima R. Effects of ovariectomy and estrogen replacement therapy on arthritis and bone mineral density in rats with collagen-induced arthritis. Bone,2001:28(6):634-40
[39]. Wernhoff P, Unger C, Bajiner E, Burkhardt H, Holmdahl R. Identification of conformnation-dependent epitopes and V gene selection in the B cell response to type Ⅱ collagen in the DA rat. Int Immunol,2001; 13(7):909-19
[40]. CatehPole B, Staines NA, Hamblin AS. Antigen Presentation of Type Ⅱ collagen in rats.Clin Exp Immunol,2001:125(3):478-84
[41]. Romas E, Bakharevski O, Hards DK, Kartsogiarmis V, Quinn JM, Ryan PF, etal. Expression of osteoelast differentiation factor at sites of bone erosion in collagen-induced arthritis. Arthritis Rheum,2000;43 (4):821-6
[42]. Cheon, Yu SJ, Yoo DH, Chae IJ, Song GG, Sohn J. Increased expression of Pro-inflammatory cytokines and metal loproteinase-1 by TGF-beta 1 in synovial fibroblasts from rheumatoid arthritis and normal individuals. Clin ExP Immunol,2002:127(3):547-52
[43]. Brinkman DM, ten Cate R,Vossen JM.Smeeet TJ,Kraan MC,Tak PP. Decrease in syovial cellularity and cytokine expression after autologous stem cell transplantation in patients with juvenile idiopathic arthritis. ArthritisRheum,2002:46(4):1121-3
[44]. Okazaki A, Koshino T, Saito T, Takagi T. Osseous tissue reaction around Hydroxyapatite block implanted into proximal metaphysic of tibia of rat with collagen-induced arthritis. Biomaterials,2000:21(5):483-7
[45]. He W, Pelletier JP, Martel-Pelletier J, Laufer S, Di Battista JA.Synthesis of interleukinlbeta, tumornecrosis factor-alpha.and interstitial collagenase(MMP-1)15 eicosanoid dependent in human osteo arthritis synovial membrane explants:inieractions with anti-inflammatory cytokines.J Rheumatol,2002;29(3):546-53
[46].Matsuura M, Imayoshi T, Okumoto T. Effect of FTY720, a novel immunosuppressant, on adjuvant-and collagen-induced arthritis in rats. Int J Immunopharmacol,2000:22(4):323-31.
[47].Du Z F,Li Z B,Sun C X,et al. Effects of Artesunate on TNF-a and IL-1 in the serum of rats with ajuvant arthritis[J].Guangdong Medical Journal,2010,31 (13):1637-40.
[48]. Xie S J,Song H R.The Experimental Study on Immunoregulatory Effects of Whole Saponin of Rhizoma D ioscreae Nipponicae on AA Rat[J]. LiaoNing Journal of Traditional Chinese Medicine,2007,34 (9):1323-1325
[49].Li Y,Song Y Y,Zhang H Q.Effect of Echinacoside on immune function and mitochondrial DNA relative content of aging mice[J]. Chin Pharmaco Bull,2010,26 (6):810-3.
[50].Yun CX,Yu XL,Wu G et al. Effect of Sangyehuang mixture on cellular immunological function in rats[J]. Lishizhen Medicine Materia Medica Research,2009,20(5):1155-7.
[51]. Trentham DE.Townes AS, Kang AH. Autoimmunity to type Ⅱ collagen:an experimental model of arthritis.J Exp Med,1977;146:857-66
[52].Nawata K,Enokida M,Yamasaki D,Minamizaki T,Hagino H,Morio Y, et al. Tensile properties of rat anterior cruciate ligament in collagen induced arthritis. Ann Rheum Dis,2001;60(4):395-8
[53].Cai H, Zheng Z L,Shang W,et al.The effect of curcumin on the immune organ weight and pathomorphology of adjuvant arthritis rats[J].JETCM,2009,18(4):594-5.
[54].Yang X H,Wu J,Guo L J. Optimization on extraction technology of saponin of chaenomeles speciosa and study on its anti-inflammatory immunologic mechanism to rat adjuvant arthritis[J].2010,25(5):547-9.
[55].Yi J F,Lu A P. Study on the Immunology of Triptolide and Dihydroartem-isinine Compound Treating CIA Rats[J].Journal of Jiangxi university of TCM,2008,20(6):49-51.
[56].GAO YX,LIANG XJ,DONG WJ,et al. Effects of Total Saponin from Rhizoma Dioscreae Nipponicae on NF-κB p65 Activity and STAT3 Protein Expression in Joint Synovium of CIA Rats[J]. Journal of China Medical University Vol.41 No.6 Jun.2012, 41(6):485-489
[57].WANG XP, LU X. New progress of Dioscin biological activity research [J]. International Journal of Traditional Chinese Medicine,2004,26(3):138-141.
[58].LI SP, HU XM,JIN ZM. Therapeutic Effect of Dioscin on rheumatoid arthritis [J]. Zhejiang clinical medicine,2008,11(10):628.
[59].YANG JC, CHEN ZX. Medical Molecular Biology.BeiJing:Chemical Industry Press,2003:1204.
