清络通痹颗粒对胶原性关节炎大鼠T细胞亚群及滑膜细胞的影响

目的：观察清络通痹颗粒(QLT)对胶原性关节炎大鼠T细胞亚群及滑膜细胞TNF-α,IL-1的影响。方法：建立胶原性关节炎(CIA)大鼠模型,同时设正常对照组．给药自d 7始,持续21 d。采用流式细胞术、酶联免疫吸附(EuSA)法、RT-PCR技术,观察QLT对CIA大鼠外周血T细胞亚群及滑膜细胞分泌TNF-α,IL-1和TNF-αmRNA,IL-1 mRNA表达的影响。结果：QLT 14．4 g·kg~(-1)可明显升高CIA大鼠外周血CD4~+T细胞,降低CD8~+T细胞,升高CD4~+/CD8~+比值(P＜0．05,P＜0．01)；QLT 7．2和14．4 g·kg~(-1)降低滑膜细胞分泌TNF-α,IL-1(P＜0．05,P＜0．01)及TNF-αmRNA,IL-1 mRNA的表达。结论：QLT可调节CIA大鼠T细胞亚群的比例,减少炎性细胞因子的产生,提示这可能是其治疗类风湿关节炎主要作用机制的一个重要方面。     

白芍总苷对胶原性关节炎大鼠滑膜细胞的作用及机制

目的研究白芍总苷(TGP)对胶原性关节炎(CIA)大鼠滑膜细胞的作用及机制。方法采用鸡II型胶原诱导大鼠CIA模型，胶原酶和胰蛋白酶消化法分离培养大鼠滑膜细胞，透射电镜观察滑膜细胞超微结构的变化，MTT法检测滑膜细胞的增殖能力，滑膜细胞培养上清液中IL-1活性的测定采用小鼠胸腺细胞增殖法，TNFα和PGE₂含量的测定采用放射免疫测定法。结果TGP能有效改善CIA大鼠滑膜细胞超微结构的变化，抑制其过度的增殖反应和产生IL-1，TNFα和PGE₂的水平。结论TGP对CIA大鼠功能亢进的滑膜细胞具有明显的抑制作用，其作用机制可能与其抑制滑膜细胞的过度增殖和分泌能力有关。     

芍药苷对胶原性关节炎大鼠滑膜细胞G蛋白偶联信号的调节作用

目的研究芍药苷(paeoniflorin,Pae)对胶原诱导性关节炎(collagen-induced arthritis,CIA)大鼠滑膜细胞G蛋白偶联信号转导的调节作用。方法用鸡Ⅱ型胶原诱导胶原性关节炎。在免疫后d16~23,胶原性关节炎大鼠灌胃给予Pae(25、50、100mg·kg-1)。通过足爪肿胀和关节炎指数评价关节炎。采用Western blot技术检测滑膜细胞中抑制性G蛋白(Gi)表达。用放免法检测滑膜细胞中cAMP水平,用激酶发光分析法测定滑膜细胞中蛋白激酶A(PKA)的活性。结果胶原性关节炎大鼠出现明显的继发性炎症反应。滑膜细胞中Gi蛋白表达增加,而cAMP水平和PKA活性明显降低。Pae抑制胶原性关节炎大鼠的炎症应答。在100mg·kg-1,Pae抑制Gi的表达。在50mg·kg-1和100mg·kg-1剂量,Pae提高cAMP水平和PKA活性。体外研究发现,Pae(2·5、12·5、62·5mg·L-1)降低滑膜细胞中Gi的表达,而提高降低的cAMP水平和PKA活性。结论胶原性关节炎大鼠滑膜细胞中G蛋白偶联的信号转导与滑膜炎病理机制密切相关;Pae对胶原性关节炎大鼠的抗炎作用是通过调节G蛋白偶联的信号转导实现的。     

白芍总苷对IL-1α诱导胶原性关节炎大鼠成纤维样滑膜细胞功能的影响

目的研究白芍总苷(TGP)对rIL-1α诱导胶原性关节炎大鼠成纤维样滑膜细胞(FLS)的影响,并探讨其部分机制。方法鸡Ⅱ型胶原(CCⅡ)诱导大鼠胶原性关节炎(CIA);MTT法检测FLS的增殖反应;放射免疫法(RIA)测定FLS产生PGE2和cAMP的水平。结果在rIL-1α(0.01、0.1、1、10μg.L-1)和1μg.L-1rIL-1α(3、6、12、24h)体外刺激下,CIA大鼠FLS产生的PGE2和cAMP水平均升高;TGP(2.5、12.5、62.5、125、250mg.L-1)体外给药可不同程度地抑制FLS的增殖反应和PGE2产生,而进一步升高cAMP水平。结论在不同浓度同一时间和同一浓度不同时间的rIL-1α体外刺激下,FLS产生PGE2和cAMP水平均升高;TGP体外作用FLS可抑制其增殖和产生PGE2,进一步升高cAMP的水平。     

