竹节参皂苷抗炎抗风湿作用及机理研究
|
摘要
类风湿性关节炎(RA)是一种慢性、炎性、系统性的自身免疫性疾病,以关节和关节周围组织非化脓性炎症为主要特征,成为继缺血性心脏病后导致工作能力丧失的第二位疾病,因其高度致残率WHO将其列为重点关注和治疗的疾病之一。传统的治疗RA的药物大多作用途径或靶点单一,同时有一定的毒副作用,因此开发高效、低毒、多靶点作用的新药就显得尤为重要和迫切。
竹节参(Panax japonicum C.A.Mey.)五加科植物,中医记载其具有滋补强壮、散瘀止痛、止血、祛痰功能。用于病后虚弱、肺结核咯血、咳嗽痰多、跌打损伤。本实验将系统开展其抗炎抗风湿作用及机理研究。
1) TSPJ对小鼠常规的炎症反应的影响：
(1) TSPJ对角叉菜胶所致小鼠足肿胀具有抑制作用,TSPJ中高剂量均能明显减轻角叉菜胶所致小鼠右后足的红、肿等症状,50 mg·kg-1 TSPJ组在致炎后1-3 h,100 mg·kg-1 TSPJ组在致炎后0.5-4 h减轻足肿胀,与control组相比,有明显差异(P<0.05,P<0.01)。
(2)TSPJ对巴豆油所致小鼠耳肿胀具有抑制作用,TSPJ高中组均能明显减轻巴豆油所致小鼠右耳的红、肿等症状,显著抑制巴豆油所致的小鼠耳肿胀,100mg·k-1 TSPJ组的抑制率可达33.33%,作用强度与剂量呈正相关,与control组比较,差异有显著性(P<0.05,P<0.01)。
(3)TSPJ对冰醋酸所致小鼠腹腔毛细血管通透性增高有抑制作用,各组均能明显抑制冰醋酸所致小鼠腹腔毛细血管通透性的增高,100 mg·kg-1 TSPJ组的抑制率可达52.02%,作用强度与剂量呈正相关,与control组比较,差异有显著性(P<0.05,P<0.01)。
(4) TSPJ具有提高血清中GSH-PX、SOD活性及降低MDA的作用,TSPJ各组血清GSH-PX, SOD活性增高,血清MDA含量下降,与control组比较,差异具有显著性(P<0.05)。
(5) TSPJ使炎症小鼠血清中IL-1, TNF-α含量下降,TSPJ中高剂量组小鼠血清IL-1水平下降(P<0.05), TSPJ高剂量组小鼠血清TNF-α水平下降,差异性显著(P<0.01)。
2) TSPJ对CIA大鼠组织病理学及超微结构改变的影响
(1) TSPJ各剂量组AI评分也随着继发性炎症的产生而升高,但与同期的模型组比较有显著性差异,提示竹节参总皂苷具有抗CIA的作用,且具有量效关系,但治疗效果比雷公藤治疗组稍差。
(2)模型组大鼠致炎后d12起的足爪开始肿胀,d24-d28达到高峰,至d36仍有一定程度的肿胀。TSPJ各剂量组肿胀程度也随着继发性炎症的产生而增加,但与同期的模型组比较有显著性差异,提示TSPJ具有抗CIA的作用,且具有量效关系,但治疗效果比雷公藤治疗组稍差。
(3)CIA模型组进入继发性炎症阶段(d18以后)体重增长速度减慢或停止,TPT治疗组体重增长速度也出现减慢现象,这可能是TPT在治疗关节炎的同时产生了一定的胃肠道刺激导致大鼠进食量下降所致,TSPJ各组体重增长速度基本正常。
(4)CIA模型可导致机体免疫功能亢进；给药组大鼠的脾脏指数、胸腺指数与模型组比较均有下降,其中TPT组差异具有显著性(p<0.05),其余组有下降趋势但无明显差异,表明TSPJ具有使亢进的免疫功能恢复正常的作用。
(5) TSPJ具有改善CIA关节病变作用：TSPJ各剂量组均能不同程度的减轻关节软组织的充血水肿,足趾容积及关节评分均低于模型组；减轻滑膜充血、水肿的程度；明显抑制CIA大鼠滑膜增生；抑制炎性细胞浸润；抑制大鼠软骨细胞增生。
(6) ConA诱导的CIA大鼠脾脏T淋巴细胞增殖反应明显低于正常对照组(P<0.01),产生的IL-2水平明显降低,TSPJ给药可以恢复CIA大鼠脾淋巴细胞功能,TPT对CIA大鼠低下的脾淋巴细胞增殖反应及产生IL-2没有明显影响。
(7)CIA组PMΦIL-1β、IL-6和TNF-α等致炎细胞因子的表达量均明显高于正常组(P<0.01),表明在CIA模型产生过程中,各种致炎因子大量产生参与炎症反应。TSPJ大中剂量能明显抑制PMφ产生致炎细胞因子IL-1β、IL-6和TNF-α水平。
(8)CIA滑膜超微结构显示：滑膜层细胞内膜明显增厚,多达5-6层,滑膜细胞增生肥大,排列紊乱。细胞器变形,甚至消失,胞质内充满大量空泡,胞核染色质分布不均。有的细胞因变性、坏死至萎缩,仅见到细胞碎块与大量胶原纤维交错排列。TSPJ可明显改善CIA滑膜超微结构,滑膜层细胞性内膜无增厚,细胞排列均匀,偶见少量淋巴细胞浸润,巨噬细胞型细胞以初级溶酶体多见、线粒体稍增多,成纤维细胞型细胞的胞浆内富含粗面内质网及核糖小体,各型细胞接近正常。
(9)CIA模型组关节滑膜在炎症过程中,ICAM-1产量(阳性细胞总数、平均黑度值、面积总和、积分光密度值)明显增加,TSPJ各剂量组关节滑膜ICAM-1表达数目、面积总和、积分光密度值及平均黑度值均低于模型对照组,且有着性差异(P<0.05或P<0.01)。
(10)正常以及CIA滑膜细胞培养和细胞因子的研究表明：正常滑膜细胞在未受到刺激时并无细胞因子如IL-1、IL-6、IL-8和TNF-α分泌,细胞增长速度正常；当正常滑膜细胞接受LPS刺激后,细胞因子如IL-1、IL-6、IL-8和TNF-α分泌量增加,细胞增殖速度加快,表明LPS是一种良好微生物抗原刺激剂,可以诱导正常细胞的增殖分化。CIA滑膜细胞未受刺激也会有少量细胞因子分泌,当受到LPS刺激后,细胞增殖速度明显加快,细胞因子分泌量成倍或数十倍增加,表明CIA滑膜细胞本身已经处于一种敏感状态,对外来抗原刺激极为敏感,诱发炎症反应的恶性循环。雷公藤显示出强大的抗炎作用；TSPJ竹节参在滑膜细胞培养试验中未观察到细胞损伤现象,并无刺激细胞因子分泌作用,仅使处于“高敏”的CIA滑膜细胞状态恢复正常。
从本实验结果来看,竹节参具有以下作用：(1)提高机体抗氧化能力,减少自由基对炎症组织的损伤；(2)具有减轻炎症组织中炎症介质PGE和LTB的作用；(3)使炎症组织中细胞因子如IL-1B、IL-6、TNF-α等明显减少；(4)减轻炎症组织中ICAM-1的产生,使炎症部位不产生粘连；(5)对免疫系统具有“双向调节作用”,在治疗过程中使亢进的免疫功能下调至正常,不会产生类似雷公藤和其他免疫抑制剂导致的免疫功能减退。因此,竹节参极有可能开发成为一种有效、无毒或低毒的抗炎抗风湿的新药。
Rheumatoid arthritis (RA) is a chronic, inflammatory and systemic autoimmune disease, which joints and joint's surrounding tissue non-suppurative inflammations are its main features. It becomes the second cause of loss ability to work behind the ischemic heart disease. The traditional treatment of RA drugs are mostly single pathway or target, while there are certain side effects, so development of efficient, low toxicity, the role of multi-target drug is particularly important and urgent.
Panax japonicus (Panax japonicum CA Mey.),a traditional chinese medical herb, belongs to Araliaceae. It has strong tonic, dissipate blood stasis and alleviate pain, bleeding and expectorant functions in Chinese medicine record.So it used totreated weakness after diseases, tuberculosis hemoptysis, cough, sputum, and bruises. In this study, its anti-inflammatory effects and anti-rheumatic mechanisms will be carried out syetemic researched.
1) The effect of TSPJ on normal inflammatory response in mice:
(1) TSPJ 50 mg/kg and 100 mg/kg group could significantly reduce the paw edema, foot redness swelling and other symptoms induced by carrageenan after inflammation 0.5～4 h (P<0.05)Vs control group.
(2) TSPJ could inhibit the mouse ear edema induced by croton oil, the ear's redness, swelling and other symptoms are significantly inhibited.The high dose group of TSPJ's inhibition rate is 33.33% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).
(3) TSPJ could inhibit the increased capillary permeability induced by acetic acid on mice inhibition rate of TSPJ's high dose group(100 mg/kg) is 48.6% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).
(4) TSPJ could increasethe levels of serum GSH-PX, SOD activity and reduce serum MDA content. The inhibition rate is positively correlated with the dose
(5) TSPJ could reduce the inflammatory factors (IL-1, TNF-α) in mice serum, the differences are significant (P<0.05).
2) The effect of TSPJ on CIA rats'histopathological and ultrastructural stractures
(1) The AI scores in CIA group are higher than normal group, three dose group's AI of TSPJ were also increased compare with the normal group at the same period, but the difference were significant compare with the CIA group at the same period.
(2) The model group' feet swelling begins from d12 to the end, peak time was at d24-d28. The swelling degree of TSPJ groups were also increased compare with the normal group at the same period, but the difference was significant compare with the CIA group at the same period.
(3) CIA model rats'weight-gain rate slow down or stoped at the secondary inflammatory phase (after d18), TPT treatment group also shown the weight-gain rate, TSPJ groups' growth rate were normal.
(4) The immune function of CIA model could lead to higher compared with model group; the administration of TPT could decreasethe higher s spleen index and thymus index in CIA rats, the difference was significant,and TSPJ group alos decline differences but no significant.
(5) TSPJ can improve joint damage of CIA:thre dose groups of TSPJ can reduce the levels of congestion and edema of soft tissue joint, toe joint volume and scores; reduce synovial hyperemia, edema of the degree; significantly inhibit the CIA rats' synovial hyperplasia; inhibit the inflammatory cell' infiltration; inhibit chondrocytes proliferation.
(6) ConA-induced spleen T lymphocyte proliferation in CIA group was significantly lower than the control group, and IL-2 levels were also significantly lower, TSPJ groups can restore the CIA rat spleen lymphocytes and production of IL-2, but TPT group has no effect on proliferation of lymphocytes and production of IL-2.
(7) The productions of IL-1β, IL-6 and TNF-αwere significantly higher in CIA group's PMΦthan those of normal group (P<0.05), TSPJ medium and large doses groups can inhibit the production of proinflammatory cytokines IL-1β, IL-6 and TNF-αlevels in CIA's PMΦ.
(8) The synovial membrane ultrastructure:at the CIA group, the synovial cell lining layers were thickened up to 5-6 layers and disarrangement, organelle deformation, or even disappear, the cytoplasm filled with large vacuoles, the uneven distribution of nuclear chromatin. Some cells due to degeneration and necrosis to atrophy, only to see a large number of cell fragments with staggered collagen fibers. TSPJ can significantly improve the CIA synovial ultrastructure of synovial lining cell layer and cell arrangement, only occasionally a small amount of lymphocytic infiltration can be seen, A-type cells to primary lysosomes, mitochondria slightly increased, B-type Cytoplasm of cells rich in rough endoplasmic reticulum and ribosome bodies, various types of cells close to normal.
