Effectiveness of Huai Qi Huang Granules on Juvenile Collagen-induced Arthritis and Its Influence on Pyroptosis Pathway in Synovial Tissue

Summary:Huai Qi Huang(HQH)exerts great effects in clinic,such as anti-inflammation,immune-regulation,anti-cancer,and so on.However,the mechanism by which HQH protects juvenile idiopathic arthritis(JIA)is obscure.Thus,we explored deeply the protective mechanisms in juvenile collagen-induced arthritis(CIA)rat model.Pyroptosis is Gasdermin D(GSDMD)-dependent programmed cell death,involved in many diseases,such as sepsis.We investigated whether GSDMD-induced pyroptosis take part in mechanisms of juvenile CIA arthritis.Juvenile Wistar rats(3-4 weeks)were injected intradermally with fully emulsified bovine typeⅡcollagen and complete Freund's adjuvant to establish CIA rat models.Later,the CIA rats received oral administration of HQH(4.16 g/kg)once a day from the day 21 of modeling,with the treatment lasting for 28 days.Varieties of indicators were measured for evaluation of anti-inflammation effect of HQH,including hind paw swelling,arthritis scores,micro CT,and histopathological changes and the level of pro-inflammatory cytokines in the serum,including tumor necrosis factor alpha(TNF-α)and interleukin-18(IL-18).The expression of GSDMD and caspasein the joint synovial tissues was detected.The results demonstrated that the expression of the pyroptotic protein GSDMD and its upstream caspase-1 was significantly increased in the synovial tissues of CIA rats.The treatment of HQH ameliorated the symptoms in CIA rats,reduced levels of pro-inflammatory cytokines and hind paw swelling,down-regulated the expression of GDSMD and caspase-1.GSDMDinduced pyroptosis participated in the pathogenesis of CIA rats.The study supported that HQH can effectively improve joints inflammation of juvenile collagen-induced arthritis rats by inhibiting pyroptosis pathway in the joint synovial tissues.

Effects of Triptolide on the Expression and Activity of NF-κB in Synovium of Collagen-induced Arthritis Rats

Summary： The expression and activity of NF-kB in the synovium of collagen-induced arthritis （CIA） rats was detected in order to investigate the possible therapeutic effects of triptolide on rheumatoid arthritis （RA）. The experimental Wistar rat model of CIA was set up by intradermal injection of emulsion of bovine collagen 11 and the successful rate of setting-up models was evaluated by arthritis index （AI）. Rats were grouped randomly into three groups： normal, model and treatment group. The expression of TNF-α and IL-6 in synovial fluid was detected by ELISA, and the expression and activity of NF kB in synovium by immunohistochemistry method and by electrophoretic mobility shift assay （EMSA） respectively. As compared with normal group, the expression of TNF a and IL-6 in synovia （P〈0. 05）, and the expression and activity of NF-kB （P〈0.05） in synovium were increased in model group. There was statistical difference in above-mentioned indexes between model group and treatment group. Triptolide may play a protective role in IRA via downregulating the expression and activity of NF-kB in synovium.

Therapeutic Actions of the Chinese Herbal Formulae with Cold and Heat Properties and Their Effects on Ultrastructures of Synoviocytes in Rats of the Collagen-Induced Arthritis

The therapeutic actions of Qing Luo Yin (QLY清络饮) with heat property and Wen Luo Yin (WLY温络饮) with cold property on pain, swelling of the ankle, arthritis index and ultrastructures of synoviocytes were compared in rats of type II collagen-induced arthritis (CIA), with tripterygium glycosidorum (TG) used as control. The results indicated that both QLY and WLY could reduce pain, swelling of the ankle and the arthritis index of CIA, and QLY had better effects in reducing the swelling of the ankle and controlling the secondary pathological lesions as compared with WLY. Investigation on the ultrastructures of synoviocytes indicated that both QLY and WLY could reduce the number of Golgi apparatus, rough surface endoplasmic reticulum, dense bodies, matrix filaments and vacuoles so as to suppress the excessive secretion of synoviocytes in rats of CIA.