红曲对胶原诱导性关节炎大鼠的抗炎作用及机制探讨

目的:观察红曲对胶原诱导性关节炎(CIA)大鼠的抗炎作用,并探讨可能的作用机制.方法:利用牛Ⅱ型胶原建立Wistar大鼠CIA模型,大鼠在首次注射Ⅱ型胶原后d 14灌胃红曲500 mg·kg-1,qd,给药至d 48处死大鼠(共给药34 d).分别测量不同时间点大鼠的后足容积,并对关节炎的肿胀程度进行评分;对大鼠的踝关节拍摄X线片,评价CIA大鼠的软骨和骨的破坏程度;同时对大鼠踝关节的滑膜进行组织病理学评分,分析单核细胞趋化蛋白(MCP)-1和调节正常T细胞表达和分泌的细胞因子(RANTES)在CIA大鼠关节滑膜的表达情况,比较不同时间点各组大鼠血清肿瘤坏死因子-α(TNF-α)的水平.结果:在首次注射Ⅱ型胶原后11-13 d,大鼠关节炎形成.红曲组大鼠的踝关节肿胀程度于首次注射Ⅱ型胶原后第4周开始下降,与CIA模型组相比有显著差异(P＜0.05),关节炎评分与CIA模型组相比显著降低(P＜0.05).在d 48,红曲组血清中炎性细胞因子TNF-α的含量、踝关节滑膜组织中MCP-1和RANTES阳性细胞均显著低于CIA模型组(P＜0.05).结论:红曲对CIA大鼠具有较强的抗炎作用,其作用机制可能与抑制踝关节滑膜组织中MCP-1和RANTES的表达有关.     

胶原性关节炎大鼠血清中炎性细胞因子的表达

目的观察白细胞介素-1(IL-1β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-4(IL-4)和白细胞介素-10(IL-10)在Ⅱ型胶原诱导性关节炎大鼠血清中的表达.方法健康雄性Wistar大鼠随机分为对照组和模型组,模型组以牛Ⅱ型胶原皮下注射诱导建立胶原性关节炎模型.采用酶联免疫吸附法(ELISA)测定上述细胞因子的含量.结果与正常对照组比较,免疫后7周关节炎大鼠血清IL-1β、TNF-α含量明显升高(P＜0.01),IL-10含量明显降低(P＜0.01),IL-4含量虽低于正常对照组,但没有统计学差异(P＞0.08).结论细胞因子网络失衡与胶原性关节炎大鼠的发病相关.     

重组人内抑素对佐剂性关节炎大鼠滑膜细胞增殖及产生细胞因子的影响

目的观察重组人内抑素对体外培养佐剂性关节炎大鼠滑膜细胞的增殖功能及产生细胞因子的影响,探讨其治疗佐剂性关节炎的作用机制。方法采用弗氏完全佐剂(CFA)诱导的佐剂性关节炎(AA)大鼠模型,用足容积法测量关节肿胀度;采用collagenasetypeⅡ消化法分离、培养滑膜细胞,MTT法检测重组人内抑素体内用药对滑膜细胞增殖的影响;放免法检测滑膜细胞上清液中IL-1β、TNF-α的含量。结果CFA致炎后d10,AA大鼠出现继发性炎症,此时皮下注射重组人内抑素(1·25,2·5,5mg·kg-1),连续7d,对照组于d10给予MTX(1·0mg·kg-1)。各剂量组能明显抑制AA大鼠的继发性足肿胀;重组人内抑素体内用药可明显减少AA大鼠滑膜细胞数量,抑制滑膜细胞的增殖,并呈剂量依赖关系;各剂量组均可明显抑制AA大鼠滑膜细胞产生过高的IL-1β和TNF-α。结论重组人内抑素抑制AA大鼠滑膜细胞过度的增殖及过高的细胞因子是其治疗AA的途径之一。     

木瓜苷通过抑制滑膜细胞功能减轻大鼠佐剂性关节炎

目的：观察木瓜苷(GCS)对大鼠佐剂性关节炎(AA)的作用,分析该作用与改善滑膜细胞的形态和功能之间的联系,方法：弗氏完全佐剂诱导大鼠AA,足容积法测量AA大鼠继发侧足容积,对关节疼痛和多发性关节炎进行评分,用11型胶原酶和胰蛋白酶消化和分离滑膜细胞,透射电镜进行滑膜细胞的超微结构观察,小鼠胸腺细胞增殖法测定滑膜细胞培养上清液中的IL—l水平,放射免疫法测定其中TNFα和PGE2含量．结果：GCS(60,120mg／kg)连续灌胃8天,可明显减轻大鼠AA的继发性足肿胀、炎性疼痛反应和多发性关节炎指数,改善滑膜细胞形态的异常并降低其高分泌的IL-1、TNFα和PGE2含量;GCS(120mg／kg)在减轻继发性炎症、疼痛和抑制滑膜细胞分泌PGE,方面明显优于阿克他利．结论：GCS具有减轻大鼠AA的作用,该作用的发挥可通过改善滑膜细胞异常的超微结构并抑制其过度分泌IL-1、TNFα和PGE2。     