[60].YAO L, LIU SM. Study on the Effective Ingredients of Rhizoma Dioscoreae Nipponicae on Experimental Acute Gouty Arthritis[J]. Chinese Archives of Traditional Chinese Medicine,2010,28(8):1724-1726
[61].Yang X H,Wu J,Guo L J. Optimization on extraction technology of saponin of chaenomeles speciosa and study on its anti-inflammatory immunologic mechanism to rat adjuvant arthritis[J].2010,25(5):547-9.
[62].Yi J F, Lu A P. Study on the Immunology of Triptolide and Dihydroartem-isinine Compound Treating CIA Rats[J] Journal of Jiangxi university of TCM,2008,20(6):49-51.
[63].GM.Campo, A.Avenoso, S.Campo, et al. Chondroitin-4-sulphate inhibitsNF-κB translocation and caspase activation in collagen-induced arthritis inmice. Osteoarthritis and Cartilage,2008,4:1.
[64].DING Q, YOU ZL. Effects of compound components from Panax Notoginseng on NF-κBP65 and TNF-a, IL-β in inflammation model of human endometrial cell. [J] Chinese Journal of Traditional Medical Science and Technology,2007,14(2):113.
[65].Anderson GD,KeysKL,De Ciechi PA, et al Combination therapies that inhibit cyclooxygenase-2 and leukotriene synthesis prevent disease inmurine collagen induced arthritis. Inflamm Res,2009,2:109.
[66].Noguchi M, Kimoto A, Sasamata M, et al Micro-CT imaging analysis for the effect of celecoxib, a cyclooxygenase-2 inhibitor, on inflammatory bone destruction in adjuvant arthritis rats. J Bone Miner Metab,2008,26:461.
[67].Lee HS, Lee CH, Tsai HC, et al Inhibition of cyclooxygenase 2 expression by diallyl sulfide on joint inflammation induced by urate crystal and IL-1β.OsteoarthritisCartilage,2009,1:91.
[68].Gabay C.Cytokine inhibitors in the treatment of rheumatoid arthritis. Expert OPin Biol Ther,2002:2(2):135-49
[69]. Schrnitz M L, Baeuerle P A. The p65 subunnit is responsible for the strong transcription activating potential of NF-κB. EMBO J,1991; 10(12):3805
[70]. MLienhard S, MarcosA, Patrick A. Transactivation domain 2 (TA2) of p65 NF-κB. J. Biol. Chem,1995; 270(26):15576
[71]. Llui's E, Julia IE, Alex RM, et al. I-κB and p65 regulate the cyto-plasmic shuttling of ruclear corepressors:Cross-talk between notch and NF-κB pathways. Molecular Biology of the Cells,2003;14:491
[72].宋震坤,陈文照.环氧化酶-2与骨关节炎[J].中国骨伤,2003,13(5)：314-315.
[73].吴尚洁,蔡颖,赵水平.阿托伐他汀降低脂多糖诱导下人肺上皮细胞COX-2的表达[J].中国医师杂志,2004,6(6)：768.
[74].邓渝林,陶霞,周静等.第二代COX-2抑制剂的研究进展[J].解放军药学学报,2005,21(3)：209.
[75].戴生明,韩星海,施冶青.两种选择性COX-2抑制剂治疗类风湿关节炎和骨关节炎的临床比较[J].中华风湿病学杂志,2000,4(6)：378.
[76].唐福林,张奉春,郑文洁等.双醋瑞因治疗膝骨关节炎的疗效与安全性多中心对照研究[J].中华风湿病学杂志,2005,9(7)：423.
[77].皮治兵,蒋宗滨.COX-2抑制剂临床研究新进展[J].中国疼痛医学杂志,2006,12(5)：297.
[1]van Boekel MA, Erik RV, van den Hoogen FH, et al. Autoantibody systems in rheumatoid arthritis:specificity, sensitivity and diagnostic value. Arthritis Res,2002,4:87-93.
[2]Egeland T, Munthe E. The role of the laboratory in rheumatology. Rheumatoid factors. Clin Rheum Dis 1983,9(1):135-60.
[3]MagsaamJ, Ferjenck P, Tenmpels M, et al. A new method for the detection of hidden IgM rheumatoid factor in patients with juvenile rheumatoid arthritis[J].J Rheumatol 1987,14(5):964-967.
[4]何菁,贾汝琳,韩蕾,等.隐性类风湿因子在类风湿关节炎诊断中的意义[J].中华风湿病杂志,2001,5：24-27·
[5]Hoet RM, van Venrooij WJ. The antiperinuclear factor and antikeratin antibodies in rheumatoid arthritis. In:Smolen JS, Kalden JR, Maini RN, editors. Rheumatoid arthritis. Berlin: Springer Verlag; 1992. p.299-318.
[6]]Young BJ, Mallya RK, Leslie RD, Clark CJ, Hamblin TJ. Antikeratin antibodies in rheumatoid arthritis. Br Med J 1979; 2:97-9.
[7]穆荣,孙晓云,栗占国.类风湿因子和抗胍氨酸多肽抗体联合检测在类风湿关节炎诊断中的意义[J].北京大学学报,2005,37(5)：498-500.