Prediction of Response of Collagen-induced Arthritis Rats to Methotrexate： An ~1H-NMR-based Urine Metabolomic Analysis

Over one half the patients with rheumatoid arthritis （RA） are being treated with methotrexate （MTX）. Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis （CIA） and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M 1H-nuclear magnetic resonance （1H-NMR） for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats （n=20） was different from that from the non-responsive rats （n=11）. Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis （PLS-DA） （R2=0.812, Q2=0.604）. In targeted profiling, 13 endogenous metabolites （uric acid, taurine, histidine, methionine, glycine, etc.） were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that 1H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.

苗药花蝴蝶回调CIA模型鼠血细胞变化的初步研究

目的 观察苗药花蝴蝶回调胶原诱导关节炎(CIA)模型鼠外周血细胞变化的初步作用机制。方法 40只大鼠随机分为花蝴蝶低剂量治疗组(花蝴蝶低组)、花蝴蝶高剂量治疗组(花蝴蝶高组)、甲氨蝶呤治疗组(甲氨蝶呤组)、CIA模型组(模型组)和正常组,分别按相关文献资料并结合民间用药经验灌胃给药。观察大鼠足关节肿胀度,检测大鼠外周血白细胞计数、淋巴细胞计数、单核细胞计数、淋巴细胞/单核细胞、红细胞计数、血细胞比容、血小板计数和平均血小板体积等指标,研究大鼠足关节病理切片等。结果 模型组大鼠足关节肿胀指数评分明显高于花蝴蝶低组、花蝴蝶高组、甲氨蝶呤组和正常组(P<0.01);正常组、甲氨蝶呤组、花蝴蝶高组、花蝴蝶低组足关节肿胀指数评分各组间比较差异无统计学意义(P>0.05)。与其他各组比较,模型组大鼠外周血白细胞计数、淋巴细胞计数、单核细胞计数、红细胞计数、血细胞比容、血小板计数均明显升高,而模型组大鼠外周血淋巴细胞/单核细胞、平均血小板体积均降低(P<0.01)。正常组、甲氨蝶呤组、花蝴蝶高组、花蝴蝶低组外周血相关检测指标各组间比较差异无统计学意义(P>0.05)。与其他各组比较,模型组大鼠膝关节骨髓腔淋巴细胞、单核细胞评分升高,而脂肪细胞评分降低(P<0.01)。正常组、甲氨蝶呤组、花蝴蝶高组、花蝴蝶低组大鼠膝关节骨髓腔淋巴细胞、单核细胞和脂肪细胞评分各组间比较差异无统计学意义(P>0.05)。结论 苗药花蝴蝶可能通过回调骨髓淋巴细胞、单核细胞、红细胞、血细胞比容、血小板等指标升高和平均血小板体积、淋巴细胞/单核细胞比值降低,回转CIA模型鼠外周血相应血细胞变化,最终缓解CIA模型鼠症状,这可能是苗药花蝴蝶治疗类风湿关节炎的初步作用机制之一。

Nuciferine alleviates collagen-induced arthritic in rats by inhibiting the proliferation and invasion of human arthritis-derived fibroblast-like synoviocytes and rectifying Th17/Treg imbalance

Rheumatoid arthritis(RA)is a chronic autoimmune disorder marked by persistent synovial inflammation and joint degradation,posing challenges in the development of effective treatments.Nuciferine,an alkaloid found in lotus leaf,has shown promising anti-inflammatory and anti-tumor effects,yet its efficacy in RA treatment remains unexplored.This study investigated the antiproliferative effects of nuciferine on the MH7A cell line,a human RA-derived fibroblast-like synoviocyte,revealing its ability to inhibit cell proliferation,promote apoptosis,induce apoptosis,and cause G1/S phase arrest.Additionally,nuciferine significantly reduced the migration and invasion capabilities of MH7A cells.The therapeutic potential of nuciferine was further evaluated in a collagen-induced arthritis(CIA)rat model,where it markedly alleviated joint swelling,synovial hyperplasia,cartilage injury,and inflammatory infiltration.Nuciferine also improved collagen-induced bone erosion,decreased pro-inflammatory cytokines and serum immunoglobulins(IgG,IgG1,IgG2a),and restored the balance between T helper(Th)17 and regulatory T cells in the spleen of CIA rats.These results indicate that nuciferine may offer therapeutic advantages for RA by decreasing the proliferation and invasiveness of FLS cells and correcting the Th17/Treg cell imbalance in CIA rats.

