REGULAR EXPRESSION OF DISCOIDIN DOMAIN RECEPTOR 2 IN THE IMPROVED ADJUVANT-INDUCED ANIMAL MODEL FOR RHEUMATOID ARTHRITIS

Objective To investigate the expression of discoidin domain receptor 2 (DDR2) of fibroblast-like synovial cells in im- proved adjuvant-induced animal (AIA) model for rheumatoid arthritis (RA) and to provide evidence for DDR2’s antagonist use clinically. Methods AIA was modified by administrating 0.1 mL of complete Freund’s adjuvant (CFA, mixed with 5 mg Bacillus Calmette-Guerin vaccine/mL) into rats’ right hind paws and 0.125 mL tumor necrosis factor-α (2 U/mL) into right ankles and subpatellar fatty tissue. The expression of DDR2 in fibroblast-like synovial cells was assessed using immunohistochemistry, immunofluorescence histochemistry, and in situ hybridization methods. Levels of anti-collagen II antibody were measured using enzyme-linked immunosorbent assay. Results Given the terms mentioned above, we found a more practical rat model, apparently decreasing immunization time (average 3-5 days). DDR2 can be detected upon the 15th day of immunization; expression gradually increased with time going on, and reaching a peak 35 days after immunization before gradually decreasing. Serum anti-collagen II antibody showed similar expression patterns as DDR2, but reached peak later than DDR2, about 40 days after immunization. Conclusion Regular expression of DDR2 in animal models infers its important role in the pathological process of RA.

Gentiopicroside, originated from Gentiana macrophylla Pall, possesses antiarthritic efficacy in adjuvant-induced arthritis rats

OBJECTIVE This work aimed to investigate the anti-rheumatoid arthritic effect of gentio.picroside from Gentiana macrophylla Pall using an animal model of adjuvant induced arthritis.METH.ODS Adjuvant arthritis was induced in fifty SD male rats,which were randomly divided into five groups(n=10):control(0.5% CMC-Na) group,AIA(rats with CFA) group,dexamethasone(1 mg·kg^(-1)) group,gentiopicroside(50 mg·kg^(-1)) group,and gentiopicroside(100 mg·kg^(-1)) group.Rats were administered intragastrically with drugs or CMC-Na once a day for a period of 2 weeks.Paw swelling,arthritic index,histological changes were assessed to evaluate the anti-arthritic effect.Weight growth,spleen and thymus indexes were also investigated in.RESULTS Gentiopicroside at dose of 100 mg·kg^(-1) significantly inhibited the secondary paw swelling(P<0.05) and arthritis index(P<0.05),decreased synovial inflammatory infil.tration,synovial hyperplasia and bone erosion.Furthermore,gentiopicroside showed no immunosup.pressive adverse effects in body weight,index of spleen and thyums compared with dexamethasone administration(P<0.05,P<0.01).CONCLUSION Gentiopicroside possessed anti-arthritic efficacy in AIA rats without immunosuppressive effects.

The effects of Fumaderm~ on immunological function and cytokines in a rat model of adjuvant-induced arthritis

Objective To study the therapeutic effect of Fumaderm in Freund’s complete adjuvant-induced arthritis(AIA)in Spraque-Dawley rats.Methods Adjuvant-induced arthritis(AIA)was established by intradermal injection of 0.1 mL of Freund’s complete adjuvant(CFA)in the palmar surface of the right hindpaw and Fumaderm was delivered by oral gavage for 28 days.After CFA injection,the edema of the hindpaw was determined every two days.On 28 days after CFA injection,the lymphocyte subsets of peripheral blood and the cytokines were determined by flow cytometry,meanwhile the histopathological examination of ankle-joints of the animals was performed.Results Fumaderm had a significant therapeutic effect on AIA.The hindpaw swelling was reduced significantly in a dose-dependent manner.The ratio of peripheral blood T lymphocytes was improved obviously.Multiparameter cytokine analysis from peripheral blood CD4+ T cells showed a decrease of proinflammatory cytokines and an increase of anti-inflammatory cytokines in Fumaderm treated animals.A strongly reduced inflammatory response in the joint synovium was observed.Conclusion Fumaderm has potential anti-inflammatory effects on AIA rats.Further investigation is needed to elucidate the molecular mechanism involved in the clinical effect observed in the AIA model.