(9) The ICAM-1 production (the total number of positive cells, the average black level value of the total area and integral optical density) were significantly increased in CIA model group's synovial membrane during the inflammatory process, TSPJ groups can reducethe ICAM-1 expression in synovial number, size Sum, integral optical density and average black levels, the differences are significant (P<0.05).
(10) synovial cell culture and cytokine researches showed:normal synovial cells' cytokines productions are very low even no detected, cell growth rate are normal; after stimulated by LPS, the speed of cell proliferation and cytokines such as IL-1, IL-6, IL-8 and TNF-αsecretions are increased, this suggest that LPS is a good stimulant of microbial antigens, can induce normal cell proliferation and differentiation. CIA synovial cells' proliferation rate and cytokines secretions were significantly higher after LPS stimulation. Tripterygium showed a strong anti-inflammatory effects; TSPJ didn't show cell-damage in the synovial cell culture experiments,and didn't stimulate cytokine secretion, only regulate the CIA synovial cells from "super- sensitivity" status to normal.
From the experimental results, TSPJ has the following parameters:(1) increasing the antioxidant capacity, reduce inflammation, tissue damage induced by free radicals; (2) to relieve inflammatory mediators such as PGE and LTB in inflamed tissue; (3) significantly decreased cell factors such as IL-1B, IL-6, TNF-αinf lammated tissues; (4) reduce the ICAM-1 generation in inflammated tissues; (5) TSPJ has 'two-way adjustment' on the immune system, so at the process of treating RA it can reduce hyperthyroidism immune function to normal, but don't produce immune dysfunction as triptolide and other immunosuppressive agents do. Therefore, TSPJ is likely to be developed as an effective, non-toxic or low toxic anti-inflammatory and anti- rheumatic drug.
竹节参(Panax japonicum C.A.Mey.)五加科植物,中医记载其具有滋补强壮、散瘀止痛、止血、祛痰功能。用于病后虚弱、肺结核咯血、咳嗽痰多、跌打损伤。本实验将系统开展其抗炎抗风湿作用及机理研究。
1) TSPJ对小鼠常规的炎症反应的影响：
(1) TSPJ对角叉菜胶所致小鼠足肿胀具有抑制作用,TSPJ中高剂量均能明显减轻角叉菜胶所致小鼠右后足的红、肿等症状,50 mg·kg-1 TSPJ组在致炎后1-3 h,100 mg·kg-1 TSPJ组在致炎后0.5-4 h减轻足肿胀,与control组相比,有明显差异(P<0.05,P<0.01)。
(2)TSPJ对巴豆油所致小鼠耳肿胀具有抑制作用,TSPJ高中组均能明显减轻巴豆油所致小鼠右耳的红、肿等症状,显著抑制巴豆油所致的小鼠耳肿胀,100mg·k-1 TSPJ组的抑制率可达33.33%,作用强度与剂量呈正相关,与control组比较,差异有显著性(P<0.05,P<0.01)。
(3)TSPJ对冰醋酸所致小鼠腹腔毛细血管通透性增高有抑制作用,各组均能明显抑制冰醋酸所致小鼠腹腔毛细血管通透性的增高,100 mg·kg-1 TSPJ组的抑制率可达52.02%,作用强度与剂量呈正相关,与control组比较,差异有显著性(P<0.05,P<0.01)。
(4) TSPJ具有提高血清中GSH-PX、SOD活性及降低MDA的作用,TSPJ各组血清GSH-PX, SOD活性增高,血清MDA含量下降,与control组比较,差异具有显著性(P<0.05)。
(5) TSPJ使炎症小鼠血清中IL-1, TNF-α含量下降,TSPJ中高剂量组小鼠血清IL-1水平下降(P<0.05), TSPJ高剂量组小鼠血清TNF-α水平下降,差异性显著(P<0.01)。
2) TSPJ对CIA大鼠组织病理学及超微结构改变的影响
(1) TSPJ各剂量组AI评分也随着继发性炎症的产生而升高,但与同期的模型组比较有显著性差异,提示竹节参总皂苷具有抗CIA的作用,且具有量效关系,但治疗效果比雷公藤治疗组稍差。
(2)模型组大鼠致炎后d12起的足爪开始肿胀,d24-d28达到高峰,至d36仍有一定程度的肿胀。TSPJ各剂量组肿胀程度也随着继发性炎症的产生而增加,但与同期的模型组比较有显著性差异,提示TSPJ具有抗CIA的作用,且具有量效关系,但治疗效果比雷公藤治疗组稍差。
(3)CIA模型组进入继发性炎症阶段(d18以后)体重增长速度减慢或停止,TPT治疗组体重增长速度也出现减慢现象,这可能是TPT在治疗关节炎的同时产生了一定的胃肠道刺激导致大鼠进食量下降所致,TSPJ各组体重增长速度基本正常。
(4)CIA模型可导致机体免疫功能亢进；给药组大鼠的脾脏指数、胸腺指数与模型组比较均有下降,其中TPT组差异具有显著性(p<0.05),其余组有下降趋势但无明显差异,表明TSPJ具有使亢进的免疫功能恢复正常的作用。
(5) TSPJ具有改善CIA关节病变作用：TSPJ各剂量组均能不同程度的减轻关节软组织的充血水肿,足趾容积及关节评分均低于模型组；减轻滑膜充血、水肿的程度；明显抑制CIA大鼠滑膜增生；抑制炎性细胞浸润；抑制大鼠软骨细胞增生。
(6) ConA诱导的CIA大鼠脾脏T淋巴细胞增殖反应明显低于正常对照组(P<0.01),产生的IL-2水平明显降低,TSPJ给药可以恢复CIA大鼠脾淋巴细胞功能,TPT对CIA大鼠低下的脾淋巴细胞增殖反应及产生IL-2没有明显影响。
(7)CIA组PMΦIL-1β、IL-6和TNF-α等致炎细胞因子的表达量均明显高于正常组(P<0.01),表明在CIA模型产生过程中,各种致炎因子大量产生参与炎症反应。TSPJ大中剂量能明显抑制PMφ产生致炎细胞因子IL-1β、IL-6和TNF-α水平。
(8)CIA滑膜超微结构显示：滑膜层细胞内膜明显增厚,多达5-6层,滑膜细胞增生肥大,排列紊乱。细胞器变形,甚至消失,胞质内充满大量空泡,胞核染色质分布不均。有的细胞因变性、坏死至萎缩,仅见到细胞碎块与大量胶原纤维交错排列。TSPJ可明显改善CIA滑膜超微结构,滑膜层细胞性内膜无增厚,细胞排列均匀,偶见少量淋巴细胞浸润,巨噬细胞型细胞以初级溶酶体多见、线粒体稍增多,成纤维细胞型细胞的胞浆内富含粗面内质网及核糖小体,各型细胞接近正常。
(9)CIA模型组关节滑膜在炎症过程中,ICAM-1产量(阳性细胞总数、平均黑度值、面积总和、积分光密度值)明显增加,TSPJ各剂量组关节滑膜ICAM-1表达数目、面积总和、积分光密度值及平均黑度值均低于模型对照组,且有着性差异(P<0.05或P<0.01)。
(10)正常以及CIA滑膜细胞培养和细胞因子的研究表明：正常滑膜细胞在未受到刺激时并无细胞因子如IL-1、IL-6、IL-8和TNF-α分泌,细胞增长速度正常；当正常滑膜细胞接受LPS刺激后,细胞因子如IL-1、IL-6、IL-8和TNF-α分泌量增加,细胞增殖速度加快,表明LPS是一种良好微生物抗原刺激剂,可以诱导正常细胞的增殖分化。CIA滑膜细胞未受刺激也会有少量细胞因子分泌,当受到LPS刺激后,细胞增殖速度明显加快,细胞因子分泌量成倍或数十倍增加,表明CIA滑膜细胞本身已经处于一种敏感状态,对外来抗原刺激极为敏感,诱发炎症反应的恶性循环。雷公藤显示出强大的抗炎作用；TSPJ竹节参在滑膜细胞培养试验中未观察到细胞损伤现象,并无刺激细胞因子分泌作用,仅使处于“高敏”的CIA滑膜细胞状态恢复正常。
从本实验结果来看,竹节参具有以下作用：(1)提高机体抗氧化能力,减少自由基对炎症组织的损伤；(2)具有减轻炎症组织中炎症介质PGE和LTB的作用；(3)使炎症组织中细胞因子如IL-1B、IL-6、TNF-α等明显减少；(4)减轻炎症组织中ICAM-1的产生,使炎症部位不产生粘连；(5)对免疫系统具有“双向调节作用”,在治疗过程中使亢进的免疫功能下调至正常,不会产生类似雷公藤和其他免疫抑制剂导致的免疫功能减退。因此,竹节参极有可能开发成为一种有效、无毒或低毒的抗炎抗风湿的新药。
Rheumatoid arthritis (RA) is a chronic, inflammatory and systemic autoimmune disease, which joints and joint's surrounding tissue non-suppurative inflammations are its main features. It becomes the second cause of loss ability to work behind the ischemic heart disease. The traditional treatment of RA drugs are mostly single pathway or target, while there are certain side effects, so development of efficient, low toxicity, the role of multi-target drug is particularly important and urgent.
Panax japonicus (Panax japonicum CA Mey.),a traditional chinese medical herb, belongs to Araliaceae. It has strong tonic, dissipate blood stasis and alleviate pain, bleeding and expectorant functions in Chinese medicine record.So it used totreated weakness after diseases, tuberculosis hemoptysis, cough, sputum, and bruises. In this study, its anti-inflammatory effects and anti-rheumatic mechanisms will be carried out syetemic researched.
1) The effect of TSPJ on normal inflammatory response in mice:
(1) TSPJ 50 mg/kg and 100 mg/kg group could significantly reduce the paw edema, foot redness swelling and other symptoms induced by carrageenan after inflammation 0.5～4 h (P<0.05)Vs control group.
(2) TSPJ could inhibit the mouse ear edema induced by croton oil, the ear's redness, swelling and other symptoms are significantly inhibited.The high dose group of TSPJ's inhibition rate is 33.33% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).
(3) TSPJ could inhibit the increased capillary permeability induced by acetic acid on mice inhibition rate of TSPJ's high dose group(100 mg/kg) is 48.6% compared with the control group. The inhibition rate is positively correlated with the dose (P<0.05).
(4) TSPJ could increasethe levels of serum GSH-PX, SOD activity and reduce serum MDA content. The inhibition rate is positively correlated with the dose
(5) TSPJ could reduce the inflammatory factors (IL-1, TNF-α) in mice serum, the differences are significant (P<0.05).
2) The effect of TSPJ on CIA rats'histopathological and ultrastructural stractures
(1) The AI scores in CIA group are higher than normal group, three dose group's AI of TSPJ were also increased compare with the normal group at the same period, but the difference were significant compare with the CIA group at the same period.
(2) The model group' feet swelling begins from d12 to the end, peak time was at d24-d28. The swelling degree of TSPJ groups were also increased compare with the normal group at the same period, but the difference was significant compare with the CIA group at the same period.
(3) CIA model rats'weight-gain rate slow down or stoped at the secondary inflammatory phase (after d18), TPT treatment group also shown the weight-gain rate, TSPJ groups' growth rate were normal.
(4) The immune function of CIA model could lead to higher compared with model group; the administration of TPT could decreasethe higher s spleen index and thymus index in CIA rats, the difference was significant,and TSPJ group alos decline differences but no significant.
(5) TSPJ can improve joint damage of CIA:thre dose groups of TSPJ can reduce the levels of congestion and edema of soft tissue joint, toe joint volume and scores; reduce synovial hyperemia, edema of the degree; significantly inhibit the CIA rats' synovial hyperplasia; inhibit the inflammatory cell' infiltration; inhibit chondrocytes proliferation.
(6) ConA-induced spleen T lymphocyte proliferation in CIA group was significantly lower than the control group, and IL-2 levels were also significantly lower, TSPJ groups can restore the CIA rat spleen lymphocytes and production of IL-2, but TPT group has no effect on proliferation of lymphocytes and production of IL-2.