Huangqi Guizhi Wuwu Decoction suppresses inflammation and bone destruction in collagen-induced arthritis mice

Objective:Rheumatoid arthritis(RA)is a chronic inflammatory and destructive arthritis,characterized by inflammatory infiltration and bone destruction.Huangqi Guizhi Wuwu Decoction(HGWD)is traditional Chinese medicine,which has been applied in the treatment of RA in clinical.The aim of this study was to investigate the therapeutic effect of HGWD on collagen-induced arthritis(CIA)mouse model.Methods:DBA/1J female mice were used to establish the collagen-induced arthritis(CIA)model.HGWD was administered intragastrically once a day for four weeks starting on the 22nd day after the first immunization.The body weight,hind paw thickness and clinical score were measured every five days.Gait analysis,histopathological staining,enzyme-linked immunosorbent assay(ELISA),ultrasound imaging and micro-computed tomography imaging were performed to determine the effects of HGWD treatment on inflammation and bone structure in this model.Moreover,Real-time PCR and Western blot analysis were used to detect inflammatory factors mRNA and protein levels after HGWD intervention in RAW 264.7 cells.Results:HGWD attenuated symptoms of arthritis,suppressed inflammatory synovium area and the serum levels of inflammatory factors,inhibited joint space enlargement in the knee and ankle joints,reduced numbers of osteoclasts,protected bone destruction,as well as improved motor function.HGWD decreased the expression of mRNA for inflammatory factors and the protein expression levels of p-NF-rB and IL-17.Conclusion:These results suggested that HGWD suppresses inflammation,attenuates bone erosion and maintains motor function in collagen-induced arthritis mice.

铁死亡诱导剂咪唑酮埃拉斯汀(IKE)通过抑制IL-6、CCL5及CXCL9分泌缓解CIA小鼠的肺纤维化

目的 探索铁死亡诱导剂咪唑酮埃拉斯汀(IKE)对胶原蛋白诱发关节炎(CIA)小鼠肺纤维化的作用及潜在机制。方法 选取8周龄~10周龄DBA/1小鼠,按照完全Freund佐剂(CFA)与鸡Ⅱ型胶原蛋白混合乳化方法建立CIA小鼠模型,设置对照组、 CIA组、 CIA联合IKE组。每2 d进行关节评分与爪垫厚度测量,39 d后收集小鼠各脏器组织。HE染色、番红-固绿染色、甲苯胺蓝染色评价关节处组织病理改变;Masson染色评价肺部组织病理改变。免疫组织化学染色法检测肺组织α平滑肌肌动蛋白(α-SMA)、成纤维细胞活化蛋白α(FAPα)、转化生长因子β(TGF-β)及Ⅰ型胶原蛋白(Col1)、白细胞介素1(IL-1)、 IL-6、 IL-17及肿瘤坏死因子α(TNF-α)的表达水平;Olink mouse exploratory panel检测小鼠血清细胞因子IL-17α、 IL-17F、 TGF-β1、整合素亚基β6(ITG-β6)、 TNF受体超家族成员11B(TNFRSF11b)、 TNF受体超家族成员12A (TNFRSF12a)、 IL-6、 IL-1α、 IL-1β、 IL-10、 TNF-α、 CC趋化因子配体5(CCL5)、 CCL2、 CXC趋化因子配体9(CXCL9)、 CXCL1、烟酰胺腺嘌呤二核苷酸激酶(NADK)、促红细胞生成素(EPO)、集落刺激因子2(CSF2)、 TGF-α、 CCL20、 CCL3水平。结果 与CIA组相比,CIA联合IKE组治疗后关节炎症及关节损伤明显缓解;肺组织α-SMA、 FAPα、 TGF-β和Col1表达水平均呈下降趋势,表明CIA小鼠肺部胶原蛋白聚集减少,组织病变明显缓解;IL-6、 CCL5、 CXCL9、 IL-6水平显著下降表明CIA小鼠肺部炎症显著缓解。结论 IKE除缓解CIA小鼠关节炎症及关节损伤外,还可通过抑制IL-6、 CCL5及CXCL9表达缓解CIA伴发的肺纤维化。