[8]Forslind K, AhlmenM, EberhardtK, et al. Prediction of radiological outcome in early rheumatoid arthritis in clinical practice:role of antibodies to citrullinated peptides (anti-CCP) [J]. Ann Rheum Dis,2004, 63(9):1090-1095.
[9]El-Gabalawy HS, Wilkins JA. Anti-Sa antibodies:prognostic and pathogenetic significance to rheumatoid arthritis. Arthritis Res Ther, 2004,6:86-9.
[10]Hayem G, Chazerain P, Combe B, et al. Anti-Sa antibody is an accurate diagnostic and prognostic marker in adult rheumatoid arthritis [J]. J Rheumatol,1999;26(1):7-13
[11]Goldbach-Mansky R, Lee J, McCoy A, et al. Rheumatoid arthritis associated autoantibodies in patients with synovitis of recent onset. Arthritis Res,2000,2:236-243.
[12]钱龙,曾庆馀,黄少弼,等.抗核周因子的检测及其在类风湿关节炎中的意义[J].中华风湿病学杂志,1999;3(2)：100-103
[13]Schellekens GA, Visser H, de Jong BAW, et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide[J]. Artritis Rheum,2000; 43(1):155-163
[14]Blass S, Union A, Raymackers J, et al. The stress protein BiP is overexpressed and is a major B and T cell target in rheumatoid arthritis. Arthritis Rheum,2001,44:761-771.
[15]田昕,毕黎琦,栗占国.抗瓜氨酸化Ⅱ型胶原抗体的检测及其与类风湿关节炎的相关性研究[J].中华医学杂志,2006,86(33)：2334.
[16]Burkhardt H, Sehnert B, Bockermann R, et al. Humoral immune response to citrullinated collagen type Ⅱ determinants in early rheumatoid arthritis [J].Eur J Imm uno,12005,35(5):1643-1652.
[17]Nielen MM, vander Horst AR, van Sehaardenbug D, et al. Antibodies to citrullinated human fibrinogen(ACF) have diagnostic and prognostic value in early arthritis. Ann Rheum Dis,2005,64:1199-1204.
[18]赵义,田听,栗占国.抗瓜氨酸化纤维蛋白原抗体的检测及其在类风湿关节炎中的临床意义.北京大学学报,2006,38：350.
[19]Bang H, Luthke K, Burmester GR, et al. Mutated citrullinated Vimentin as a candidate autoantigen for diagnosis and monitoring of Disease activity in Rheumatoid Arthritis. Ann Rheum Dis,2006,65:144.
[20]姜东林,孙钧铭,姜升阳,等.类风湿关节炎患者抗MCV、抗CCP抗体与RF诊断价值比较[J].临床检验杂志,2009,27(2)：137-138.
[1]van Boekel MA, Erik RV, van den Hoogen FH, et al. Autoantibody systems in rheumatoid arthritis:specificity, sensitivity and diagnostic value. Arthritis Res, 2002,4:87-93.
[2]何菁,贾汝琳,韩蕾,等.隐性类风湿因子在类风湿关节炎诊断中的意义[J].中华风湿病杂志,2001,5：24-27.
[3]穆荣,孙晓云,栗占国.类风湿因子和抗胍氨酸多肽抗体联合检测在类风湿关节炎诊断中的意义[J].北京大学学报,2005,37(5)：498-500.
[4]Forslind K, Ahlmen M, Eberhardt K, et al. Prediction of radiological outcome in early rheumatoid arthritis in clinical practice:role of antibodies to citrullinated peptides (anti-CCP) [J].Ann Rheum Dis,2004,63(9):1090-1095.
[5]E1-Gabalawy HS, Wilkins JA. Anti-Sa antibodies:prognostic and pathogenetic significance to rheumatoid arthritis. Arthritis Res Ther,2004,6:86-9.
[6]Goldbach-Mansky R, Lee J, McCoy A, et al. Rheumatoid arthritis associated autoantibodies in patients with synovitis of recent onset. Arthritis Res, 2000,2:236-243.
[7]钱龙,曾庆馀,黄少弼,等.抗核周因子的检测及其在类风湿关节炎中的意义[J].中华风湿病学杂志,1999：3(2)：100-103
[8]Schellekens GA, Visser H, de Jong BAW, et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide[J]. Artritis Rheum,2000; 43(1):155-163
[9]Blass S, Union A, Raymackers J, et al. The stress protein BiP is overexpressed and is a major B and T cell target in rheumatoid arthritis. Arthritis Rheum, 2001,44:761-771.
[10]田昕,毕黎琦,栗占国.抗瓜氨酸化Ⅱ型胶原抗体的检测及其与类风湿关节炎的相关性研究[J].中华医学杂志,2006,86(33)：2334.
[11]Burkhardt H, Sehnert B, Bockermann R, et al. Humoral immune response to citrullinated collagen typeⅡdeterminants in early rheumatoid arthritis [J].Eur J Immunol 2005,35(5):1643-1652.
[12]Nielen MM, vander Horst AR, van Sehaardenbug D, et al. Antibodies to citrullinated human fibrinogen(ACF) have diagnostic and prognostic value in early arthritis. Ann Rheum Dis,2005,64:1199-1204.