(7) The productions of IL-1β, IL-6 and TNF-αwere significantly higher in CIA group's PMΦthan those of normal group (P<0.05), TSPJ medium and large doses groups can inhibit the production of proinflammatory cytokines IL-1β, IL-6 and TNF-αlevels in CIA's PMΦ.
(8) The synovial membrane ultrastructure:at the CIA group, the synovial cell lining layers were thickened up to 5-6 layers and disarrangement, organelle deformation, or even disappear, the cytoplasm filled with large vacuoles, the uneven distribution of nuclear chromatin. Some cells due to degeneration and necrosis to atrophy, only to see a large number of cell fragments with staggered collagen fibers. TSPJ can significantly improve the CIA synovial ultrastructure of synovial lining cell layer and cell arrangement, only occasionally a small amount of lymphocytic infiltration can be seen, A-type cells to primary lysosomes, mitochondria slightly increased, B-type Cytoplasm of cells rich in rough endoplasmic reticulum and ribosome bodies, various types of cells close to normal.
(9) The ICAM-1 production (the total number of positive cells, the average black level value of the total area and integral optical density) were significantly increased in CIA model group's synovial membrane during the inflammatory process, TSPJ groups can reducethe ICAM-1 expression in synovial number, size Sum, integral optical density and average black levels, the differences are significant (P<0.05).
(10) synovial cell culture and cytokine researches showed:normal synovial cells' cytokines productions are very low even no detected, cell growth rate are normal; after stimulated by LPS, the speed of cell proliferation and cytokines such as IL-1, IL-6, IL-8 and TNF-αsecretions are increased, this suggest that LPS is a good stimulant of microbial antigens, can induce normal cell proliferation and differentiation. CIA synovial cells' proliferation rate and cytokines secretions were significantly higher after LPS stimulation. Tripterygium showed a strong anti-inflammatory effects; TSPJ didn't show cell-damage in the synovial cell culture experiments,and didn't stimulate cytokine secretion, only regulate the CIA synovial cells from "super- sensitivity" status to normal.
From the experimental results, TSPJ has the following parameters:(1) increasing the antioxidant capacity, reduce inflammation, tissue damage induced by free radicals; (2) to relieve inflammatory mediators such as PGE and LTB in inflamed tissue; (3) significantly decreased cell factors such as IL-1B, IL-6, TNF-αinf lammated tissues; (4) reduce the ICAM-1 generation in inflammated tissues; (5) TSPJ has 'two-way adjustment' on the immune system, so at the process of treating RA it can reduce hyperthyroidism immune function to normal, but don't produce immune dysfunction as triptolide and other immunosuppressive agents do. Therefore, TSPJ is likely to be developed as an effective, non-toxic or low toxic anti-inflammatory and anti- rheumatic drug.
引文
[1]Choi Y., Arron J. R., Townsend M. Promising bone-related therapeutic targets for rheumatoid arthritis. J. Nat. Rev. Rheumatol,2009,5:543-548
[2]Khanna D., Sethi G, Ahn K. S., et al. Natural products as a gold mine for arthritis treatment. Cur. Opin. Pharmacol,2007,7:344-351
[3]Manning A. M., Davis R. Targeting JNK for therapeutic benefit:From junk to gold?. J. Nat. Rev. Drug Disc,2003,2:554-565
[4]RaiseLG. Prostaglandins and bone:physiology and pathophysiology. J. Osteoarthritis Cartilage,1999,7(4):419-21
[5]MarjolaineGosset, FraneisBerenbaum, ArletteLevy. Prostaglandin E2 synthesis in cartilage explants under compression mPGES-1 1s a mechanosensitive gene. Arthritis Res Ther,2006,8(11):R135
[6]SeottDL, PugnerK, Karrela K. The links between joint damage and disability in rheumatoid arthritis. J. Rheumatol,2000,39(2):122-32
[7]蒋明,林孝义,朱立平等主编.中华风湿病学.北京：华夏出版社,2004：737
[8]Lee D. M., Weinblatt M. E. Rheumatoid arthritis. Lancet,2001,358(9285):903-911
[9]白洪宇.大鼠类风湿性关节炎实验模型的建立与评价[D].2009
[10]Ono K., Han J. The p38 signal transduction pathway Activation and function Cellular Signalling,2000,12:1 (1-13)
[11]马丽辉.骨髓间充质干细胞治疗类风湿关节炎机理和相关研究[D].2008
[12]Okamoto H., Cujec T. P., Yamanaka H., et al. Molecular aspects of rheumatoid arthritis:Role of transcription factors. FEBS Journal 2008,275(18):4463-4470
[13]Cano E., Mahadevan L. C. Parallel signal processing among mammalian MAPKs. TIBS,1995,20:117
[14]Sweeney S. E., Kimbler T. B., Firestein G. S. Role of interferon regulatory factors in the synoviocyte type Ⅰ IFN response. Arthritis Rheum,2009,60(10):140-142
[15]Li Q., Verma I. M. Erratum:NF-κB regulation in the immune system. Nature Reviews Immunology,2002,2(12):975-979
[16]Dejardin E., Droin N. M., Delhase M., et al. The lymphotoxin-β receptor induces different patterns of gene expression via two NF-κB pathways. Immunity,2002, 17(4):525-535
[17]Ghosh S., Karin M. Missing pieces in the NF-κB puzzle. Cell,2002,109(2): S81-S96
[18]Silverman N., Maniatis T. NF-κB signaling pathways in mammalian and insect innate immunity. Genes and Development,2001,15(18):2321-2342
[19]Benbow U., Brinckerhoff C. E. The AP-1 site and MMP gene regulation:What is all the fuss about. Matrix Biol,1997,15:519-526
[20]Foster L. C., Wiesel P., Huggins G. S., et al. Role of activating protein-1 and high mobility group-I(Y) protein in the induction of CD44 gene expression by interleukin-1β in vascular smooth muscle cells. FASEB J.,2000,14:368-378
[21]Harrison L. M., Van Haaften W. C. E., Tesh V. L. Regulation of Proinflammatory Cytokine Expression by Shiga Toxin 1 and/or Lipopolysaccharides in the Human Monocytic Cell Line THP-1. Infect. Immun,2004,72:2618-2627
[22]Rooney J. W., Sun Y. A.-L., Glimcher L. H., et al. Novel NFAT sites that mediate activation of the interleukin-2 promoter in response to T-cell receptor stimulation. Mol. Cell. Biol.,1995,15:6299-6310
[23]Brogan I. J., Murray I. A., Cerillo G., et al. Interaction of glucocorticoid receptor isoforms with transcription factors AP-1 and NF-κB:Lack of effect of glucocorticoid receptor β. Mol. Cell. Endocrinol,1999,157:95-104
[24]Rincon M., Derijard B., Chow C. W., et al. Reprogramming the signalling requirement for AP-1 (activator protein-1) activation during differentiation of precursor CD4+ T-cells into effector Thl and Th2 cells. Genes Funct,1997,1: 51-68
[25]Sumiko W., Ken-ichi A. Roles of the JAK-STAT system in signal transduction via cytokine receptors. Cur Opin Gene Devel,1996,6(5):587-596
[26]Ihle J. N., Nosaka T., Thierfelder W., et al. Shimoda K. Jaks and Stats in cytokine signaling. Stem Cells,1997,15(suppl 1):105-12
[27]Kotlyarov A., Yannoni Y, Fritz S., et al. Distinct cellular functions of MK2. Mol. Cell. Biol.,2002,22:4827-4835
[28]Kontoyiannis D., Boulougouris G., Manoloukos M., et al. Genetic dissection of the cellular pathways and signaling mechanisms in modeled tumor necrosis factor-induced Crohn's-like inflammatory bowel disease. J. Exp. Med.,2002,196: 1563-1574
[29]Andersen T. L., Del Carmen Ovejero M., Kirkegaard T., et al. A scrutiny of matrix metalloproteinases in osteoclasts:Evidence for heterogeneity and for the presence of MMPs synthesized by other cells Bone,2004,35(5):1107-1119
[30]Tadashi Takahashi, Takao Nakamura, Atsushi Hayashi, et al. Inhibition of experimental choroidal neovascularization by overexpression of tissue inhibitor of metalloproteinase-3 in retinal pigment epithelium cells Original Research Article. Am J Ophthal,2000,130(6):774-781
[31]Raza S. L., Nehring L. C., Shapiro S. D., et al. Proteinase-activated receptor-1 regulation of macrophage elastase (MMP-12) secretion by serine proteinases. J. Bio. Chem.,2000,275:41243-41250
[32]Carmichael D. F., Stricklin G P., Stuart J. M. Systemic administration of TIMP in the treatment of collagen-induced arthritis in mice. Agents and Actions,1989,27: 378-379
[33]Mohammed F. F., Smookler D. S., Khokha R. Metalloproteinases, inflammation, and rheumatoid arthritis. Ann. Rheum. Dis.,2003,62:43-47
[34]Funk C. D. Prostaglandins and leukotrienes:Advances in eicosanoid biology. Science,2001,294:1871-1875
[35]Rivera R., Chun J. Biological effects of lysophospholipids. Rev. Physiol. Biochem. Pharmacol,2008,160:25-46
[36]Xu Y., Xiao Y. J., Zhu K., et al. Unfolding the pathophysiological role of bioactive lysophospholipids. Curr. Drug Targets Immune Endocr. Metabol. Disord,2003,3: 23-32
[37]Burke J. E., Dennis E. A. Phospholipase A2 structure/function, mechanism, and signaling. J. Lipid Res.,2009,50:S237-242
[38]Schaloske R. H., Dennis E. A. The phospholipase A2 superfamily and its group numbering system. Biochim. Biophys. Acta.,2006,1761:1246-1259
[39]Raichel L., Berger S., Hadad N., et al. Reduction of cPLA2a overexpression:An efficient anti-inflammatory therapy for collagen-induced arthritis. Eur. J. Immunol. 2008,38:2905-2915
[40]Clark J. D., Lin L.-L., Kriz R. W., et al. A novel arachidonic acid-selective cytosolic PLA2 contains a Ca2+-dependent translocation domain with homology to PKC and GAP. Cell,1991,65(6):1043-1051
[41]Ghosh M., Tucker D. E., Burchett S. A., et al. Properties of the Group IV phospholipase A2 family. Prog. Lipid Res.,2006,45:487-510
[42]Niknami M., Patel M., Witting P. K., et al. Molecules in focus:Cytosolic phospholipase A2-a. Int. J. Biochem. Cell Biol.,2009,41:994-997