京尼平苷酸对佐剂性关节炎模型大鼠滑膜细胞体外培养增殖及分泌细胞因子的影响

目的:研究京尼平苷酸(GA)对佐剂性关节炎(AA)模型大鼠滑膜细胞体外培养增殖能力和分泌细胞因子的影响。方法:用弗氏完全佐剂建立AA模型大鼠,分离其滑膜细胞进行培养,以阳性细胞数和纯度鉴定其免疫细胞;将细胞分为对照组,GA低、中、高(10-7、10-6、10-5mol·L-1)剂量组,阳性对照(甲氨蝶呤,10-6mol·L-1)组并加入相应药物处理。MTT法检测吸光度(A)考察GA对滑膜细胞增殖的影响;流式细胞术测定细胞周期;酶联免疫吸附法检测各组滑膜细胞分泌肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-10的水平。结果:2～3代滑膜细胞纯度达到试验要求;与对照组比较,GA高剂量组和阳性对照组大鼠滑膜细胞A值,S和G2/M期细胞比例,细胞外液TNF-α、IL-1β水平明显降低(P<0.05或P<0.01),G1期细胞比例、IL-10水平明显升高(P<0.05或P<0.01)。结论:GA能显著抑制AA模型大鼠滑膜细胞的增殖,阻滞细胞周期于G1期,抑制滑膜细胞分泌TNF-α和IL-1β,促进IL-10分泌。     

可溶性鸡Ⅱ型胶原对大鼠胶原性关节炎的免疫治疗作用

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

益气清痹解毒汤联合依那西普对难治性强直性脊柱炎活动期患者血清TNF-α、IL-6的影响

目的:探讨益气清痹解毒汤联合依那西普对难治性强直性脊椎炎(AS)活动期患者肿瘤坏死因子(TNF-α)、白细胞介素(IL)-6的影响。方法:将76例难治性AS活动期患者随机分为治疗组(51例,益气清痹解毒汤联合注射用依那西普和对照组(25例,注射用依那西普),检测治疗3个月前后2组临床疗效和TNF-α、IL-6水平,并进行比较。结果:治疗组和对照组有效率分别为94.12%、64.00%,2组比较有显著性差异(P<0.05);治疗后治疗组较对照组TNF-α、IL-6含量下降更显著(P<0.05)。结论:益气清痹解毒汤联合依那西普能较快改善临床症状,明显抑制难治性AS活动期患者血清TNF-α、IL-6的表达。     

塞隆骨提取物对牛Ⅱ型胶原诱导的小鼠关节炎的治疗作用及机制研究

目的了解塞隆骨水提物(SLG-B)对胶原诱导的小鼠关节炎的治疗作用及治疗机制。方法利用牛Ⅱ型胶原诱导DBA/1小鼠类风湿性关节炎模型即CIA。SLG-B口服给药观察小鼠关节炎指数的变化情况,体外培养关节炎小鼠脾细胞及巨噬细胞,ELISA法测定IL-12、IL-2、IFN-γ、TNF-α几种细胞因子。RT-PCR测定SLG-B对关节炎小鼠巨噬细胞IL-1β、IL-6、iNOS mRNA表达的影响。结果SLG-B能明显的抑制牛Ⅱ型胶原诱导关节炎的发生,能够减轻关节炎的各种症状。SLG-B在加抗原和不加抗原的情况下都能够明显的抑制关节炎小鼠脾细胞的增殖同时发现SLG-B能够抑制IL-2、IFN-γ、L-12p40、TNF-α细胞因子的产生还能够抑制小鼠腹腔巨噬细胞IL-1、IL-6及iNOS mRNA的表达。这可能是SLG-B在小鼠关节炎中表现治疗效果的分子基础。结论SLG-B对小鼠关节炎有很好的治疗效果,其作用机制主要是通过抑制巨噬细胞及脾细胞产生炎性细胞因子和抑制炎性细胞因子mRNA的表达来达到治疗类风湿性关节炎的作用。     

Emodin inhibits proinflammatory responses and inactivates histone deacetylase 1 in hypoxic rheumatoid synoviocytes