Ginsenoside Rb1 attenuates adjuvant-induced arthritis in rats through inactivation of NF-κB signaling pathway

OBJECTIVE To investigate the anti-arthritic effect and mechanism of action of ginsenoside Rb1 on adju⁃vant-induced arthritis(AIA)in rats.METHODS Male SD rats were received 0.1 mL injections of FCA(10 g·L^-1)emulsion into the right hind metatarsal foot pad for arthritis induction.After that,rats were randomly divided into six groups,namely control group,untreated group,dexamethasone(DEX,2.5 mg·kg^-1)group,low(5 mg·kg^-1),medium(10 mg·kg^-1)and high(20 mg·kg^-1)doses of ginsenoside Rb1 groups,and treated intraperitoneally at the above dosage once a day for 2 weeks.After treatment,paw swelling and arthritis indexes were evaluated,the thymus and spleen index were calculated as well.HE staining were used to observe the joint histopathology in rats.Rat ELISA kits were used to determinate the TNF-α,IL-1βand IL-6 levels.Western blotting were used to detect the related protein expression of NF-κB signaling pathway in the tissues of inflamed joints.RESULTS Rb1 significantly decreased the paw swelling and arthritis index,Compared with AIA group.HE staining results revealed that medium and high doses of Rb1 significantly reduced synovial inflammatory cell infiltration,synovial lining hyperplasia and bone destruction,compared with AIA group.Elisa results showed that Rb1 significantly decreased the TNF-α,IL-1β and IL-6 levels(P<0.05,P<0.01).Western blotting results revealed that the expression of p-IκB and p-P65 were significantly reduced in 20 mg·kg^-1 of Rb1 group,compared with AIA group(P<0.05,P<0.01).CONCIUSION Rb1 manifests therapeutic anti-inflammatory effects on rats with AIA,poten⁃tially through a mechanism of inhibiting activation of the NF-κB.

Effect of Cornus officinalis glycoside on adjuvant-induced arthritis in rats

The aim of this study was to explore the effect of Cornus officinalis glucosides (COG) on adjuvant-induced arthritis in rats and its mechanism. Seventy-two rats were divided into six groups of normal, model, Dexasone (0.125 mg/kg), high-dose COG (240 mg/kg), mid-dose COG (120 mg/kg), and low-dose COG (60 mg/kg). Rat arthritis was induced by injection of Freund's complete adjuvant in the hind paws. All treatment started from the day the arthritis was induced. The edema degree of the adjuvant injection location was determined on days 1, 3, 5, 7, 9, 11, 13, 15, 17, 20, 23 and the opposite side was observed on days 11, 13, 15, 17, 20, 23 after the injection of adjuvant. All rats were sacrificed on day 24 after the injection of adjuvant for microscopic examination of the ankle, and for the study of the immunological molecular mechanism. The results showed that the COG significantly suppressed both the primary and secondary edema, improved pathological injuries of adjuvant arthritis (AA) rat ankles, significantly suppressed the proliferation of T lymphocytes and DTH reaction. It significantly suppressed IL-1, IL-6 and TNF-αproduction from peritoneal macrophages and PGE2 in plasma. In conclusion, the Cornus officinalis glucosides (COG) is able to prevent and cure the rat adjuvant-induced arthritis, and can suppress the production of pro-inflammatory cytokine IL-1, IL-6, TNF-αand PGE2.

Human umbilical cord mesenchymal stem cells as treatment of adjuvant rheumatoid arthritis in a rat model