[13]赵义,田听,栗占国.抗瓜氨酸化纤维蛋白原抗体的检测及其在类风湿关节炎中的临床意义.北京大学学报,2006,38：350.
[14]Bang H, Luthke K, Burmester GR, et al. Mutated citrullinatedVimentin as a candidate autoantigen for diagnosis and monitoring of Disease activity in Rheumatoid Arthritis. Ann Rheum Dis,2006,65:144.
[15]姜东林,孙钧铭,姜升阳,等.类风湿关节炎患者抗MCV、抗CCP抗体与RF诊断价值比较[J].临床检验杂志,2009,27(2)：137-138.
[2].Maetzel A, Tugwell P, Boers M, Guillemin F, Coyle D, DrummondM, etal. Economic Evaluation of Programs or Interventions in the Management of Reumatoid Arthritis:Defining a Consensus-based Reference Case. JR Reumatol,2003;30(4):891-6
[3]. Kaplan MJ, McCune WJ. New evidence for vaseular disease in patients with early rheumatoid arthritis. Lancet,2003:361(9363):1068-9
[4].Gulan G, Ravlie-Gulan J, StrboN, Sotosek V Nemec B, Matovinovie D, etal. Systemic and local expression of Perforin in lymphocyte subsets in acute and chronic rheumatoid arthritis. J Rheumatol,2003;30(4):660-70
[5]. Hanley DA, Brown JP, Tenenhouse A, Olszynski WP, Ioannidis G, Berger C, etal. Assoeiations amnong disease conditions, bone mineral density, and Prevalent vertebral deformities in men and wemen 50 years of age and older:cross-sectional results from the Canadian Multicentre Osteoporosis Study. J Bone MinerRes,2003:18(4):784-90
[6].Tallia AF, Cardone DA. Diagnostic and therapeutic injection of the should region. Am Fam Physician,2003:67(6):1271-8
[7]. Duryea J, Dobbins JT 3rd, Lynch JA. Digital tomosynthesis of hand joints for arthritis assessment.Med Phys,2003:30(3):325-33
[8]. Gonzalez-Alvarol, Carrnona L, Balsa A, Sanmarti R, Belmonte MA, Tena X. Patterns of disease modifying antirheumatic drug use in a Spanish cohort of Patients with rheumatoid arthritis. J Rheumatol,2003:30(4):697-704
[9]. WANG XP, LU X. New progress of Dioscin biological activity research [J]International Journal of Traditional Chinese Medicine,2004 26(3):138-141.
[10]. Larsson E, Erlandsson Harris H, Lorentzen JC, Larsson A, Mans son B, Klareskog L, etal. Serum concentrations of cartilage oligomeric matrix protein, fibrinogen and hyaiuronan distinguish inflammation and cartilage destruction in experimental arthritis in rats. Rheumatology(Oxford), 2002:41(9):996-1000
[11]. Lu S, Carlsen S, Hansson AS, Holmdahl R. Immunization of rats with homologous type XI collagen leads to chronic and relapsing arthritis with different genetics and joint pathology than arthritis induced with homologous type Ⅱ collagen. J Autoimmun,2002;18(3):199-211
[12]. Romas E, Sims NA, Hards DK, Lindsay M, Quinn JW, Ryan PF, etal. Osteoprotegerin reduces osteoelast numbers and prevents bone erosion in Collagen-induecd arthritis. AmJPathol,2002:161(4):1419-27
[13].董文娟,梁秀军,翟泽玲,宋鸿儒等.穿山龙总皂苷对CIA大鼠滑膜血管新生的作用.[J].承德医学院学报,2010,27(4)：360-362
[14].于海荣,王济兴,陈建双等.穿山龙总皂苷含药血清对小鼠脾淋巴细胞增殖及IL-6产生影响的实验研究.[J].江苏中医药,2007,39(1)：57-58[15]. Remmers EF, Joe B, Griffiths MM, Dobbins DE, Dracheva SV, Hashirajnoto A, etal. Modulation of multiple experimental arthritis models by collagen-induced arthritis quantitative trait loci isolated in congenic rat lines:different effeets of non-major histocompatibility complex quantitative trait loci in males and females. Arthritis Rheum,2002; 46(8):2225-34
[16]. Bendele A, MeAbee T, Sennello G, Frazier J, ChliPala E, MeCabe D. Effieacy of sustained blood levels of interleukin-1 receptor antagonist in animal models of arthritis:comparison of efficacy in animal models with human clinical data. Arthritis Rheum,1999:42(3):498-506
[17]. Cuzzocrea S, MeDonald MC, Mota-FiliPe H, Mazzon E, Costantlno G, Britti D, etal. Beneficial effects of tempol, a membrane-permeable radical scavenger, in a rodent model of collagen-induced arthritis.Arthritis Rheum,2000:43(2):320-8
[18]. Mizuno M, Nishikawa K, Spiller OB, Morgan BP, Okada N, Okada H, etal. Membrane complement regulators Protect against the development of type II Collagen-induced arthritis in rats. Arthritis Rheum,2001: 44(10):2425-34
[19]. CatehPole B, Staines NA, Hamblin AS. Antigen Presentation of Type II collagen in rats. Clin ExP Immnunol,2001:125(3):478-84