Chronic inflammation of rheumatoid arthritis (RA) is promoted by proinflammatory cytokines and closely linked to angiogenesis. In the present study, we investigated the anti-inflammatory effects of emodin (1,3,8-trihydroxy-6-methyl-anthraquinone) isolated from the root of Rheum palmatum L. in interleukin 1 beta (IL-1β) and lipopolysaccharide (LPS)-stimulated RA synoviocytes under hypoxia. Emodin significantly inhibited IL-1β and LPS-stimulated proliferation of RA synoviocytes in a dose-dependent manner under hypoxic condition. Also, enzyme linked immunosorbent assay (ELISA) revealed that emodin significantly reduced the production of pro-inflammatory cytokines [tumor necrosis factor-alpha (TNF-α), IL-6 and IL-8], mediators [prostagladin E(2) (PGE(2)), matrix metalloproteinase (MMP)-1 and MMP-13] and vascular endothelial growth factor (VEGF) as an angiogenesis biomarker in IL-1β and LPS-treated synoviocytes under hypoxia. Consistently, emodin attenuated the expression of cyclooxygenase 2 (COX-2), VEGF, hypoxia inducible factor 1 alpha (HIF-1α), MMP-1 and MMP-13 at mRNA level in IL-1β and LPS-treated synoviocytes under hypoxia. Furthermore, emodin reduced histone deacetylase (HDAC) activity as well as suppressed the expression of HDAC1, but not HDAC2 in IL-1β and LPS-treated synoviocytes under hypoxia. Overall, these findings suggest that emodin inhibits proinflammatory cytokines and VEGF productions, and HDAC1 activity in hypoxic RA synoviocytes.     

SND-117, a sinomenine bivalent alleviates type II collagen-induced arthritis in mice

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disorder that affects about 1% of the population worldwide. RA is mainly manifested by persistent synovitis and progressive joint destruction. The aim of the present study was to examine the anti-arthritis effects of SND-117, a sinomenine bivalent that is obtained from the structure modification of a clinically available anti-RA drug, sinomenine. The arthritis model (CIA) was established by immunizing DBA/1 mice with type II collagen, and the arthritis scores including inflammation, joint destruction and bone erosion were assessed after booster immunization for 3 weeks. The levels of cytokines such as IL-1β, IL-6 and TNF-α were analyzed by quantitative PCR and ELISA. The TNF-α induced NF-κB activation in fibroblast-like synovial cells (FLSCs) was analyzed by Western blot. SND-117 significantly relieved the inflammatory symptoms of collagen-induced arthritis, reduced bone erosion and joint destruction in CIA mice. The serum levels of IL-1β, IL-6 and TNF-α of CIA mice were markedly decreased by SND-117. SND-117 also strongly inhibited the phosphorylation and nuclear translocation of NF-κB p65 in FLSCs upon TNF-α stimulation. These data demonstrated that SND-117 could effectively block the pathogenesis of collagen-induced arthritis in CIA mice via inhibition of NF-κB signaling, and might provide potential clinic benefits in rheumatoid arthritis management.     

Effects of taurochenodeoxycholic acid on adjuvant arthritis in rats

Taurochenodeoxycholic acid (TCDCA) is one of the main bioactive substances of animals' bile acid. In this study, we aimed to investigate the anti-arthritic effects and potential mechanism of TCDCA on adjuvant arthritis (AA) in rats. Freund's complete adjuvant (FCA) was used to induce AA in rats. Paw swelling, index of thymus and spleen and body weight growth rate were measured, and polyarthritis index and radiologic changes were observed. The production of TNF-α, IL-1β, IL-6 and IL-10 was detected by ELISA in serum and synoviocytes. mRNA expression of TNF-α, IL-1β, IL-6 and IL-10 was determined by real-time RT-PCR in synovium tissue and synoviocytes. In both prophylactic and therapeutic treatment, TCDCA significantly suppressed paw swelling and polyarthritis index, increased the loss body weight and index of thymus and spleen, and amended radiologic changes in AA rats. The overproduction and mRNA expression of TNF-α, IL-1β and IL-6 were remarkably suppressed in serum and synovium tissue of all TCDCA-treated rats, however, IL-10 was markedly increased in prophylactic treatment. In a definite concentration ranging from 300 μg/mL to 500 μg/mL, TCDCA showed marked inhibition in the overproduction and mRNA expression of TNF-α, IL-1β and IL-6 in synoviocytes in a concentration-dependent manner, but opposite action on IL-10. In conclusion, treatment with TCDCA confers a good anti-adjuvant arthritis activity in rats, which its reparative effects could be mediated via reduction of the protein and mRNA expression of TNF-α, IL-1β and IL-6, and augment of IL-10 in rats.