AIM:To investigate the effect of human umbilical cord stem cells,both mesenchymal and hematopoietic(CD34+),in the treatment of arthritis.METHODS:Mesenchymal stem cells(MSCs) and hematopoietic(CD34+) stem cells(HSC) were isolated from human umbilical cord blood obtained from the umbilical cord of healthy pregnant donors undergoing fullterm normal vaginal delivery.MSC,HSC,methotrexate(MTX) and sterile saline were injected intra-articularly into the rat hindpaw with complete freunds adjuvant(CFA) induced arthritis after the onset of disease(day 34),when arthritis had become well established(arthritis score ≥ 2).Arthritic indices were evaluated and the levels of interleukin(IL)-1,tumor necrosis factor(TNF)-α and interferon(IFN)-γ and anti-inflammatory cytokine IL-10 in serum were determined using enzyme-linked immunosorbent assay.Animals of all groups were sacrificed 34 d after beginning treatment,except positive control(PC) which was sacrificed at 10,21 and 34 d for microscopic observation of disease progression.We used hematoxylin,eosin and Masson's trichrome stains for histopathological examination of cartilage and synovium.RESULTS:The mean arthritis scores were similar in all groups at 12 and 34 d post immunization,with no statistical significant difference.Upon the injection of stem cells(hematopoietic and mesenchymal),the overall arthritis signs were significantly improved around 21 d after receiving the injection and totally disappeared at day 34 post treatment in MSC group.Mean hindpaw diameter(mm) in the MSC rats was about half that of the PC and MTX groups(P = 0.007 and P = 0.021,respectively) and 0.6 mm less than the HSC group(P = 0.047),as indicated by paw swelling.Associated with these findings,serum levels of TNF-α,IFN-γ and IL-1 decreased significantly in HSC and MSC groups compared to PC and MTX groups(P < 0.05),while the expression of IL-10 was increased.Histopathological examination with H and E stain revealed that the MTX treated group showed significant reduction of leucocytic infiltrate and hypertrophy of the synovial tissue with moderate obliteration of the joint cavity.Stem cells treated groups(both hematopoietic CD34+ and mesenchymal),showed significant reduction in leucocytic infiltrate and hypertrophy of the synovial tissue with mild obliteration of the joint cavity.With Masson's trichrome,stain sections from the PC group showed evidence of vascular edema of almost all vessels within the synovium in nearly all arthritic rats.Vacuoles were also visible in the outer vessel wall.The vessel became hemorrhagic and finally necrotic.In addition,there was extensive fibrosis completely obliterating the joint cavity.The mean color area percentage of collagen in this group was 0.324 ± 0.096,which was significantly increased when compared to the negative control group.The mean color area percentage of collagen in hematopoietic CD34+ and mesenchymal groups was 0.176 ± 0.0137 and 0.174 ± 0.0197 respectively,which showed a marked decrement compared to the PC group,denoting a mild increase in synovial tissue collagen fibers.CONCLUSION:MSC enhance the efficacy of CFAinduced arthritis treatment,most likely through the modulation of the expression of cytokines and amelioration of pathological changes in joints.

艾灸对佐剂性关节炎大鼠滑膜组织Beclin-1、LC3-Ⅱ表达的影响

目的观察艾灸对类风湿关节炎(rheumatoid arthritis,RA)大鼠炎症因子白细胞介素-2(interleukin-2,IL-2)及滑膜细胞中自噬相关因子Beclin-1、微管相关蛋白1轻链3-Ⅱ(microtubule-associated protein1 light chain 3-Ⅱ,LC3-Ⅱ)表达的影响,探索艾灸治疗RA的作用机制。方法将36只雄性SD大鼠随机分为空白组、模型组、甲氨蝶呤组、艾灸组,每组9只。采用弗氏完全佐剂造模法制备RA大鼠模型。造模成功后艾灸组予艾灸足三里、关元,每次20 min,每天1次;甲氨蝶呤组予甲氨蝶呤0.35 mg/kg灌胃,每周2次。空白组、模型组、甲氨蝶呤组每天给予艾灸组大鼠同样时长及强度的捆绑。每组均干预3周。观察大鼠一般情况,采用足趾容积测量仪检测大鼠左后肢足趾容积,ELISA法检测血清中IL-2含量,Western blot检测大鼠踝关节滑膜组织中Beclin-1、LC3-Ⅱ蛋白相对表达量。结果与模型组比较,甲氨蝶呤组及艾灸组精神一般,反应尚可,体质量恢复,较活跃,摄食、饮水尚可,足部肿胀、红肿缓解,其局部炎症及全身多发性关节炎情况均轻于模型组。与空白组比较,模型组大鼠于造模后第3、10、17、24天足趾容积明显增大(P<0.01),结合一般情况,提示模型制备成功;甲氨蝶呤组、艾灸组足趾容积于第24天增大(P<0.05)。与模型组比较,甲氨蝶呤组造模后第17、24天足趾容积降低(P<0.05,P<0.01),艾灸组第10、17、24天足趾容积明显降低(P<0.01)。干预3周后,与空白组比较,模型组血清中IL-2含量明显升高(P<0.01),滑膜组织Beclin-1、LC3-Ⅱ蛋白表达量明显上升(P<0.01);与模型组比较,甲氨蝶呤组、艾灸组血清中IL-2含量降低(P<0.05),艾灸组滑膜组织Beclin-1、LC3-Ⅱ蛋白表达量下降(P<0.05)。结论艾灸能改善RA大鼠关节肿胀,降低IL-2含量,其作用机制可能是通过调节自噬因子Beclin-1、LC3-Ⅱ蛋白表达量有关。