[43]Honda T., Segi-Nishida E., Miyachi Y., et al. Prostacyclin-IP signaling and prostaglandin E2-EP2/EP4 signaling both mediate joint inflammation in mouse collagen-induced arthritis. J. Exp. Med.,2006:325-335
[44]Trebino C. E., Stock J. L., Gibbons C. P., et al. Impaired inflammatory and pain responses in mice lacking an inducible prostaglandin E synthase. Proc. Natl. Acad. Sci. U. S. A.,2003,100:9044-9049
[45]Lemos H. P., Grespan R., Vieira S. M., et al. Prostaglandin mediates IL-23/IL-17-induced neutrophil migration in inflammation by inhibiting IL-12 and IFNγ production. Proc. Natl. Acad. Sci. U. S. A.,2009,106:5954-5959
[46]Miwa M., Saura R., Hirata S., et al. Induction of apoptosis in bovine articular chondrocyte by prostaglandin E2 through c AMP-dependent pathway. Osteoarthritis Cartilage,2000,8:17-24
[47]Dogne J. M., De Leval X., Neven P., et al. Effects of a novel non-carboxylic thromboxane A2 receptor antagonist (BM-531) derived from torasemide on platelet function. Prostaglandins Leukot. Essent. Fatty Acids,2000,62:311-317
[48]Mathis S., Jala V. R., Haribabu B. Role of leukotriene B4 receptors in rheumatoid arthritis. Autoimmun. Rev.,2007,7:12-17
[49]Kronke G, Uderhardt S., Katzenbeisser J., et al. The 12/15-lipoxygenase pathway promotes osteoclast development and differentiation. Autoimmunity,2009,42: 383-385
[50]Chen Z. K., Lv H. S. Quantification of expression of leukotriene B4 inducing tumor necrosis factor-alpha and interleukin-lbeta at mRNA level in synovial membrane cells of rheumatoid arthritis by real-time quantitative PCR. J Peking Univ. Health sciences,2006,38:533-536
[51]Xu S., Lu H., Lin J., et al. Regulation of TNFa and IL1β in rheumatoid arthritis synovial fibroblasts by leukotriene B4. Rheumatol Int.,2010,30:1183-1189
[52]Wipke B. T., Allen P. M. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J. Immunol.,2001,167: 1601-1608
[53]Anderson G. D., Hauser S. D., McGarity K. L., et al. Selective inhibition of cyclooxygenase (COX)-2 reverses inflammation and expression of COX-2 and interleukin 6 in rat adjuvant arthritis. J. Clin. Invest.,1996,97:2672-2679
[54]Cortes-Burgos L. A., Zweifel B. S., Settle S. L., et al. CJ-13610, an orally active inhibitor of 5-lipoxygenase is efficacious in preclinical models of pain. Eur. J. Pharmacol.,2009,617:59-67
[55]Araico A., Terencio M. C., Alcaraz M. J., et al. Evaluation of the anti-inflammatory and analgesic activity of Me-UCH9, a dual cyclooxygenase-2/5-lipoxygenase inhibitor. Life Sci.,2007,80:2108-2117
[56]Martel-Pelletier J., Lajeunesse D., Reboul P., et al. Therapeutic role of dual inhibitors of 5-LOX and COX, selective and non-selective non-steroidal anti-inflammatory drugsAnn. Rheum. Dis.,2003,62:501-509
[57]Kehrer J. P. Free radicals as mediators of tissue injury and disease. Crit. Rev. Toxicol.,1993,23:21-48
[58]Merry P., Winyard P. G, Morris C. J., et al. Oxygen free radicals, inflammation, and synovitis:and synovitis:the current status. Ann. Rheum. Dis.,1989,48:864-870
[59]Cuzzocrea S. Role of nitric oxide and reactive oxygen species in arthritis. Curr. Pharm. Des.,2006,12:3551-3570
[60]Rostoka E., Baumane L., Isajevs S., et al. Effects of kaempferol and myricetin on inducible nitric oxide synthase expression and nitric oxide production in rats. Basic Clin. Pharmacol. Toxicol.,2010,106:461-466
[61]Evans C. H., Robbins P. D. Gene therapy of arthritis. Int Med,1999,38:233-239
[62]Muzzonigro T. S., Ghivizzani S. C., Robbins P. D., et al. The role of gene therapy. Fact or fiction. Clin Sports Med,1999,18:223-239
[63]Evans C. H., Gouze J. N., Gouze E., et al. Osteoarthritis gene therapy. Gene Ther, 2004,11:379-389
[64]Evans C. H., Robbins P. D. Potential treatment of osteoarthritis by gene therapy. Rheum Dis Clin North Am,1999,25:333-344
[65]高凯.腺相关病毒/sTNFRⅡ:Fc基因治疗类风湿关节炎的研究[D].2005
[66]陈昶.蓬草除痹汤对CIA大鼠滑膜炎及骨浸蚀作用机制的实验研究[D].2010
[67]吴素玲.痹痛灵颗粒对类风湿性关节炎关节滑膜保护机理的研究[D].2010
[68]李晋奇.制川乌与白芍组分配伍治疗类风湿性关节炎机理的研究[D].2008
[69]邓长财.痹清饮治疗类风湿性关节炎的临床及对滑膜细胞的干预作用[D].2008
[70]国家药典委员会.中国药典.Ⅰ部[S].北京：化学工业出版社,2000：106-107
[71]张颖.关于竹节三七中皂苷组成的分析[J].国外医学·中医中药分册,1997,19(1)：45-46
[72]左锐,袁丁.竹节参化学成分和药理活性研究进展[J].时珍国医国药,2005,16(9)：838-840
[73]闵静,黄琼,景玉萍等.竹节三七皂苷治疗风湿性疾病实验研究[J].湖北职业技术学院学报,2010,13(2)：104-106
[74]闵静,景玉萍,刘慧明等.竹节参总皂苷对类风湿性关节炎大鼠的抗炎作用及其机制研究[J].咸宁学院学报(医学版),2011,25(4)：280-282
[75]陈淑清,余立江,江帆.竹节三七总皂苷的镇痛,镇静,抗惊及解痉作用和毒性试验[J].华西药学杂志,1987,2(2)：85
[76]鄢丰,杨频.从竹节三七,肉桂,大黄中提取的天然活性成分的镇痛作用研究[J].中国新医药,2003,2(5)：21-22
[77]Zou K., Zhu S., Meselhy M. R. Dammarane-type saponins for Panax japonicus and their neurite out growth activity in SKN-SH cells. J nat. prod.,2002,65(9): 1288-1289
[78]李君庆.年龄及人参皂苷Rg1对大鼠大脑皮层NO释放的影响[J].药学学报,1997,32(6)：63-64
[79]张宝恒,齐治,温淑荣.竹节人参提取物对心血管的作用及对cAMP和cGMP 含量的影响[J].中心血管病杂志,1986,14(2)：100
[80]刘家兰.竹节人参提取物对实验性心肌缺血的保护作用[J].湖北民族学院学报,1999,16(1)：626
[81]Matsuda H., Samukawa K., Fukuda S1. Studies of Panax japonicus fibrinolysis. Planta medica,1989,55(1):18
[82]陈龙全,郜红利.复方竹节三七片对大鼠佐剂性关节炎治疗作用的实验研究[J].中国实验方剂学杂志,2003：(3)：25-25
[83]闵静,敖明章,刘慧明等.竹节参总皂苷对免疫器官的影响[J].湖北职业技术学院学报,2008,11(1)：109-111
[84]Hasegawa H., Matsumiya S., Uchiyama M. Inhibitory effect of some triterpenoid saponins on glucose transport in tumor cells and its application to in vitro cytotoxic and antiviral activities. Planta medica,1994,60(3):240-243
[85]张国斌.蜈蚣三七提取物抗类风湿性关节炎免疫机理的研究[D].2006
[86]孟卫星,陈龙全,袁德培.复方竹节三七对小鼠免疫功能影响的实验研究[J].恩施医专学报,1998,15(4)：123
[87]成龙全,陈朝俊.复方竹节三七片抗风湿作用的免疫调节机制研究[J].中国医药学报,2003,18(4)：247-248
[88]陈奇修.中药药理实验方法学[M].第1版.北京：人民卫生出版社,1993：317
[89]徐叔云,卞如濂,陈修.药理实验方法学[M].第3版.北京：人民卫生出版社,2002：1420-1455
[90]莫选荣罗心静类风湿性关节炎发病机制的研究进展[J].医学临床研究,2007,24：337-340
[91]李玉洁,王强状,郑晓俊.雷公藤治疗类风湿性关节炎的机制研究现状.中国药师,2004,7(8)：639-641
[92]吴柏林.雷公藤治疗类风湿性关节炎的机理探要.实用中医内科杂志,2003,17(3)：150
[93]Cibere J., Deng Z., Lin Y. Randomized double-blind Placebo-controlled study of Tripterygium wilfordii, a Chinese herbal therapy in rheumatoid arthritis. Aithritis Rheum.2000:43(1):1345
[94]Asano K., Yu Y., Kasahara T. Inhibition of murine chronic graft-versus-hos t disease by the chloroform extract of Tripterygium wilfordii Hook f. Transpl Immunol,1997, 5:315
[95]Qiu D., Kao P. N. Immunosuppressive and anti-inflammatory mechanisms of triptolide, the principal diterpenoid from the Chinese medicinal herb Tripterygium wilfordii Hook. f. DrugsR&D.,2003,4(1):1-18
[96]喻丽珍,卞慧敏.雷公藤毒副作用的研究进展.黑龙江中医药,2005(02)：52-53
[97]张季.严春临中药雷公藤甲素药理学和毒理学研究进展.内蒙古中医药,2006(6)：55-57
[98]刘琼,王晨瑶.类风湿性关节炎动物模型的研究进展[J].医学综述,2006,12(5)：313-316
[99]Andrew D. Cook, Merrill J. Rowley, Bruce D. Wines, et al. Antibodies to the Collagen-like Region of C1q and Type Ⅱ Collagen are Independent Non-cross-reactive Populations in Systemic Lupus Erythematosus and Rheumatoid Arthritis. J Autoimmu,1994,7:369-378
[100]George R. Dodge, Isabelle Pidoux, A. Robin Poole. The Degradation of Type Ⅱ Collagen in Rheumatoid Arthritis:An Immunoelectron Microscopic Study. Matrix, 1991,11:330-338
[101]Cutolo M., Accuado S. Sex hormones, HLA and the rheumatoid arthritis. J Clin Exp Rheumat,1991,9:641-642
[102]艾景录,吕爱平.性腺功能异常型肾虚痹证发生机理的实验研究[J].中国中医基础医学杂志,1995,2：142-145
[103]Ostensen M., Veiby O. P., Raiss R. Reponses of normal and rheumatoid human anticular chondrocyte cultured under various experimental conditions in agarose. Scond J Rheumatol,1991,20:172-174
[104]吕爱平,王安民,曾晓莲.益肾镯痹丸对大鼠实验性痹证影响的病理学研究[J].中医杂志,1989,29：49-51
[105]Clague R. B., Moore L. J. IgG and IgM antibody to native type Ⅱ collagen in the arthritis serum and synovial fluid. Arthritis Rheum,1984,27:1370-1376
[106]Shinmei M., Masuda K., Kikuchi T. Produtions of cytokines by chondrocytes and its role in proteoglycan degradation. J Rheumatol supl,1991,27:89-91
[107]Shuckett R., Malemud C. J. Proteoglycans synthesized by chondrocytes of human nonarthritie and osteoarthritic cartilage. Proc Soc Exp biomed,1989,110:275-279
[108]Akaogi J., Nozaki T., Satoh M., et al. Role of PGE2 and EP receptors in the pathogenesis of rheumatoid arthritis and as a novel therapeutic strategy. Endocr. Metab. Immune Disord. Drug Targets,2006,6(4):383-394
[109]Ghosh M., Tucker D. E., Burchett S. A., et al. Properties of the Group Ⅳ phospholipase A2 family. Prog. Lipid Res.,2006,45(6):487-510
[110]Niknami M., Patel M., Witting P. K., et al. Molecules in focus:Cytosolic phospholipase A2-α. Int. J. Biochem. Cell Biol.,2009,41:994-997
[111]Uozumi N., Shimizu T. Roles for cytosolic phospholipase A2α as revealed by gene-targeted mice. Prostaglandins and Other Lipid Mediators,2002,68-69:59-69
[112]Brenner T., Brocke S., Szafer F., et al. Inhibition of nitric oxide synthase for treatment of experimental autoimmune encephalomyelitis. J. Immunol.,1997, 158(6):2940-2946
[113]Miwa M., Saura R., Hirata S., et al. Induction of apoptosis in bovine articular chondrocyte by prostaglandin E2 through cAMP-dependent pathway. Osteoarthritis Cartilage,2000,8(1):17-24
[114]Pengfei Li, James A. Cook, Gary S. Gilkeson, et al, Hongkuan Fan Increased expression of beta-arrestin 1 and 2 in murine model of RA:Isoform specific regulation of inflammation. Mol Immun.,2011,49:64-74
[115]Daniel T. O., Liu H., Morrow J. D., et al. Thromboxane A2 is a mediator of cyclooxygenase-2-dependent endothelial migration and angiogenesis. Cancer Res, 1999,59(18):4574-4577
[116]Mathis S., Jala V. R., Haribabu B. Role of leukotriene B4 receptors in rheumatoid arthritis. Autoimmun. Rev.,2007,7(1):12-17
[117]Lemos H. P., Grespan R., Vieira S. M., et al. Prostaglandin mediates IL-23/IL-17-induced neutrophil migration in inflammation by inhibiting IL-12 and IFNγ production. Proc. Natl. Acad. Sci. U. S. A.,2009,106:5954-5959