苗药花蝴蝶回调CIA模型大鼠脂肪细胞变化的研究

目的探讨苗药花蝴蝶回调牛二型胶原蛋白诱导关节炎(CIA)模型鼠脂肪细胞变化的相关指标及其机制。方法将40只雄性SD大鼠随机分为正常组(8只)及建模组(32只),建模组大鼠建模成功后分为CIA模型组、氨甲蝶呤组及花蝴蝶高剂量组、低剂量组,各8只,根据相关文献资料结合民间用药经验分别灌胃给药(正常组:生理盐水;氨甲蝶呤组:1.04 mg/kg氨甲蝶呤,每周2次;花蝴蝶高、低组:每天7.6、3.8 mg/kg花蝴蝶药液),35 d后停止。观察各组大鼠足肿胀情况,检测血液甘油三酯(TG)、类风湿因子(RF)、免疫球蛋白A(IgA)、IgG、IgM等指标,采用酶联免疫吸附测定法检测脂联素(ADPN)、白细胞介素-10(IL-10)等细胞因子,研究足关节病理切片等。采用SPSS25.0软件对实验数据进行统计学处理。结果CIA模型组大鼠关节肿胀指数评分及RF、IgM水平均明显高于其他组,IgG水平低于花蝴蝶低组而高于其他组,差异均有统计学意义(P<0.05),TG、ADPN、IL-10水平明显低于其他组,差异均有统计学意义(P<0.05)。CIA模型组大鼠足关节骨髓腔内现低度脂肪细胞,而白细胞弥漫性增生,与其他组比较,差异均有统计学意义(P<0.05)。结论苗药花蝴蝶可能通过回调CIA模型鼠脂肪细胞的减少,保证了正常的能量代谢(如TG等)和免疫调节因子分泌(如ADPN、IL-10等),促进了免疫调节作用(如回调了RF、IgG、IgM等),缓解了足关节炎症等。

Attenuation of Collagen Induced Arthritis by Centella asiatica Methanol Fraction via Modulation of Cytokines and Oxidative Stress

Objective To investigate the anti-inflammatory, antioxidant and anti-arthritic effects of Centello asiatica methanolfraction （CAME） on collagen-induced arthritis （ClA）, an animal model of rheumatoid arthritis. Methods Arthritis was induced in female wistar rats by immunization with porcine type II collagen. The CIA rats were treated orally with CaME （50, 150, and 250 mg/kg/day） for 15 d （beginning on day 21 of the experimental period）. The clinical, histological, biochemical, and immunological parameters were assessed. Results CaME treatment （150 and 250 mg/kg） significantly attenuated the severity of CIA and reduced the synovial inflammation, cartilage erosion, and bone erosion as evident from both histological and radiographic data. The escalated plasma levels of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-12 alongwith nitric oxide in CIA rats decreased significantly on CaME treatment. The serum levels of type-Ⅱ collagen antibody were significantly lower in rats of CaME （150 and 250 mg/kg） treated group than those in the arthritic group. Furthermore, by inhibiting the above mediators, CaME also contributed towards the reversal of the disturbed antioxidant levels and peroxidative damage. Conclusion Our results clearly indicate that oral administration of CaME suppresses joint inflammation, cytokine expression as well as antioxidant imbalance, thereby contributing to an amelioration of arthritis severity in CIA rats.