新疆地产全蝎对佐剂性关节炎大鼠抗炎作用研究

目的:观察地产全蝎(Scorpio)对佐剂性关节炎模型大鼠的关节肿胀及免疫、炎症因子变化。方法:建立弗氏完全佐剂模型,60只大鼠随机分为正常组、模型组、全蝎高、中、低剂量组(1.04、0.52、0.26 g·kg^(-1))、阳性组,灌胃给予相应药物干预,期间测量大鼠体质量、足跖直径,最后检测脏器指数,采用ELISA法检测血清TNF-α、IL-6、MMP-3、IL-1β、IL-10水平。结果:与正常组相比,模型组及治疗组大鼠体质量显著降低(P<0.05),与模型组相比,全蝎高中剂量组和阳性组大鼠足跖直径显著降低(P<0.05);与正常组相比,模型组大鼠的胸腺指数和脾指数显著升高(P<0.05),与模型组相比,全蝎低剂量组和阳性组的胸腺指数显著降低(P<0.05),全蝎中剂量组和阳性组的脾脏指数显著降低(P<0.05);与正常组相比,模型组大鼠血清中TNF-α、MMP-3、IL-6、IL-1β水平均显著升高(P<0.05),IL-10水平显著降低(P<0.05)。与模型组相比,高剂量组降低MMP-3显著,中剂量组降低TNF-α、MMP-3、IL-1β、IL-6水平与升高IL-10水平显著,低剂量组降低TNF-α、MMP-3、IL-6水平显著。结论:地产全蝎能够显著改善佐剂性关节炎大鼠的右后肢足肿胀程度,提高大鼠免疫力,抑制炎性反应,表明其对类风湿关节炎具有一定的治疗前景。

The pain-related behavior changes correlate with the bone damage in a rat model of rheumatoid arthritis

Objective: In view of the extensive bone damage in rheumatoid arthritis, we used a commonly utilized animal model to detect behavioral changes in pain-related and the bone damage during the early disease, and to explore the correlation between bone damage and pain-related behavioral changes. Methods: Arthritis were induced in Sprague-Dawley (SD) male rats by injecting complete Freund's adjuvant (CFA) into the tails. Pain-related behavior changes were studied using the Hargreaves, VonFrey, and acetone tests on the 0, 7, 14, day and 28 day after CFA injection. The rats were sacrificed according the same schedule. The bone damage of the right proximal tibia was studied by microCT scan and bone histological slices. Results: Animals developed soft tissue inflammation and polyarthritis on 7 days after CFA injection, and arthritic score proved obvious arthritis were established within the study period. Mechanical hyperalgesia and cold allodynia were present in the affected hind paw from the 7 day through the 28 day, but the heat hyperalgesia and the mechanical allodynia lasted a short time after CFA injection. Trabecular bone number (Tb.N), Tissue Mineral Content (TMC) and Bone Volume to Tissue Volume (BV/TV) in the proximal tibia by microCT scan were also reduced after induction, especial 14 days after CFA injection. The bone histological slices showed the trabecular bone and proteoglycan diminished, the bone damage severity scores became more severely on the 7 day after CFA injection. Using analysis of covariance, these changes had statistical significance compared with baseline. By linear regression analysis demonstrated mechanical hyperalgesia and cold allodynia correlated well with arthritic score, bone damage parameters and bone damage severity scores. Conclusion: Adjuvant-induced arthritis (AA) were observed after CFA injection and lasted within the later experimental period. Pain-related behavioral changes were observed in the early time of AA. Bone damage was also occurred with arthritis development. Pain-related behavioral change correlated well with arthritic score and bone damage parameters.