[20]. Fjuisavva H. Nishimura T, Motohaga M. Effect of actarton type Ⅱ Collagen induced arthritis in mice. Arznein Forsh/Drug Res,1994:44(1):64-8
[21]. Lu S, Carlsen S, Hansson AS, Holmdahl R. Immunization of rats with homologous type Ⅺ collagen leads to chronic and relapsing arthritis with different genetics and joint Pathology than arthritis induced with homologous type Ⅱ collagen. J Autoimmun,2002;18(3):199-211
[22]. Han Z, Chang L, yamanishi Y, Karin M, Firestein GS. Joint damage and Inflammation in c-Jun N-tenminal kinase 2 knockout mice with passive murine Collagen-induced arthritis. Arthritis Rheum,2002:46(3):818-23
[23].魏伟,沈玉先,徐叔云.免疫细胞培养技术.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1421-22
[24].魏伟,沈玉先,徐叔云.淋巴细胞功能的测定.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1426-28
[25].沈玉先,魏伟,徐叔云.细胞因子的测定.见：徐叔云,卞如镰,陈修主编药理实验方法学(第三版).北京：人民卫生出版社,2002：1438-42
[26]. Cusehieri J, Gourlay D, Gareial I, Jelaeie S, Maier RV. Hypertonic preconditioning inhibits macrophage responsiveness to endotoxin. J Immunol,2002:168 (3):1389-96
[27]. Kiemer AK, Muller C, Vollmar AM. Inhibition of LPS-induced nitric oxide and TNF-alpha production by alpha-liPoic acid in rat KuPffer eells and in RAW 264.7 murine macrophages. Immunol Cell Biol,2002:80(6):550-7
[28].Sugiyama T, Ishii S, Yamamoto J, Irie R, Saito K, Otuki T, etal. cDNA macroarray analysis of gene expression in synoviocytes stimulated with TNF alpha. FEBS Lett,2002;517(1-3):121-8
[29].Privat J, Oo A, Waterbury LD. Production of interleukin-6, but not interleukin-8, induced by TNF-alpha or IL-1 beta in human f ibroblast-like ynoviocyte in ercreases over cell passage. Proc West Pharmacol Soc, 2001:44:9-13
[30]. Migita K, Thnaka F,s Yamasakis, Shibatomi K, Ida H, Kawakami A, etal. Regulation of rheumatoid synoviocyte Proliferation by endogenous P53 induction. Clin ExP Immunol,2001:126(2):334-8
[31].Nakazawa M, Ishii H, Aono H, Takai M, Honda T, Aiatanl S, etal. Role of Noteh-1 intral cellular domain in activation of rheumatoid synovioeytes.ArthritisRheum,2001:44(7):1545-54
[32]. Trentham DE. Townes AS, Kang AH. Autoimmunity to type Ⅱ collagen:an Experimental model of arthritis. J Exp Med,1977:146:857-66
[33], Romas E, Bakharevski O, Hards DK, Kartsogiannis V, Quinn JM, Ryan PF, etal.Expression of osteoelast differentiation factor at sites of bone erosion in collagen-induced arthritis. Arthritis Rheum,2000;43(4):821-6
[34]. Kobayashi K, Nakashima A, Nagata H, Nakajima H,、Yamaguehi K, Sato S, etal. Anti-arthritic effects of KF20444, a new immunosuppressive compound inhibiting Dihydroorotate dehydrogenase, on rat collagen-inducedarthritis.Inflamm Res, 2001:50(1):24-31
[35]. Nawata K, EnokidaM,、Yamasaki D, Minamizaki T, Hagino H, Morio Y etal. Tensile Properties of rat anterior cruciate ligament in collagen induced arthritis. Ann Rheum Dis,2001:60(4):395-8
[36].Hoshino K, Hanyu T, Arai K, Takahashi HE.Mineraldensity and histomorphometric assessment of bone changes in the proximal tibia early after induction of type Ⅱ Collagen-induced arthritis in growing and mature rats. J Bone Miner Metab,2001;19(2):76-83
[37]. Cuzzoerea S, Mazzon E, Bevilaqua C, Costantlno G, Britti D, Mazzullo G, etal. Clorieromene, a coumarine derivative, protects against collagen-induced arthritis in Lewis rats. Br J phannacol,2000:131 (7):1399-407
[38]. Yamasaki D, Enokida M, Okano T, Hagino H, Teshima R. Effects of ovariectomy and estrogen replacement therapy on arthritis and bone mineral density in rats with collagen-induced arthritis. Bone,2001:28(6):634-40
[39]. Wernhoff P, Unger C, Bajiner E, Burkhardt H, Holmdahl R. Identification of conformnation-dependent epitopes and V gene selection in the B cell response to type Ⅱ collagen in the DA rat. Int Immunol,2001; 13(7):909-19
[40]. CatehPole B, Staines NA, Hamblin AS. Antigen Presentation of Type Ⅱ collagen in rats.Clin Exp Immunol,2001:125(3):478-84
[41]. Romas E, Bakharevski O, Hards DK, Kartsogiarmis V, Quinn JM, Ryan PF, etal. Expression of osteoelast differentiation factor at sites of bone erosion in collagen-induced arthritis. Arthritis Rheum,2000;43 (4):821-6