程氏蠲痹汤通过抑制PI3K/AKT/mTOR通路激活减轻胶原蛋白诱导关节炎(CIA)大鼠炎症

目的探索程氏蠲痹汤(JBT)对胶原蛋白诱导关节炎(CIA)大鼠治疗的可能机制。方法将雌性SD大鼠分为正常组、CIA模型组、甲氨蝶呤(MTX)组、不同剂量JBT组、LY294002(PI3K阻断剂)组。评估JBT对实验大鼠治疗前后足趾肿胀度、关节炎指数测定的影响,HE染色检测滑膜组织病变情况,ELISA检测大鼠滑膜组织匀浆中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)水平。实时定量PCR检测磷脂酰肌醇3激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、beclin-1、P62的mRNA表达水平,Western blot法检测AKT、磷酸化的AKT(p-AKT)、mTOR、磷酸化的mTOR(p-mTOR)、PI3K、磷酸化的PI3K(p-PI3K)、P62、beclin-1、微管相关蛋白1轻链3B(LC3B)蛋白表达。结果与正常组相比,给药前1 h,其余各组大鼠足趾明显肿胀。与给药前1 h相比,高剂量JBT组、MTX组、LY294002组给药30 d后,采血前2 h足趾肿胀明显缓解。JBT能显著减轻大鼠足趾肿胀度、关节炎指数评分、滑膜组织的破坏。高剂量JBT组大鼠滑膜样本IL-1β、TNF-α水平以及PI3K、AKT、mTOR、P62的mRNA表达水平降低,p-PI3K、p-AKT、p-mTOR、P62蛋白水平明显降低,beclin-1 mRNA和蛋白表达、LC3B蛋白水平明显升高。结论JBT可能通过抑制PI3K/AKT/mTOR信号通路的激活减轻关节炎症,高剂量JBT的治疗效果与MTX、LY294002的治疗效果相当。

雷公藤多苷纳米粒的制备及其对关节炎大鼠的治疗作用

目的 制备雷公藤多苷纳米粒,探究其对胶原诱导型(collagen-induced arthritis, CIA)关节炎大鼠的治疗作用。方法 运用薄膜分散法制备雷公藤多苷纳米粒,对其进行质量评估。构建CIA模型,进行药物干预,称量大鼠体质量、测定足趾肿胀度和关节炎指数;观察大鼠脏器、膝、踝关节滑膜的病理改变;检测大鼠血清中肝肾功能水平和炎症因子表达。结果 制备的雷公藤多苷纳米粒在电镜下呈圆粒状且分布均匀,性质稳定。相较于模型组,给药组大鼠左右足趾肿胀度均明显下降(P<0.01),关节炎指数明显降低(P<0.01)。其中TG-NPs组的疗效优于TG组。与正常组相比,大鼠心脏、脾、肾、睾丸指数均明显降低(P<0.05,P<0.01)。TG-NPs组膝踝关节软骨病理损伤明显减轻,凋亡的滑膜细胞增加;与模型组相比,TG-NPs组大鼠血清中的ALT、BUN、CRE水平均明显降低(P<0.05),IL-1β、TNF-α和IL-6含量下降明显(P<0.05)。结论 TG-NPs通过诱导滑膜细胞凋亡和降低炎性细胞因子的表达对CIA有较好的治疗作用,通过静脉注射血液循环,可实现药物的缓、控释,避免口服药物造成的首过效应,减轻脏器的毒性,为治疗类风湿关节炎的新型纳米制剂的开发提供了实验依据。

C57BL/6小鼠CIA模型的建立及其监测体系的初步筛选

目的 建立稳定的C5 7BL/ 6小鼠胶原诱导关节炎 (CIA)模型 ,构建进一步进行类风湿关节炎 (RA)发病机制及治疗研究的体系。方法 取 4 0只雄性C5 7BL/ 6小鼠 ,随机分为模型组和对照组。模型组采用鸡II型胶原与完全弗氏佐剂在尾部皮内注射 ,2 1天后同样方法加强免疫。定期观察两组小鼠的关节和关节外表现以及病理学改变 ;用三重免疫荧光标记、胞内细胞因子测定流式细胞术检测外周血T细胞亚群。结果 对照组小鼠均未发病。模型组小鼠在 5 0天的发病率达 70 % ,关节炎评分在 3～ 8分 ,持续时间大约 1 0 0天 ;病理切片显示典型的炎性细胞浸润 ,滑膜增生 ,软骨及软骨下骨破坏 ;外周血Th1 /Th2比值升高。结论 应用Ⅱ型胶原能在C5 7BL/ 6小鼠上成功建立CIA模型 。