Anti-arthritis effect of berberine associated with regulating energy metabolism of macrophage through AMPK/HIF-1α pathway

OBJECTIVE To investigate berberine(BBR)attenuates arthritis in adjuvant-induced arthritic(AA)rats associated with regulating the energy metabolism and correcting the polarization of macrophages through activation of AMP-activated protein kinase(AMPK)and inhibition of hypoxia inducible factor 1α(HIF-1α).METHODS AA rats were treated with BBR(40,80,or 160 mg·kg-1)from days 15 to 29 after immunization.The histopathology of ankle joint was examined through hematoxylin-eosin(HE)staining.The concentrations of tumour necrosis factor-α(TNF-α),interleukin-6(IL-6),IL-1β,IL-2,IL-17A,interferon-gamma(IFN-γ),monocyte chemotactic protein 1(MCP-1),IL-4,IL-10,transforming growth factor-β1(TGF-β1),ATP,and lactic acid were measured by using ELISA kits.The percentage of M1 and M2 macro⁃phage cells in joint tissues were evaluated by immune-fluorescence.The expressions of p-AMPK and HIF-1αin joint of AA rats were determined according to immunohistochemistry analysis.The migration of macrophage was detected by Transwell assays.The expression of inducible nitric oxide synthase(iNOS),Arginase-1(Arg1),p-AMPK,AMPK and HIF-1αwere examined by Western blotting.The labeled macrophages were observed with laser confocal microscopy.RESULTS BBR relieved signs and symptoms of AA rats and reversed pathological changes.BBR treatment group exhibited decreases in pro-inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-2,IL-17A,IFN-γ,and MCP-1)coupled with increases anti-inflammatory cytokines(IL-4,IL-10,TGF-β1)in the serum.The number of M1 macrophage was reduced,while the number of M2 macrophage was increased in BBR group joint tissues.Moreover,BBR showed marked up-regu⁃lation the expression of p-AMPK and down-regulation the expression of HIF-1αin joint of AA rats.Next in vitro study,we found BBR up-regulated the expression of p-AMPK,Arg1(M2 marker)and down-regulated the expression of HIF-1α,iNOS(M1 marker)induced by LPS in peritoneal macrophages from normal SD rat.Furthermore,BBR treatment inhibited the migration of macrophages stimulated by LPS.The level of ATP was elevated and lactic acid was reduced in LPSinduced macrophages after treated by BBR.However,Compound C significantly attenuated the effects of BBR on acti⁃vated macrophages.CONCLUSION BBR alleviates inflammation by regulating energy metabolism and correcting the polarization of macrophage through AMPK-HIF-1αpathway.BBR might have great therapeutic value for RA.

EFFECT OF ELECTROACUPUNCTURE ON THE IMMUNOREACTIVITY OF FOCAL CUTANEOUS μ-OPIOID RECEPTOR IMMUNOREACTION-POSITIVE FIBERS IN ADJUVANT ARTHRITIC RATS

客观：学习 -opioid 的效果在煽动性的皮肤织物和 electroacupuncture (EA ) 的效果的受体(粗腐殖质) 表示在在助手的粗腐殖质积极纤维的热门免疫反应(红外) 关节炎(AA ) 老鼠。方法：48 只 SD 老鼠的一个总数被使随机化进控制(n =8 ) ，模型(n= 10 ) ， focus-side-EA (n = 10 ) ， non-acupoint-EA (n = 10 ) ，并且 healthy-side-EA (n = 10 ) 组。 AA 模型被完全的 Freund 的助手的下的注射建立( CFA ， 50 L )进左后部的 paw.EA ( 4-16 Hz ， 0.5-1.5 V )为 30 min 在焦点或健康方面和 non-acupoints 上被用于“ Huantiao ”( GB 30 )和“ Yanglingquan ”( GB 34 )。使用的 Non-acupoints 是二个地点 5 公里到健康方面上的 GB 30 和 GB 34。在焦点的真皮、下的纸巾的热门粗腐殖质红外积极的纤维与 immunohistochemical 方法被染色。疼痛的严厉被把测试弄弯的脚(anklejoint ) 检测。结果：与模型组相比，“弯曲脚的测试” 20 在 第9 和 第11 天( P0.05 )并且在 non-acupoint-EA 组在 focus-side-EA 组显著地减少了在上 第8 ， 第9 并且在 CFA ( P 0.05 )的注射以后的 11thd ，都显示双边的 GB 30 和 GB 34 和 non-acupoints 的那 EA 能减轻疼痛。从第 13 天在上，没有重要差别在 3 个 EA 组和模型组(P0.05 ) 之中在“弯曲脚的测试”被发现分数。与控制组比较，在焦点织物的粗腐殖质红外积极的神经纤维的区域价值在在在模型的焦点的粗腐殖质红外积极的神经纤维的 .The 区域价值组织的 3 个 EA 组( P0.05 )是显著地更高的， 3 个 EA 组在控制组( P0.05 )比那高重要。与模型组相比，区域在组和 healthy-side-EA 组显著地增加了的 focus-side-EA (P 0.05 ) 粗腐殖质红外积极的纤维珍视；当在 non-acupoint-EA 组和 healthy-side-EA 组的粗腐殖质红外积极的纤维的那些在 focus-side-EA 是比那显著地低的时，组( P 0.05 )，和没有重要差别在粗腐殖质红外积极的纤维( P0.05 )的区域在模型组， healthy-side-EA 组和 non-acupoint-EA 组之中被发现，在焦点方面上显示 acupoints 的 EA 的更强壮的效果。结论：GB 30 和 GB 34 的 EA 能减轻煽动性的疼痛和起来调整在在 AA 老鼠的焦点的皮纸巾的粗腐殖质红外积极的纤维的表示，施加反煽动性、止痛的效果。