[42]. Cheon, Yu SJ, Yoo DH, Chae IJ, Song GG, Sohn J. Increased expression of Pro-inflammatory cytokines and metal loproteinase-1 by TGF-beta 1 in synovial fibroblasts from rheumatoid arthritis and normal individuals. Clin ExP Immunol,2002:127(3):547-52
[43]. Brinkman DM, ten Cate R,Vossen JM.Smeeet TJ,Kraan MC,Tak PP. Decrease in syovial cellularity and cytokine expression after autologous stem cell transplantation in patients with juvenile idiopathic arthritis. ArthritisRheum,2002:46(4):1121-3
[44]. Okazaki A, Koshino T, Saito T, Takagi T. Osseous tissue reaction around Hydroxyapatite block implanted into proximal metaphysic of tibia of rat with collagen-induced arthritis. Biomaterials,2000:21(5):483-7
[45]. He W, Pelletier JP, Martel-Pelletier J, Laufer S, Di Battista JA.Synthesis of interleukinlbeta, tumornecrosis factor-alpha.and interstitial collagenase(MMP-1)15 eicosanoid dependent in human osteo arthritis synovial membrane explants:inieractions with anti-inflammatory cytokines.J Rheumatol,2002;29(3):546-53
[46].Matsuura M, Imayoshi T, Okumoto T. Effect of FTY720, a novel immunosuppressant, on adjuvant-and collagen-induced arthritis in rats. Int J Immunopharmacol,2000:22(4):323-31.
[47].Du Z F,Li Z B,Sun C X,et al. Effects of Artesunate on TNF-a and IL-1 in the serum of rats with ajuvant arthritis[J].Guangdong Medical Journal,2010,31 (13):1637-40.
[48]. Xie S J,Song H R.The Experimental Study on Immunoregulatory Effects of Whole Saponin of Rhizoma D ioscreae Nipponicae on AA Rat[J]. LiaoNing Journal of Traditional Chinese Medicine,2007,34 (9):1323-1325
[49].Li Y,Song Y Y,Zhang H Q.Effect of Echinacoside on immune function and mitochondrial DNA relative content of aging mice[J]. Chin Pharmaco Bull,2010,26 (6):810-3.
[50].Yun CX,Yu XL,Wu G et al. Effect of Sangyehuang mixture on cellular immunological function in rats[J]. Lishizhen Medicine Materia Medica Research,2009,20(5):1155-7.
[51]. Trentham DE.Townes AS, Kang AH. Autoimmunity to type Ⅱ collagen:an experimental model of arthritis.J Exp Med,1977;146:857-66
[52].Nawata K,Enokida M,Yamasaki D,Minamizaki T,Hagino H,Morio Y, et al. Tensile properties of rat anterior cruciate ligament in collagen induced arthritis. Ann Rheum Dis,2001;60(4):395-8
[53].Cai H, Zheng Z L,Shang W,et al.The effect of curcumin on the immune organ weight and pathomorphology of adjuvant arthritis rats[J].JETCM,2009,18(4):594-5.
[54].Yang X H,Wu J,Guo L J. Optimization on extraction technology of saponin of chaenomeles speciosa and study on its anti-inflammatory immunologic mechanism to rat adjuvant arthritis[J].2010,25(5):547-9.
[55].Yi J F,Lu A P. Study on the Immunology of Triptolide and Dihydroartem-isinine Compound Treating CIA Rats[J].Journal of Jiangxi university of TCM,2008,20(6):49-51.
[56].GAO YX,LIANG XJ,DONG WJ,et al. Effects of Total Saponin from Rhizoma Dioscreae Nipponicae on NF-κB p65 Activity and STAT3 Protein Expression in Joint Synovium of CIA Rats[J]. Journal of China Medical University Vol.41 No.6 Jun.2012, 41(6):485-489
[57].WANG XP, LU X. New progress of Dioscin biological activity research [J]. International Journal of Traditional Chinese Medicine,2004,26(3):138-141.
[58].LI SP, HU XM,JIN ZM. Therapeutic Effect of Dioscin on rheumatoid arthritis [J]. Zhejiang clinical medicine,2008,11(10):628.
[59].YANG JC, CHEN ZX. Medical Molecular Biology.BeiJing:Chemical Industry Press,2003:1204.
[60].YAO L, LIU SM. Study on the Effective Ingredients of Rhizoma Dioscoreae Nipponicae on Experimental Acute Gouty Arthritis[J]. Chinese Archives of Traditional Chinese Medicine,2010,28(8):1724-1726
[61].Yang X H,Wu J,Guo L J. Optimization on extraction technology of saponin of chaenomeles speciosa and study on its anti-inflammatory immunologic mechanism to rat adjuvant arthritis[J].2010,25(5):547-9.
[62].Yi J F, Lu A P. Study on the Immunology of Triptolide and Dihydroartem-isinine Compound Treating CIA Rats[J] Journal of Jiangxi university of TCM,2008,20(6):49-51.