益肾蠲痹丸对CIA大鼠辅助性T细胞相关细胞因子的影响

目的:探究益肾蠲痹丸对胶原诱导关节炎大鼠模型(CIA)辅助性T细胞分泌的细胞因子作用。方法:CIA模型大鼠按随机数字表法分为正常对照组、CIA模型组、阳性药组(甲氨喋呤)和益肾蠲痹丸治疗组4组各10只。治疗结束后采用Procarta PlexTM多因子免疫检测技术定量分析各组大鼠血清中IFN-γ、IL-2、IL-4、IL-6、IL-10、IL-17A的含量。结果:益肾蠲痹丸能下调CIA大鼠机体IL-2、IL-6、IL-17A的表达,同时上调IL-10表达。讨论:益肾蠲痹丸可在一定程度上调节促炎细胞因子及抗炎细胞因子的平衡,其作用靶点可能与IL-2、IL-6、IL-17A及IL-10有关。

昆明山海棠对CIA HIF-1α的影响

目的:探讨昆明山海棠(THH)对胶原诱导性大鼠关节炎(CIA)大鼠模型中对于低氧诱导因子(HIF-1α)表达的影响。方法:建立胶原诱导性关节炎(CIA)模型,动态观察关节炎指数(AI)与HE染色关节的病理改变,免疫组化法检测HIF-1α表达。结果:THH的高中低治疗组以及地塞米松治疗组与CIA模型组和正常对照组中的HIF-1α表达有统计学意义(P<0.05)。结论:THH治疗CIA具有明显疗效,提示其作用机制与调节滑膜组织内的HIF-1α的表达有关。

原花青素对CIA大鼠足爪组织中NF-κB/p65蛋白和血清中某些促炎性细胞因子表达的影响

目的研究原花青素对Ⅱ型胶原诱导的类风湿性关节炎(CIA)大鼠足爪组织中核转录因子NF-κB/p65蛋白表达的抑制作用和血清中炎性细胞因子IL-1β、TNF-α等水平的影响。方法建立Ⅱ型胶原诱导的CIA大鼠模型,免疫组化法检测足爪组织中NF-κB/p65蛋白的表达,ELISA法检测CIA大鼠血清中IL-1β和TNF-α的含量。结果原花青素能剂量依赖性地下调NF-κB/p65蛋白的表达,降低CIA大鼠血清中TNF-α和IL-1β的水平。结论原花青素对CIA大鼠炎症的抑制作用可能与其下调NF-κB/p65蛋白表达、降低促炎性细胞因子TNF-α、IL-1β的水平有关。

补肾抗风湿方药对CIA大鼠骨组织RANKL/RANK/OPG系统的影响

目的:观察补肾抗风湿方药寒痹康汤对CIA大鼠骨组织RANKL/RANK/OPG系统的影响,揭示其抗类风湿关节炎骨质侵蚀的作用机制。方法:采用弗氏不完全佐剂乳化的牛Ⅱ型胶原蛋白诱导CIA大鼠模型,将60只Wistar大鼠随机分为正常对照组,模型对照组,甲氨蝶呤组,寒痹康汤高、中、低剂量组,以甲氨蝶呤组为对照,观察不同剂量寒痹康汤对CIA大鼠骨组织RANKL、RANK和OPG表达水平的影响。结果:与正常对照组比较,模型对照组大鼠骨组织的RANKL m RNA、RANK m RNA、RANKL蛋白、RANK蛋白的表达升高,OPG m RNA、OPG蛋白的表达下降(P<0.01);与模型对照组比较,寒痹康汤各剂量组和甲氨蝶呤组的RANKL m RNA、RANK m RNA、RANKL蛋白、RANK蛋白的表达均有不同程度的下降,OPG m RNA、OPG蛋白的表达均有不同程度的升高(P<0.01或P<0.05),其中以寒痹康汤高剂量组最明显;寒痹康汤高剂量组与甲氨蝶呤组比较,差异无统计学意义(P>0.05)。结论:寒痹康汤能抑制CIA大鼠的关节炎严重度,提高骨组织中OPG蛋白和基因表达,抑制RANKL和RANK的蛋白和基因表达,提示其具有抗类风湿关节炎骨质侵蚀的作用,机制可能与调控OPG/RANKL/RANK破骨细胞分化调节信号传导系统有关。