[118]Andersen T. L., Del Carmen Ovejero M., Kirkegaard T., et al. A scrutiny of matrix metalloproteinases in osteoclasts:Evidence for heterogeneity and for the presence of MMPs synthesized by other cells. Bone,2004,35 (5):1107-1119
[119]Yaqin Yu, Ran Tao, Jieping Shi, et al. A genetic study of two calcium-independent cytosolic PLA2 genes in schizophrenia. Prosta Leukot Essent Fatty Acids,2005,73: 351-354
[120]Raza S. L., Cornelius L. A. Matrix metalloproteinases:Pro- and anti-angiogenic activities J. Invest. Dermatol. Symp. Proc.,2000,5:47
[121]Fallarino F., Grohmann U., Hwang K. W., et al. Modulation of tryptophan catabolism by regulatory T cells. Nat. Immunol.,2003,4(12):1206-1212
[122]Emmanouil Galanakis, Diamantis Kofteridis, Kalliopi Stratigi, et al. Intron 4 a/b polymorphism of the endothelial NOS gene is associated with both type 1 and type2 diabetes in a genetically homogeneous population. Human Immunology,2008,69: 279-283
[123]DeSilva D. R., Jones E. A., Favata M. F., et al. Inhibition of mitogen-activated protein kinase kinase blocks T cell proliferation but does not induce or prevent anergy. J Immunol,1998,160(9):4175-4181
[124]Moses M. A. The regulation of neovascularization by matrix metalloproteinases and their inhibitors. Stem Cells,1997,15(3):180-189
[125]Hayashi T., Beck L., Rossetto C., et al. Inhibition of experimental asthma by indoleamine 2,3-dioxygenase. J Clin. Invest.,2004,114(2):270-279
[126]Criado G., Simelyte E., Inglis J. J., et al. Indoleamine 2,3 dioxygenase-mediated tryptophan catabolism regulates accumulation of Thl/Thl7 cells in the joint in collagen-induced arthritis. Arthritis Rheuma,2009,60(5):1342-1351
[127]Tuyt L. M., Dokter W. H., Birkenkamp K., et al. Extracellular-regulated kinase 1/2, Jun N-terminal kinase, and c-Jun are involved in NF-κB-dependent IL-6 expression in human monocytes. J. Immunol.,1999,162:4893-4902
[128]Pages G., Guerin S., Grall D., et al. Defective thymocyte maturation in p44 MAP kinase (Erk 1) knockout mice. Science,1999,286(5443):1374-1377
[129]Szanto S., Koreny T., Mikecz K., et al. Inhibition of indoleamine 2,3-dioxygenase-mediated tryptophan catabolism accelerates collagen-induced arthritis in mice. Arthritis res. Ther.,2007,9(3):R50
[130]Schett G, Tohidast-Akrad M., Smolen J. S., et al. Activation, differential localization and regulation of the stress-activated protein kinases, extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase, in synovial tissue and cells in rheumatoid arthritis. Arthritis Rheuma,2000, 43(11):2501-2512
[131]Scherle P. A., Jones E. A., Favata M. F., et al. Inhibition of MAP kinase kinase prevents cytokine and prostaglandin E2 production in lipopolysaccharide-stimulated monocytes. J Immunol.,1998,161(10):5681-5686
[132]Brogley M. A., Cruz M., Cheung H. S. Basic calcium phosphate crystal induction of collagenase 1 and stromelysin expression is dependent on a p42/44 mitogen-activated protein kinase signal transduction pathway. J. Cell Physiol.1999,180: 215-224
[133]Benbow U., Brinckerhoff C. E. The AP-1 site and MMP gene regulation:What is all the fuss about. Matrix Biol,1997,15:519-526
[134]Shi Y., Gaestel M. In the cellular garden of forking paths:How p38 MAPKs signal for downstream assistance. Bio. Chem.,2002,383:1519-1536
[135]Udalova I. A., Mott R., Field D., et al. Quantitative prediction of NF-κB DNA-protein interactions. Proc. Nat. Aca. Sci. U. S. A.,2002,99:8167-8172
[136]Harrison L. M., Van Haaften W. C. E., Tesh V. L. Regulation of Proinflammatory Cytokine Expression by Shiga Toxin 1 and/or Lipopolysaccharides in the Human Monocytic Cell Line THP-1. Infect. Immun,2004,72:2618-2627
[137]Foster L. C., Wiesel P., Huggins G. S., et al. Role of activating protein-1 and high mobility group-Ⅰ(Y) protein in the induction of CD44 gene expression by interleukin-1β in vascular smooth muscle cells. FASEB J.,2000,14:368-378
[138]Ono K., Han J. The p38 signal transduction pathway Activation and function. Cell. Signal,2000,12:1-13
[139]Mamani-Matsuda M., Kauss T., Al-Kharrat A., et al. Therapeutic and preventive properties of quercetin in experimental arthritis correlate with decreased macrophage inflammatory mediators. Biochemi. Pharma.,2006,72:1304-1310
[140]Nataraju A., Raghavendra Gowda C. D., Rajesh R., et al. Group IIA secretory PLA2 inhibition by ursolic acid:A potent anti-inflammatory molecule. Cur. Top. Med. Chem.,2007,7:801-809
[141]Takada Y., Ichikawa H., Badmaev V., et al. Acetyl-11-keto-β-boswellic acid potentiates apoptosis, inhibits invasion, and abolishes osteoclastogenesis by suppressing NF-κB and NF-κB-regulated gene expression. J Immunol,2006,176: 3127-3140
[142]Kimmatkar N., Thawani V., Hingorani L., et al. Efficacy and tolerability of Boswellia serrata extract in treatment of osteoarthritis of knee-A randomized double blind placebo controlled trial. Phytomed,2003,10:3-7
[143]Adcocks C,. Collin P., Buttle D. J. Catechins from green tea (Camellia sinensis) inhibit bovine and human cartilage proteoglycan and type Ⅱ collagen degradation in vitro. J Nutri.,2002,132:341-346
[144]Funk J. L., Frye J. B., Oyarzo J. N., et al. Efficacy and mechanism of action of turmeric supplements in the treatment of experimental arthritis. Arthritis Rheum., 2006,54:3452-3464
[145]曾克勤,胡永红,张明敏等.雷公藤甲素对CIA大鼠T细胞胞内细胞因子表达的影响[J].北京中医药大学学报,2004,27(5)：39
[146]Yu F., Yu F., Li R., Wang R. Inhibitory effects of the Gentiana macrophylla (Gentianaceae) extract on rheumatoid arthritis of rats. J Ethnopharmaco.,2004,95: 77-81
[147]Kim S. M, Lee S. Y, Yuk D. Y, et al. Inhibition of NF-kappaB by ginsenoside Rg3 enhances the susceptibility of colon cancer cells to docetaxel. Arch. Pharmacal. Res., 2009,32:755-765
[148]Yang D. S., Liu F., Zeng F. D., et al. Effect of sinomenine on adjuvant arthritis and its mechanisms. China j Chin materia med,2005,30:1361-1363
[149]Wang Y, Jia L., Wu C.-Y. Triptolide inhibits the differentiation of Th17 cells and suppresses collagen-induced arthritis. Scand. J Immunol,2008,68:383-390
[150]Sun B., Lv L., Lu Z.-X., et al. Study on drug-guide effect of Achyranthes bidentata in Sanmiao pill in arthritic rats. Zhongguo Zhongyao Zazhi,2008,33:2946-2949
[151]Roy D. Altman The future of herbal medicine in the treatment of osteoarthritis. Osteoarthritis Cartilage,2008,16:S10-S11
[152]Zhang L., Li J., Yu S.-C., et al. Therapeutic effects and mechanisms of total flavonoids of Turpinia Arguta Seen on adjuvant arthritis in rats. J Ethnopharmacol., 2008,116:167-172
[153]Dinesh Khanna, Gautam Sethi, Kwang Seok Ahn, et al. Amita Aggarwal and Bharat B Aggarwal. Natural products as a gold mine for arthritis treatment. Curr Opin in Pharmacol,2007,7:344-351
[154]Aggarwal B. B., Bhardwaj A., Aggarwal R. S., et al. Role of resveratrol in prevention and therapy of cancer:Preclinical and clinical studies. Anticancer Res., 2004,24:2783-2840
[155]Funk J. L., Frye J. B., Oyarzo J. N., et al. Comparative effects of two gingerol-containing zingiber officinale extracts on experimental Rheumatoid arthritis. J Nat. Prod.,2009,72:403-407
[156]Yoshida N., Kanekura T., Higashi Y., et al. Bidens pilosa suppresses interleukin-1β-induced cyclooxygenase-2 expression through the inhibition of mitogen activated protein kinases phosphorylation in normal human dermal fibroblasts. J Dermatol.,2006,33:676-683
[157]Xiaoyan Shen, Cuixian Li, Huifang Zhao, et al. Inhibitory effects of a traditional Chinese herbal formula TBL-Ⅱ on type Ⅱ collagen-induced arthritis in mice. J Ethnopharmacol,2011,134(2):399-405
[158]Hwang E.-S., Park K.-K. Magnolol suppresses metastasis via inhibition of invasion, migration, and matrix metalloproteinase-2/-9 activities in PC-3 human prostate carcinoma cells. Biosci, Biotech. Biochem.,2010,74:961-967
[159]Ahmad S. F., Khan B., Bani S., et al. Amelioration of adjuvant-induced arthritis by ursolic acid through altered Th1/Th2 cytokine production. Pharmacol. Res.,2006, 53:233-240
[160]Kiani A., Rao A., Aramburu J. Manipulating immune responses with immunosuppressive agents that target NFAT. Immunity,2000,12:359-372
[161]Ramgolam V., Ang S. G., Lai Y. H., et al. Traditional Chinese medicines as immunosuppressive agents. Ann. Aca. Med. e Singapore.,2000,29:11-16
[162]Ou Y, Li W., Li X., et al. Sinomenine reduces invasion and migration ability in fibroblast-like synoviocytes cells co-cultured with activated human monocytic THP-1 cells by inhibiting the expression of MMP-2, MMP-9, CD147. Rheuma. Int., 2010(1-7)
[163]Rostoka E., Baumane L., Isajevs S., et al. Effects of kaempferol and myricetin on inducible nitric oxide synthase expression and nitric oxide production in rats. Basic Clinic. Pharmacol. Toxicol.,2010,106:461-466
[164]Tong L., Moudgil K. D. Celastrus aculeatus Merr. suppresses the induction and progression of autoimmune arthritis by modulating immune response to heat-shock protein 65. Arthritis res. Ther.,2007,9:4 (R70)
[165]Sindhu G, Ratheesh M., Shyni G. L., et al. Inhibitory effects of Cynodon dactylon L. on inflammation and oxidative stress in adjuvant treated rats. Immunopharmacol. Immunotoxicol,2009,31:647-653
[166]Yang D. S., Liu F., Zeng F. D., et al. Effect of sinomenine on adjuvant arthritis and its mechanisms. Zhongguo Zhong yao Za zhi,2005,30:1361-1363