[63].GM.Campo, A.Avenoso, S.Campo, et al. Chondroitin-4-sulphate inhibitsNF-κB translocation and caspase activation in collagen-induced arthritis inmice. Osteoarthritis and Cartilage,2008,4:1.
[64].DING Q, YOU ZL. Effects of compound components from Panax Notoginseng on NF-κBP65 and TNF-a, IL-β in inflammation model of human endometrial cell. [J] Chinese Journal of Traditional Medical Science and Technology,2007,14(2):113.
[65].Anderson GD,KeysKL,De Ciechi PA, et al Combination therapies that inhibit cyclooxygenase-2 and leukotriene synthesis prevent disease inmurine collagen induced arthritis. Inflamm Res,2009,2:109.
[66].Noguchi M, Kimoto A, Sasamata M, et al Micro-CT imaging analysis for the effect of celecoxib, a cyclooxygenase-2 inhibitor, on inflammatory bone destruction in adjuvant arthritis rats. J Bone Miner Metab,2008,26:461.
[67].Lee HS, Lee CH, Tsai HC, et al Inhibition of cyclooxygenase 2 expression by diallyl sulfide on joint inflammation induced by urate crystal and IL-1β.OsteoarthritisCartilage,2009,1:91.
[68].Gabay C.Cytokine inhibitors in the treatment of rheumatoid arthritis. Expert OPin Biol Ther,2002:2(2):135-49
[69]. Schrnitz M L, Baeuerle P A. The p65 subunnit is responsible for the strong transcription activating potential of NF-κB. EMBO J,1991; 10(12):3805
[70]. MLienhard S, MarcosA, Patrick A. Transactivation domain 2 (TA2) of p65 NF-κB. J. Biol. Chem,1995; 270(26):15576
[71]. Llui's E, Julia IE, Alex RM, et al. I-κB and p65 regulate the cyto-plasmic shuttling of ruclear corepressors:Cross-talk between notch and NF-κB pathways. Molecular Biology of the Cells,2003;14:491
[72].宋震坤,陈文照.环氧化酶-2与骨关节炎[J].中国骨伤,2003,13(5)：314-315.
[73].吴尚洁,蔡颖,赵水平.阿托伐他汀降低脂多糖诱导下人肺上皮细胞COX-2的表达[J].中国医师杂志,2004,6(6)：768.
[74].邓渝林,陶霞,周静等.第二代COX-2抑制剂的研究进展[J].解放军药学学报,2005,21(3)：209.
[75].戴生明,韩星海,施冶青.两种选择性COX-2抑制剂治疗类风湿关节炎和骨关节炎的临床比较[J].中华风湿病学杂志,2000,4(6)：378.
[76].唐福林,张奉春,郑文洁等.双醋瑞因治疗膝骨关节炎的疗效与安全性多中心对照研究[J].中华风湿病学杂志,2005,9(7)：423.
[77].皮治兵,蒋宗滨.COX-2抑制剂临床研究新进展[J].中国疼痛医学杂志,2006,12(5)：297.
[1]van Boekel MA, Erik RV, van den Hoogen FH, et al. Autoantibody systems in rheumatoid arthritis:specificity, sensitivity and diagnostic value. Arthritis Res,2002,4:87-93.
[2]Egeland T, Munthe E. The role of the laboratory in rheumatology. Rheumatoid factors. Clin Rheum Dis 1983,9(1):135-60.
[3]MagsaamJ, Ferjenck P, Tenmpels M, et al. A new method for the detection of hidden IgM rheumatoid factor in patients with juvenile rheumatoid arthritis[J].J Rheumatol 1987,14(5):964-967.
[4]何菁,贾汝琳,韩蕾,等.隐性类风湿因子在类风湿关节炎诊断中的意义[J].中华风湿病杂志,2001,5：24-27·
[5]Hoet RM, van Venrooij WJ. The antiperinuclear factor and antikeratin antibodies in rheumatoid arthritis. In:Smolen JS, Kalden JR, Maini RN, editors. Rheumatoid arthritis. Berlin: Springer Verlag; 1992. p.299-318.
[6]]Young BJ, Mallya RK, Leslie RD, Clark CJ, Hamblin TJ. Antikeratin antibodies in rheumatoid arthritis. Br Med J 1979; 2:97-9.
[7]穆荣,孙晓云,栗占国.类风湿因子和抗胍氨酸多肽抗体联合检测在类风湿关节炎诊断中的意义[J].北京大学学报,2005,37(5)：498-500.
[8]Forslind K, AhlmenM, EberhardtK, et al. Prediction of radiological outcome in early rheumatoid arthritis in clinical practice:role of antibodies to citrullinated peptides (anti-CCP) [J]. Ann Rheum Dis,2004, 63(9):1090-1095.
[9]El-Gabalawy HS, Wilkins JA. Anti-Sa antibodies:prognostic and pathogenetic significance to rheumatoid arthritis. Arthritis Res Ther, 2004,6:86-9.