原花青素对CIA大鼠足爪组织中MMP-9蛋白表达的抑制作用

研究原花青素对胶原性关节炎(CIA)大鼠足爪组织中基质金属蛋白酶MMP-9表达和CIA大鼠血清中炎症因子含量的影响,探讨其治疗CIA的可能机制.用鸡Ⅱ型胶原诱导大鼠成功建立大鼠CIA模型,随机分为正常对照组、模型组、地塞米松组、原花青素(1.2mg/kg、6.0mg/kg和30mg/kg)药物处理组.各组大鼠足爪组织中MMP-9蛋白的表达用免疫组化法检测,大鼠血清中TNF-α、IL-1Β和VEGF的含量用ELISA法检测.6.0mg/kg和30mg/kg原花青素能显著下调CIA大鼠足爪组织中MMP-9蛋白的表达(p<0.01),明显降低血清TNF-α、IL-1Β和VEGF的水平(p<0.01),且呈剂量依赖性,而原花青素(1.2mg/kg)对以上指标无明显变化.原花青素(6.0mg/kg和30mg/kg)对大鼠胶原性关节炎有明显抑制作用,其机制可能与其下调胶原性关节炎大鼠足爪组织中MMP-9蛋白表达,抑制促炎性细胞因子TNF-α、IL-Β和VEGF的水平有关.

黑龙骨茎叶抗类风湿性关节炎作用研究

目的:研究苗药黑龙骨茎叶对胶原诱导性关节炎(Collagen-induced Arthritis,CIA)模型大鼠类风湿性关节炎(Rheumatoid Arthritis,RA)作用,探究黑龙骨茎叶抗RA作用的潜在开发利用度。方法:SD大鼠随机选取8只作为正常组,其余大鼠建立CIA模型。造模成功后随机分为模型组,黑龙骨茎叶高剂量组(100 mg·kg^(-1))、低剂量组(25 mg·kg^(-1)),雷公藤多苷(TG)组(10 mg·kg^(-1)),每组8只。在试验因素干预后,对各组大鼠AI评分、足趾厚度及病理形态变化进行分析比较。结果:给药前后,正常组与模型组比较有极显著差异(P<0.01),说明造模成功。给药后,大鼠左、右足趾厚度测量值显示,模型组与黑龙骨茎叶高、低剂量组相比均有显著性差异(P<0.05);AI评分显示,黑龙骨茎叶高、低剂量组与模型组相比,存在显著差异,组内比较中,黑龙骨茎叶低剂量组的消肿效果优于高剂量组(P<0.05)。病理切片结果显示,与正常组相比,模型组大鼠滑膜组织内见大量、弥漫性炎性细胞浸润;与模型组相比,黑龙骨茎叶低剂量组大鼠滑膜组织内炎性细胞浸润减少。结论:黑龙骨茎叶表现出了良好的抗RA效果,具有潜在的抗RA开发利用价值。

驼乳脂对CIA模型大鼠血清炎症因子及关节滑膜中炎症基因表达的影响

探究驼乳脂对类风湿性关节炎大鼠模型的保护作用。以牛Ⅱ蛋白和弗氏完全佐剂的混合液注射于雌性Wistar大鼠皮下,构建Ⅱ型胶原诱导型关节炎(CIA)大鼠模型,成模后随机分为空白组、模型组、扶他林组和驼乳脂组。通过为期30 d的驼乳脂涂抹,测量大鼠体质量、后足肿胀度和脏器指数,观察滑膜切片病理学变化,大鼠关节骨表面变化、检测血清中炎性细胞因子质量浓度以及滑膜组织中重要基因mRNA的表达水平。经驼乳脂涂抹后,大鼠体质量明显增加;足爪肿胀显著降低(P<0.05);关节间隙和骨侵蚀明显恢复;滑膜组织中炎性细胞减少,新生细胞增加;显著提高了抗炎因子白介素-10(IL-10)的含量(P<0.05),显著降低了促炎因子白介素-1β(IL-1β)和白介素-6(IL-6)的含量(P<0.05);同时显著降低了滑膜组织中IL-1β、核转录因子-κB的抑制因子(IκB)、IκB激酶(IKK)、蛋白激酶1(RIP1)mRNA的表达量(P<0.05)。驼乳脂可通过抑制核转录因子-κB(NF-κB)通路降低炎症因子表达,从而缓解CIA大鼠关节炎。