[167]Koprowska K., Czyz M. Molecular mechanisms of parthenolide's action:Old drug with a new face. Postepy Hig. Med. Dosw (Online) 2010,64:100-114
[168]Yu S., Zheng W., Xin N., Chi Z.-H., Wang N.-Q., Nie Y.-X., Feng W.-Y., Wang Z.-Y. Curcumin prevents dopaminergic neuronal death through inhibition of the c-Jun N-terminal kinase pathway. Rejuvenation Res.2010,13:55-64
[169]Choi H. J., Park Y. R., Nepal M., et al. Inhibition of osteoclastogenic differentiation by Ikarisoside A in RAW 264.7 cells via JNK and NF-KB signaling pathways. Eur. J. Pharmacol.,2010,636:28-35
[170]Matta R., Wang X., Ge H., et al. Triptolide induces anti-inflammatory cellular responses. Am. J. Transl. Res.,2009,1:267-282
[171]Hsu YL., Chen CY, Hou MF., et al.6-dehydrogingerdione, an active constituent of dietary ginger, induces cell cycle arrest and apoptosis through reactive oxygen species/c-Jun N-terminal kinase pathways in human breast cancer cells. Mol. Nutr. Food Res.,2010,54:1307-1317
[172]Elmali N., Esenkaya I., Harma A., et al. Effect of resveratrol in experimental osteoarthritis in rabbits. Inflamma. Res.,2005,54:158-162
[173]Yang X., Liu Y, Bao Z., et al. Simultaneous determination of three active compounds in root barks of Dictamnus dasycarpus by RP-HPLC. Zhongguo Zhongyao Zazhi,2010,35:187-190
[174]Sethi G, Kwang S. A., Pandey M. K., et al. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-KB-regulated gene products and TAK1-mediated NF-κB activation. Blood., 2007,109:2727-2735
[175]Li H., Jia Y.-F., Pan Y, et al. Effect of tripterine on collagen-induced arthritis in rats. Acta Pharmacol. Sinica.,1997,18:270-273
[2]Khanna D., Sethi G, Ahn K. S., et al. Natural products as a gold mine for arthritis treatment. Cur. Opin. Pharmacol,2007,7:344-351
[3]Manning A. M., Davis R. Targeting JNK for therapeutic benefit:From junk to gold?. J. Nat. Rev. Drug Disc,2003,2:554-565
[4]RaiseLG. Prostaglandins and bone:physiology and pathophysiology. J. Osteoarthritis Cartilage,1999,7(4):419-21
[5]MarjolaineGosset, FraneisBerenbaum, ArletteLevy. Prostaglandin E2 synthesis in cartilage explants under compression mPGES-1 1s a mechanosensitive gene. Arthritis Res Ther,2006,8(11):R135
[6]SeottDL, PugnerK, Karrela K. The links between joint damage and disability in rheumatoid arthritis. J. Rheumatol,2000,39(2):122-32
[7]蒋明,林孝义,朱立平等主编.中华风湿病学.北京：华夏出版社,2004：737
[8]Lee D. M., Weinblatt M. E. Rheumatoid arthritis. Lancet,2001,358(9285):903-911
[9]白洪宇.大鼠类风湿性关节炎实验模型的建立与评价[D].2009
[10]Ono K., Han J. The p38 signal transduction pathway Activation and function Cellular Signalling,2000,12:1 (1-13)
[11]马丽辉.骨髓间充质干细胞治疗类风湿关节炎机理和相关研究[D].2008
[12]Okamoto H., Cujec T. P., Yamanaka H., et al. Molecular aspects of rheumatoid arthritis:Role of transcription factors. FEBS Journal 2008,275(18):4463-4470
[13]Cano E., Mahadevan L. C. Parallel signal processing among mammalian MAPKs. TIBS,1995,20:117
[14]Sweeney S. E., Kimbler T. B., Firestein G. S. Role of interferon regulatory factors in the synoviocyte type Ⅰ IFN response. Arthritis Rheum,2009,60(10):140-142
[15]Li Q., Verma I. M. Erratum:NF-κB regulation in the immune system. Nature Reviews Immunology,2002,2(12):975-979
[16]Dejardin E., Droin N. M., Delhase M., et al. The lymphotoxin-β receptor induces different patterns of gene expression via two NF-κB pathways. Immunity,2002, 17(4):525-535
[17]Ghosh S., Karin M. Missing pieces in the NF-κB puzzle. Cell,2002,109(2): S81-S96
[18]Silverman N., Maniatis T. NF-κB signaling pathways in mammalian and insect innate immunity. Genes and Development,2001,15(18):2321-2342
[19]Benbow U., Brinckerhoff C. E. The AP-1 site and MMP gene regulation:What is all the fuss about. Matrix Biol,1997,15:519-526
[20]Foster L. C., Wiesel P., Huggins G. S., et al. Role of activating protein-1 and high mobility group-I(Y) protein in the induction of CD44 gene expression by interleukin-1β in vascular smooth muscle cells. FASEB J.,2000,14:368-378
[21]Harrison L. M., Van Haaften W. C. E., Tesh V. L. Regulation of Proinflammatory Cytokine Expression by Shiga Toxin 1 and/or Lipopolysaccharides in the Human Monocytic Cell Line THP-1. Infect. Immun,2004,72:2618-2627
[22]Rooney J. W., Sun Y. A.-L., Glimcher L. H., et al. Novel NFAT sites that mediate activation of the interleukin-2 promoter in response to T-cell receptor stimulation. Mol. Cell. Biol.,1995,15:6299-6310
[23]Brogan I. J., Murray I. A., Cerillo G., et al. Interaction of glucocorticoid receptor isoforms with transcription factors AP-1 and NF-κB:Lack of effect of glucocorticoid receptor β. Mol. Cell. Endocrinol,1999,157:95-104
[24]Rincon M., Derijard B., Chow C. W., et al. Reprogramming the signalling requirement for AP-1 (activator protein-1) activation during differentiation of precursor CD4+ T-cells into effector Thl and Th2 cells. Genes Funct,1997,1: 51-68
[25]Sumiko W., Ken-ichi A. Roles of the JAK-STAT system in signal transduction via cytokine receptors. Cur Opin Gene Devel,1996,6(5):587-596
[26]Ihle J. N., Nosaka T., Thierfelder W., et al. Shimoda K. Jaks and Stats in cytokine signaling. Stem Cells,1997,15(suppl 1):105-12
[27]Kotlyarov A., Yannoni Y, Fritz S., et al. Distinct cellular functions of MK2. Mol. Cell. Biol.,2002,22:4827-4835
[28]Kontoyiannis D., Boulougouris G., Manoloukos M., et al. Genetic dissection of the cellular pathways and signaling mechanisms in modeled tumor necrosis factor-induced Crohn's-like inflammatory bowel disease. J. Exp. Med.,2002,196: 1563-1574
[29]Andersen T. L., Del Carmen Ovejero M., Kirkegaard T., et al. A scrutiny of matrix metalloproteinases in osteoclasts:Evidence for heterogeneity and for the presence of MMPs synthesized by other cells Bone,2004,35(5):1107-1119
[30]Tadashi Takahashi, Takao Nakamura, Atsushi Hayashi, et al. Inhibition of experimental choroidal neovascularization by overexpression of tissue inhibitor of metalloproteinase-3 in retinal pigment epithelium cells Original Research Article. Am J Ophthal,2000,130(6):774-781
[31]Raza S. L., Nehring L. C., Shapiro S. D., et al. Proteinase-activated receptor-1 regulation of macrophage elastase (MMP-12) secretion by serine proteinases. J. Bio. Chem.,2000,275:41243-41250
[32]Carmichael D. F., Stricklin G P., Stuart J. M. Systemic administration of TIMP in the treatment of collagen-induced arthritis in mice. Agents and Actions,1989,27: 378-379
[33]Mohammed F. F., Smookler D. S., Khokha R. Metalloproteinases, inflammation, and rheumatoid arthritis. Ann. Rheum. Dis.,2003,62:43-47
[34]Funk C. D. Prostaglandins and leukotrienes:Advances in eicosanoid biology. Science,2001,294:1871-1875
[35]Rivera R., Chun J. Biological effects of lysophospholipids. Rev. Physiol. Biochem. Pharmacol,2008,160:25-46
[36]Xu Y., Xiao Y. J., Zhu K., et al. Unfolding the pathophysiological role of bioactive lysophospholipids. Curr. Drug Targets Immune Endocr. Metabol. Disord,2003,3: 23-32
[37]Burke J. E., Dennis E. A. Phospholipase A2 structure/function, mechanism, and signaling. J. Lipid Res.,2009,50:S237-242
[38]Schaloske R. H., Dennis E. A. The phospholipase A2 superfamily and its group numbering system. Biochim. Biophys. Acta.,2006,1761:1246-1259
[39]Raichel L., Berger S., Hadad N., et al. Reduction of cPLA2a overexpression:An efficient anti-inflammatory therapy for collagen-induced arthritis. Eur. J. Immunol. 2008,38:2905-2915
[40]Clark J. D., Lin L.-L., Kriz R. W., et al. A novel arachidonic acid-selective cytosolic PLA2 contains a Ca2+-dependent translocation domain with homology to PKC and GAP. Cell,1991,65(6):1043-1051
[41]Ghosh M., Tucker D. E., Burchett S. A., et al. Properties of the Group IV phospholipase A2 family. Prog. Lipid Res.,2006,45:487-510
[42]Niknami M., Patel M., Witting P. K., et al. Molecules in focus:Cytosolic phospholipase A2-a. Int. J. Biochem. Cell Biol.,2009,41:994-997
[43]Honda T., Segi-Nishida E., Miyachi Y., et al. Prostacyclin-IP signaling and prostaglandin E2-EP2/EP4 signaling both mediate joint inflammation in mouse collagen-induced arthritis. J. Exp. Med.,2006:325-335
[44]Trebino C. E., Stock J. L., Gibbons C. P., et al. Impaired inflammatory and pain responses in mice lacking an inducible prostaglandin E synthase. Proc. Natl. Acad. Sci. U. S. A.,2003,100:9044-9049
[45]Lemos H. P., Grespan R., Vieira S. M., et al. Prostaglandin mediates IL-23/IL-17-induced neutrophil migration in inflammation by inhibiting IL-12 and IFNγ production. Proc. Natl. Acad. Sci. U. S. A.,2009,106:5954-5959
[46]Miwa M., Saura R., Hirata S., et al. Induction of apoptosis in bovine articular chondrocyte by prostaglandin E2 through c AMP-dependent pathway. Osteoarthritis Cartilage,2000,8:17-24
[47]Dogne J. M., De Leval X., Neven P., et al. Effects of a novel non-carboxylic thromboxane A2 receptor antagonist (BM-531) derived from torasemide on platelet function. Prostaglandins Leukot. Essent. Fatty Acids,2000,62:311-317
[48]Mathis S., Jala V. R., Haribabu B. Role of leukotriene B4 receptors in rheumatoid arthritis. Autoimmun. Rev.,2007,7:12-17
[49]Kronke G, Uderhardt S., Katzenbeisser J., et al. The 12/15-lipoxygenase pathway promotes osteoclast development and differentiation. Autoimmunity,2009,42: 383-385
[50]Chen Z. K., Lv H. S. Quantification of expression of leukotriene B4 inducing tumor necrosis factor-alpha and interleukin-lbeta at mRNA level in synovial membrane cells of rheumatoid arthritis by real-time quantitative PCR. J Peking Univ. Health sciences,2006,38:533-536