[10]Hayem G, Chazerain P, Combe B, et al. Anti-Sa antibody is an accurate diagnostic and prognostic marker in adult rheumatoid arthritis [J]. J Rheumatol,1999;26(1):7-13
[11]Goldbach-Mansky R, Lee J, McCoy A, et al. Rheumatoid arthritis associated autoantibodies in patients with synovitis of recent onset. Arthritis Res,2000,2:236-243.
[12]钱龙,曾庆馀,黄少弼,等.抗核周因子的检测及其在类风湿关节炎中的意义[J].中华风湿病学杂志,1999;3(2)：100-103
[13]Schellekens GA, Visser H, de Jong BAW, et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide[J]. Artritis Rheum,2000; 43(1):155-163
[14]Blass S, Union A, Raymackers J, et al. The stress protein BiP is overexpressed and is a major B and T cell target in rheumatoid arthritis. Arthritis Rheum,2001,44:761-771.
[15]田昕,毕黎琦,栗占国.抗瓜氨酸化Ⅱ型胶原抗体的检测及其与类风湿关节炎的相关性研究[J].中华医学杂志,2006,86(33)：2334.
[16]Burkhardt H, Sehnert B, Bockermann R, et al. Humoral immune response to citrullinated collagen type Ⅱ determinants in early rheumatoid arthritis [J].Eur J Imm uno,12005,35(5):1643-1652.
[17]Nielen MM, vander Horst AR, van Sehaardenbug D, et al. Antibodies to citrullinated human fibrinogen(ACF) have diagnostic and prognostic value in early arthritis. Ann Rheum Dis,2005,64:1199-1204.
[18]赵义,田听,栗占国.抗瓜氨酸化纤维蛋白原抗体的检测及其在类风湿关节炎中的临床意义.北京大学学报,2006,38：350.
[19]Bang H, Luthke K, Burmester GR, et al. Mutated citrullinated Vimentin as a candidate autoantigen for diagnosis and monitoring of Disease activity in Rheumatoid Arthritis. Ann Rheum Dis,2006,65:144.
[20]姜东林,孙钧铭,姜升阳,等.类风湿关节炎患者抗MCV、抗CCP抗体与RF诊断价值比较[J].临床检验杂志,2009,27(2)：137-138.
[1]van Boekel MA, Erik RV, van den Hoogen FH, et al. Autoantibody systems in rheumatoid arthritis:specificity, sensitivity and diagnostic value. Arthritis Res, 2002,4:87-93.
[2]何菁,贾汝琳,韩蕾,等.隐性类风湿因子在类风湿关节炎诊断中的意义[J].中华风湿病杂志,2001,5：24-27.
[3]穆荣,孙晓云,栗占国.类风湿因子和抗胍氨酸多肽抗体联合检测在类风湿关节炎诊断中的意义[J].北京大学学报,2005,37(5)：498-500.
[4]Forslind K, Ahlmen M, Eberhardt K, et al. Prediction of radiological outcome in early rheumatoid arthritis in clinical practice:role of antibodies to citrullinated peptides (anti-CCP) [J].Ann Rheum Dis,2004,63(9):1090-1095.
[5]E1-Gabalawy HS, Wilkins JA. Anti-Sa antibodies:prognostic and pathogenetic significance to rheumatoid arthritis. Arthritis Res Ther,2004,6:86-9.
[6]Goldbach-Mansky R, Lee J, McCoy A, et al. Rheumatoid arthritis associated autoantibodies in patients with synovitis of recent onset. Arthritis Res, 2000,2:236-243.
[7]钱龙,曾庆馀,黄少弼,等.抗核周因子的检测及其在类风湿关节炎中的意义[J].中华风湿病学杂志,1999：3(2)：100-103
[8]Schellekens GA, Visser H, de Jong BAW, et al. The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide[J]. Artritis Rheum,2000; 43(1):155-163
[9]Blass S, Union A, Raymackers J, et al. The stress protein BiP is overexpressed and is a major B and T cell target in rheumatoid arthritis. Arthritis Rheum, 2001,44:761-771.
[10]田昕,毕黎琦,栗占国.抗瓜氨酸化Ⅱ型胶原抗体的检测及其与类风湿关节炎的相关性研究[J].中华医学杂志,2006,86(33)：2334.
[11]Burkhardt H, Sehnert B, Bockermann R, et al. Humoral immune response to citrullinated collagen typeⅡdeterminants in early rheumatoid arthritis [J].Eur J Immunol 2005,35(5):1643-1652.
[12]Nielen MM, vander Horst AR, van Sehaardenbug D, et al. Antibodies to citrullinated human fibrinogen(ACF) have diagnostic and prognostic value in early arthritis. Ann Rheum Dis,2005,64:1199-1204.
[13]赵义,田听,栗占国.抗瓜氨酸化纤维蛋白原抗体的检测及其在类风湿关节炎中的临床意义.北京大学学报,2006,38：350.
[14]Bang H, Luthke K, Burmester GR, et al. Mutated citrullinatedVimentin as a candidate autoantigen for diagnosis and monitoring of Disease activity in Rheumatoid Arthritis. Ann Rheum Dis,2006,65:144.
[15]姜东林,孙钧铭,姜升阳,等.类风湿关节炎患者抗MCV、抗CCP抗体与RF诊断价值比较[J].临床检验杂志,2009,27(2)：137-138.