[51]Xu S., Lu H., Lin J., et al. Regulation of TNFa and IL1β in rheumatoid arthritis synovial fibroblasts by leukotriene B4. Rheumatol Int.,2010,30:1183-1189
[52]Wipke B. T., Allen P. M. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J. Immunol.,2001,167: 1601-1608
[53]Anderson G. D., Hauser S. D., McGarity K. L., et al. Selective inhibition of cyclooxygenase (COX)-2 reverses inflammation and expression of COX-2 and interleukin 6 in rat adjuvant arthritis. J. Clin. Invest.,1996,97:2672-2679
[54]Cortes-Burgos L. A., Zweifel B. S., Settle S. L., et al. CJ-13610, an orally active inhibitor of 5-lipoxygenase is efficacious in preclinical models of pain. Eur. J. Pharmacol.,2009,617:59-67
[55]Araico A., Terencio M. C., Alcaraz M. J., et al. Evaluation of the anti-inflammatory and analgesic activity of Me-UCH9, a dual cyclooxygenase-2/5-lipoxygenase inhibitor. Life Sci.,2007,80:2108-2117
[56]Martel-Pelletier J., Lajeunesse D., Reboul P., et al. Therapeutic role of dual inhibitors of 5-LOX and COX, selective and non-selective non-steroidal anti-inflammatory drugsAnn. Rheum. Dis.,2003,62:501-509
[57]Kehrer J. P. Free radicals as mediators of tissue injury and disease. Crit. Rev. Toxicol.,1993,23:21-48
[58]Merry P., Winyard P. G, Morris C. J., et al. Oxygen free radicals, inflammation, and synovitis:and synovitis:the current status. Ann. Rheum. Dis.,1989,48:864-870
[59]Cuzzocrea S. Role of nitric oxide and reactive oxygen species in arthritis. Curr. Pharm. Des.,2006,12:3551-3570
[60]Rostoka E., Baumane L., Isajevs S., et al. Effects of kaempferol and myricetin on inducible nitric oxide synthase expression and nitric oxide production in rats. Basic Clin. Pharmacol. Toxicol.,2010,106:461-466
[61]Evans C. H., Robbins P. D. Gene therapy of arthritis. Int Med,1999,38:233-239
[62]Muzzonigro T. S., Ghivizzani S. C., Robbins P. D., et al. The role of gene therapy. Fact or fiction. Clin Sports Med,1999,18:223-239
[63]Evans C. H., Gouze J. N., Gouze E., et al. Osteoarthritis gene therapy. Gene Ther, 2004,11:379-389
[64]Evans C. H., Robbins P. D. Potential treatment of osteoarthritis by gene therapy. Rheum Dis Clin North Am,1999,25:333-344
[65]高凯.腺相关病毒/sTNFRⅡ:Fc基因治疗类风湿关节炎的研究[D].2005
[66]陈昶.蓬草除痹汤对CIA大鼠滑膜炎及骨浸蚀作用机制的实验研究[D].2010
[67]吴素玲.痹痛灵颗粒对类风湿性关节炎关节滑膜保护机理的研究[D].2010
[68]李晋奇.制川乌与白芍组分配伍治疗类风湿性关节炎机理的研究[D].2008
[69]邓长财.痹清饮治疗类风湿性关节炎的临床及对滑膜细胞的干预作用[D].2008
[70]国家药典委员会.中国药典.Ⅰ部[S].北京：化学工业出版社,2000：106-107
[71]张颖.关于竹节三七中皂苷组成的分析[J].国外医学·中医中药分册,1997,19(1)：45-46
[72]左锐,袁丁.竹节参化学成分和药理活性研究进展[J].时珍国医国药,2005,16(9)：838-840
[73]闵静,黄琼,景玉萍等.竹节三七皂苷治疗风湿性疾病实验研究[J].湖北职业技术学院学报,2010,13(2)：104-106
[74]闵静,景玉萍,刘慧明等.竹节参总皂苷对类风湿性关节炎大鼠的抗炎作用及其机制研究[J].咸宁学院学报(医学版),2011,25(4)：280-282
[75]陈淑清,余立江,江帆.竹节三七总皂苷的镇痛,镇静,抗惊及解痉作用和毒性试验[J].华西药学杂志,1987,2(2)：85
[76]鄢丰,杨频.从竹节三七,肉桂,大黄中提取的天然活性成分的镇痛作用研究[J].中国新医药,2003,2(5)：21-22
[77]Zou K., Zhu S., Meselhy M. R. Dammarane-type saponins for Panax japonicus and their neurite out growth activity in SKN-SH cells. J nat. prod.,2002,65(9): 1288-1289
[78]李君庆.年龄及人参皂苷Rg1对大鼠大脑皮层NO释放的影响[J].药学学报,1997,32(6)：63-64
[79]张宝恒,齐治,温淑荣.竹节人参提取物对心血管的作用及对cAMP和cGMP 含量的影响[J].中心血管病杂志,1986,14(2)：100
[80]刘家兰.竹节人参提取物对实验性心肌缺血的保护作用[J].湖北民族学院学报,1999,16(1)：626
[81]Matsuda H., Samukawa K., Fukuda S1. Studies of Panax japonicus fibrinolysis. Planta medica,1989,55(1):18
[82]陈龙全,郜红利.复方竹节三七片对大鼠佐剂性关节炎治疗作用的实验研究[J].中国实验方剂学杂志,2003：(3)：25-25
[83]闵静,敖明章,刘慧明等.竹节参总皂苷对免疫器官的影响[J].湖北职业技术学院学报,2008,11(1)：109-111
[84]Hasegawa H., Matsumiya S., Uchiyama M. Inhibitory effect of some triterpenoid saponins on glucose transport in tumor cells and its application to in vitro cytotoxic and antiviral activities. Planta medica,1994,60(3):240-243
[85]张国斌.蜈蚣三七提取物抗类风湿性关节炎免疫机理的研究[D].2006
[86]孟卫星,陈龙全,袁德培.复方竹节三七对小鼠免疫功能影响的实验研究[J].恩施医专学报,1998,15(4)：123
[87]成龙全,陈朝俊.复方竹节三七片抗风湿作用的免疫调节机制研究[J].中国医药学报,2003,18(4)：247-248
[88]陈奇修.中药药理实验方法学[M].第1版.北京：人民卫生出版社,1993：317
[89]徐叔云,卞如濂,陈修.药理实验方法学[M].第3版.北京：人民卫生出版社,2002：1420-1455
[90]莫选荣罗心静类风湿性关节炎发病机制的研究进展[J].医学临床研究,2007,24：337-340
[91]李玉洁,王强状,郑晓俊.雷公藤治疗类风湿性关节炎的机制研究现状.中国药师,2004,7(8)：639-641
[92]吴柏林.雷公藤治疗类风湿性关节炎的机理探要.实用中医内科杂志,2003,17(3)：150
[93]Cibere J., Deng Z., Lin Y. Randomized double-blind Placebo-controlled study of Tripterygium wilfordii, a Chinese herbal therapy in rheumatoid arthritis. Aithritis Rheum.2000:43(1):1345
[94]Asano K., Yu Y., Kasahara T. Inhibition of murine chronic graft-versus-hos t disease by the chloroform extract of Tripterygium wilfordii Hook f. Transpl Immunol,1997, 5:315
[95]Qiu D., Kao P. N. Immunosuppressive and anti-inflammatory mechanisms of triptolide, the principal diterpenoid from the Chinese medicinal herb Tripterygium wilfordii Hook. f. DrugsR&D.,2003,4(1):1-18
[96]喻丽珍,卞慧敏.雷公藤毒副作用的研究进展.黑龙江中医药,2005(02)：52-53
[97]张季.严春临中药雷公藤甲素药理学和毒理学研究进展.内蒙古中医药,2006(6)：55-57
[98]刘琼,王晨瑶.类风湿性关节炎动物模型的研究进展[J].医学综述,2006,12(5)：313-316
[99]Andrew D. Cook, Merrill J. Rowley, Bruce D. Wines, et al. Antibodies to the Collagen-like Region of C1q and Type Ⅱ Collagen are Independent Non-cross-reactive Populations in Systemic Lupus Erythematosus and Rheumatoid Arthritis. J Autoimmu,1994,7:369-378
[100]George R. Dodge, Isabelle Pidoux, A. Robin Poole. The Degradation of Type Ⅱ Collagen in Rheumatoid Arthritis:An Immunoelectron Microscopic Study. Matrix, 1991,11:330-338
[101]Cutolo M., Accuado S. Sex hormones, HLA and the rheumatoid arthritis. J Clin Exp Rheumat,1991,9:641-642
[102]艾景录,吕爱平.性腺功能异常型肾虚痹证发生机理的实验研究[J].中国中医基础医学杂志,1995,2：142-145
[103]Ostensen M., Veiby O. P., Raiss R. Reponses of normal and rheumatoid human anticular chondrocyte cultured under various experimental conditions in agarose. Scond J Rheumatol,1991,20:172-174
[104]吕爱平,王安民,曾晓莲.益肾镯痹丸对大鼠实验性痹证影响的病理学研究[J].中医杂志,1989,29：49-51
[105]Clague R. B., Moore L. J. IgG and IgM antibody to native type Ⅱ collagen in the arthritis serum and synovial fluid. Arthritis Rheum,1984,27:1370-1376
[106]Shinmei M., Masuda K., Kikuchi T. Produtions of cytokines by chondrocytes and its role in proteoglycan degradation. J Rheumatol supl,1991,27:89-91
[107]Shuckett R., Malemud C. J. Proteoglycans synthesized by chondrocytes of human nonarthritie and osteoarthritic cartilage. Proc Soc Exp biomed,1989,110:275-279
[108]Akaogi J., Nozaki T., Satoh M., et al. Role of PGE2 and EP receptors in the pathogenesis of rheumatoid arthritis and as a novel therapeutic strategy. Endocr. Metab. Immune Disord. Drug Targets,2006,6(4):383-394
[109]Ghosh M., Tucker D. E., Burchett S. A., et al. Properties of the Group Ⅳ phospholipase A2 family. Prog. Lipid Res.,2006,45(6):487-510
[110]Niknami M., Patel M., Witting P. K., et al. Molecules in focus:Cytosolic phospholipase A2-α. Int. J. Biochem. Cell Biol.,2009,41:994-997
[111]Uozumi N., Shimizu T. Roles for cytosolic phospholipase A2α as revealed by gene-targeted mice. Prostaglandins and Other Lipid Mediators,2002,68-69:59-69
[112]Brenner T., Brocke S., Szafer F., et al. Inhibition of nitric oxide synthase for treatment of experimental autoimmune encephalomyelitis. J. Immunol.,1997, 158(6):2940-2946
[113]Miwa M., Saura R., Hirata S., et al. Induction of apoptosis in bovine articular chondrocyte by prostaglandin E2 through cAMP-dependent pathway. Osteoarthritis Cartilage,2000,8(1):17-24
[114]Pengfei Li, James A. Cook, Gary S. Gilkeson, et al, Hongkuan Fan Increased expression of beta-arrestin 1 and 2 in murine model of RA:Isoform specific regulation of inflammation. Mol Immun.,2011,49:64-74
[115]Daniel T. O., Liu H., Morrow J. D., et al. Thromboxane A2 is a mediator of cyclooxygenase-2-dependent endothelial migration and angiogenesis. Cancer Res, 1999,59(18):4574-4577
[116]Mathis S., Jala V. R., Haribabu B. Role of leukotriene B4 receptors in rheumatoid arthritis. Autoimmun. Rev.,2007,7(1):12-17
[117]Lemos H. P., Grespan R., Vieira S. M., et al. Prostaglandin mediates IL-23/IL-17-induced neutrophil migration in inflammation by inhibiting IL-12 and IFNγ production. Proc. Natl. Acad. Sci. U. S. A.,2009,106:5954-5959
[118]Andersen T. L., Del Carmen Ovejero M., Kirkegaard T., et al. A scrutiny of matrix metalloproteinases in osteoclasts:Evidence for heterogeneity and for the presence of MMPs synthesized by other cells. Bone,2004,35 (5):1107-1119
[119]Yaqin Yu, Ran Tao, Jieping Shi, et al. A genetic study of two calcium-independent cytosolic PLA2 genes in schizophrenia. Prosta Leukot Essent Fatty Acids,2005,73: 351-354
[120]Raza S. L., Cornelius L. A. Matrix metalloproteinases:Pro- and anti-angiogenic activities J. Invest. Dermatol. Symp. Proc.,2000,5:47
[121]Fallarino F., Grohmann U., Hwang K. W., et al. Modulation of tryptophan catabolism by regulatory T cells. Nat. Immunol.,2003,4(12):1206-1212
[122]Emmanouil Galanakis, Diamantis Kofteridis, Kalliopi Stratigi, et al. Intron 4 a/b polymorphism of the endothelial NOS gene is associated with both type 1 and type2 diabetes in a genetically homogeneous population. Human Immunology,2008,69: 279-283
[123]DeSilva D. R., Jones E. A., Favata M. F., et al. Inhibition of mitogen-activated protein kinase kinase blocks T cell proliferation but does not induce or prevent anergy. J Immunol,1998,160(9):4175-4181
[124]Moses M. A. The regulation of neovascularization by matrix metalloproteinases and their inhibitors. Stem Cells,1997,15(3):180-189
[125]Hayashi T., Beck L., Rossetto C., et al. Inhibition of experimental asthma by indoleamine 2,3-dioxygenase. J Clin. Invest.,2004,114(2):270-279
[126]Criado G., Simelyte E., Inglis J. J., et al. Indoleamine 2,3 dioxygenase-mediated tryptophan catabolism regulates accumulation of Thl/Thl7 cells in the joint in collagen-induced arthritis. Arthritis Rheuma,2009,60(5):1342-1351
[127]Tuyt L. M., Dokter W. H., Birkenkamp K., et al. Extracellular-regulated kinase 1/2, Jun N-terminal kinase, and c-Jun are involved in NF-κB-dependent IL-6 expression in human monocytes. J. Immunol.,1999,162:4893-4902
[128]Pages G., Guerin S., Grall D., et al. Defective thymocyte maturation in p44 MAP kinase (Erk 1) knockout mice. Science,1999,286(5443):1374-1377
[129]Szanto S., Koreny T., Mikecz K., et al. Inhibition of indoleamine 2,3-dioxygenase-mediated tryptophan catabolism accelerates collagen-induced arthritis in mice. Arthritis res. Ther.,2007,9(3):R50
[130]Schett G, Tohidast-Akrad M., Smolen J. S., et al. Activation, differential localization and regulation of the stress-activated protein kinases, extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase, in synovial tissue and cells in rheumatoid arthritis. Arthritis Rheuma,2000, 43(11):2501-2512
[131]Scherle P. A., Jones E. A., Favata M. F., et al. Inhibition of MAP kinase kinase prevents cytokine and prostaglandin E2 production in lipopolysaccharide-stimulated monocytes. J Immunol.,1998,161(10):5681-5686
[132]Brogley M. A., Cruz M., Cheung H. S. Basic calcium phosphate crystal induction of collagenase 1 and stromelysin expression is dependent on a p42/44 mitogen-activated protein kinase signal transduction pathway. J. Cell Physiol.1999,180: 215-224
[133]Benbow U., Brinckerhoff C. E. The AP-1 site and MMP gene regulation:What is all the fuss about. Matrix Biol,1997,15:519-526
[134]Shi Y., Gaestel M. In the cellular garden of forking paths:How p38 MAPKs signal for downstream assistance. Bio. Chem.,2002,383:1519-1536
[135]Udalova I. A., Mott R., Field D., et al. Quantitative prediction of NF-κB DNA-protein interactions. Proc. Nat. Aca. Sci. U. S. A.,2002,99:8167-8172
[136]Harrison L. M., Van Haaften W. C. E., Tesh V. L. Regulation of Proinflammatory Cytokine Expression by Shiga Toxin 1 and/or Lipopolysaccharides in the Human Monocytic Cell Line THP-1. Infect. Immun,2004,72:2618-2627
[137]Foster L. C., Wiesel P., Huggins G. S., et al. Role of activating protein-1 and high mobility group-Ⅰ(Y) protein in the induction of CD44 gene expression by interleukin-1β in vascular smooth muscle cells. FASEB J.,2000,14:368-378
[138]Ono K., Han J. The p38 signal transduction pathway Activation and function. Cell. Signal,2000,12:1-13
[139]Mamani-Matsuda M., Kauss T., Al-Kharrat A., et al. Therapeutic and preventive properties of quercetin in experimental arthritis correlate with decreased macrophage inflammatory mediators. Biochemi. Pharma.,2006,72:1304-1310
[140]Nataraju A., Raghavendra Gowda C. D., Rajesh R., et al. Group IIA secretory PLA2 inhibition by ursolic acid:A potent anti-inflammatory molecule. Cur. Top. Med. Chem.,2007,7:801-809
[141]Takada Y., Ichikawa H., Badmaev V., et al. Acetyl-11-keto-β-boswellic acid potentiates apoptosis, inhibits invasion, and abolishes osteoclastogenesis by suppressing NF-κB and NF-κB-regulated gene expression. J Immunol,2006,176: 3127-3140
[142]Kimmatkar N., Thawani V., Hingorani L., et al. Efficacy and tolerability of Boswellia serrata extract in treatment of osteoarthritis of knee-A randomized double blind placebo controlled trial. Phytomed,2003,10:3-7
[143]Adcocks C,. Collin P., Buttle D. J. Catechins from green tea (Camellia sinensis) inhibit bovine and human cartilage proteoglycan and type Ⅱ collagen degradation in vitro. J Nutri.,2002,132:341-346
[144]Funk J. L., Frye J. B., Oyarzo J. N., et al. Efficacy and mechanism of action of turmeric supplements in the treatment of experimental arthritis. Arthritis Rheum., 2006,54:3452-3464
[145]曾克勤,胡永红,张明敏等.雷公藤甲素对CIA大鼠T细胞胞内细胞因子表达的影响[J].北京中医药大学学报,2004,27(5)：39
[146]Yu F., Yu F., Li R., Wang R. Inhibitory effects of the Gentiana macrophylla (Gentianaceae) extract on rheumatoid arthritis of rats. J Ethnopharmaco.,2004,95: 77-81
[147]Kim S. M, Lee S. Y, Yuk D. Y, et al. Inhibition of NF-kappaB by ginsenoside Rg3 enhances the susceptibility of colon cancer cells to docetaxel. Arch. Pharmacal. Res., 2009,32:755-765
[148]Yang D. S., Liu F., Zeng F. D., et al. Effect of sinomenine on adjuvant arthritis and its mechanisms. China j Chin materia med,2005,30:1361-1363
[149]Wang Y, Jia L., Wu C.-Y. Triptolide inhibits the differentiation of Th17 cells and suppresses collagen-induced arthritis. Scand. J Immunol,2008,68:383-390
[150]Sun B., Lv L., Lu Z.-X., et al. Study on drug-guide effect of Achyranthes bidentata in Sanmiao pill in arthritic rats. Zhongguo Zhongyao Zazhi,2008,33:2946-2949
[151]Roy D. Altman The future of herbal medicine in the treatment of osteoarthritis. Osteoarthritis Cartilage,2008,16:S10-S11
[152]Zhang L., Li J., Yu S.-C., et al. Therapeutic effects and mechanisms of total flavonoids of Turpinia Arguta Seen on adjuvant arthritis in rats. J Ethnopharmacol., 2008,116:167-172
[153]Dinesh Khanna, Gautam Sethi, Kwang Seok Ahn, et al. Amita Aggarwal and Bharat B Aggarwal. Natural products as a gold mine for arthritis treatment. Curr Opin in Pharmacol,2007,7:344-351
[154]Aggarwal B. B., Bhardwaj A., Aggarwal R. S., et al. Role of resveratrol in prevention and therapy of cancer:Preclinical and clinical studies. Anticancer Res., 2004,24:2783-2840
[155]Funk J. L., Frye J. B., Oyarzo J. N., et al. Comparative effects of two gingerol-containing zingiber officinale extracts on experimental Rheumatoid arthritis. J Nat. Prod.,2009,72:403-407
[156]Yoshida N., Kanekura T., Higashi Y., et al. Bidens pilosa suppresses interleukin-1β-induced cyclooxygenase-2 expression through the inhibition of mitogen activated protein kinases phosphorylation in normal human dermal fibroblasts. J Dermatol.,2006,33:676-683
[157]Xiaoyan Shen, Cuixian Li, Huifang Zhao, et al. Inhibitory effects of a traditional Chinese herbal formula TBL-Ⅱ on type Ⅱ collagen-induced arthritis in mice. J Ethnopharmacol,2011,134(2):399-405
[158]Hwang E.-S., Park K.-K. Magnolol suppresses metastasis via inhibition of invasion, migration, and matrix metalloproteinase-2/-9 activities in PC-3 human prostate carcinoma cells. Biosci, Biotech. Biochem.,2010,74:961-967
[159]Ahmad S. F., Khan B., Bani S., et al. Amelioration of adjuvant-induced arthritis by ursolic acid through altered Th1/Th2 cytokine production. Pharmacol. Res.,2006, 53:233-240
[160]Kiani A., Rao A., Aramburu J. Manipulating immune responses with immunosuppressive agents that target NFAT. Immunity,2000,12:359-372
[161]Ramgolam V., Ang S. G., Lai Y. H., et al. Traditional Chinese medicines as immunosuppressive agents. Ann. Aca. Med. e Singapore.,2000,29:11-16
[162]Ou Y, Li W., Li X., et al. Sinomenine reduces invasion and migration ability in fibroblast-like synoviocytes cells co-cultured with activated human monocytic THP-1 cells by inhibiting the expression of MMP-2, MMP-9, CD147. Rheuma. Int., 2010(1-7)
[163]Rostoka E., Baumane L., Isajevs S., et al. Effects of kaempferol and myricetin on inducible nitric oxide synthase expression and nitric oxide production in rats. Basic Clinic. Pharmacol. Toxicol.,2010,106:461-466
[164]Tong L., Moudgil K. D. Celastrus aculeatus Merr. suppresses the induction and progression of autoimmune arthritis by modulating immune response to heat-shock protein 65. Arthritis res. Ther.,2007,9:4 (R70)
[165]Sindhu G, Ratheesh M., Shyni G. L., et al. Inhibitory effects of Cynodon dactylon L. on inflammation and oxidative stress in adjuvant treated rats. Immunopharmacol. Immunotoxicol,2009,31:647-653
[166]Yang D. S., Liu F., Zeng F. D., et al. Effect of sinomenine on adjuvant arthritis and its mechanisms. Zhongguo Zhong yao Za zhi,2005,30:1361-1363
[167]Koprowska K., Czyz M. Molecular mechanisms of parthenolide's action:Old drug with a new face. Postepy Hig. Med. Dosw (Online) 2010,64:100-114
[168]Yu S., Zheng W., Xin N., Chi Z.-H., Wang N.-Q., Nie Y.-X., Feng W.-Y., Wang Z.-Y. Curcumin prevents dopaminergic neuronal death through inhibition of the c-Jun N-terminal kinase pathway. Rejuvenation Res.2010,13:55-64
[169]Choi H. J., Park Y. R., Nepal M., et al. Inhibition of osteoclastogenic differentiation by Ikarisoside A in RAW 264.7 cells via JNK and NF-KB signaling pathways. Eur. J. Pharmacol.,2010,636:28-35
[170]Matta R., Wang X., Ge H., et al. Triptolide induces anti-inflammatory cellular responses. Am. J. Transl. Res.,2009,1:267-282
[171]Hsu YL., Chen CY, Hou MF., et al.6-dehydrogingerdione, an active constituent of dietary ginger, induces cell cycle arrest and apoptosis through reactive oxygen species/c-Jun N-terminal kinase pathways in human breast cancer cells. Mol. Nutr. Food Res.,2010,54:1307-1317
[172]Elmali N., Esenkaya I., Harma A., et al. Effect of resveratrol in experimental osteoarthritis in rabbits. Inflamma. Res.,2005,54:158-162
[173]Yang X., Liu Y, Bao Z., et al. Simultaneous determination of three active compounds in root barks of Dictamnus dasycarpus by RP-HPLC. Zhongguo Zhongyao Zazhi,2010,35:187-190
[174]Sethi G, Kwang S. A., Pandey M. K., et al. Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-KB-regulated gene products and TAK1-mediated NF-κB activation. Blood., 2007,109:2727-2735
[175]Li H., Jia Y.-F., Pan Y, et al. Effect of tripterine on collagen-induced arthritis in rats. Acta Pharmacol. Sinica.,1997,18:270-273