大鼠佐剂性关节炎模型表现特征及评价指标

目的:描述大鼠佐剂性关节炎(Adjuvant-induced arthritis,AA)临床表现主要指标的评价方法。方法:完全弗氏佐剂免疫SD大鼠诱导AA模型,进行全身及关节炎指数评分、计算关节肿胀数、测定足爪肿胀度、进行足爪X线摄片、踝关节病理检查及评分。结果:大鼠免疫后11天(d11)出现足爪红肿,全身及关节炎指数评分和关节肿胀数增加。炎症高峰期在免疫后d19～d28。大鼠出现活动障碍,可见耳朵、鼻和尾根部"关节炎"结节。X线可见关节软组织肿胀、骨密度降低、骨侵蚀病灶以及关节间隙狭窄甚至消失。踝关节病理见滑膜组织增生,炎细胞浸润,血管翳出现等。结论:大鼠AA是兼有局部关节炎症和全身表现的类风湿性关节炎动物模型。关节炎指数、关节肿胀数、足爪肿胀度、足爪X线摄片、踝关节病理等是评价AA模型临床表现的主要指标。

类风湿关节炎动物模型与临床的关系

类风湿关节炎 (rheumatoidarthritis,RA)动物模型特别是大鼠佐剂性关节炎模型和小鼠胶原诱导性关节炎模型与人RA在发病机制、临床表现等有着相似的特点 ,是研究RA、筛选抗炎免疫药物较理想的病理模型。探讨RA动物模型的发病机制及其与临床的关系 ,对进一步了解RA的病理、生理、神经内分泌免疫等特点 ;对寻找安全有效的治疗RA的药物十分重要。该文就几种RA动物模型的建立、病理特征。

雷公藤多甙对大鼠胶原免疫性关节炎及佐剂性关节炎黏膜免疫功能影响的对比研究

目的观察雷公藤多甙对大鼠胶原免疫性关节炎(collagen-Ⅱinducedarthritis,CIA)及佐剂性关节炎(adjuvantarthritis,AA)黏膜免疫功能的影响。方法分别以Ⅱ型胶原加完全弗氏佐剂和完全弗氏佐剂单独致炎免疫大鼠造成CIA和AA模型,以雷公藤多甙治疗,观察大鼠黏膜、全身免疫指标(外周血T细胞亚群)以及关节局部炎症因子(IL-6、TNF-α、COX-2和NF-κB等)的变化。结果CIA模型组派伊尔结(Peyer’spatch,PP)CD4+、血中CD4+、CD8+均升高;AA模型组PPCD4+降低,CD8+升高,血中的CD4+,CD8+均升高。雷公藤对两模型组的PP和血中T细胞亚群存在不同影响。两种模型大鼠关节可见高水平IL-6、TNF-α、COX-2和NF-κB表达,雷公藤多甙可以抑制这些炎症介质的表达。结论两种关节炎模型在黏膜免疫反应方面及雷公藤多甙对这两种模型的影响存在异同。

实验性关节炎动物模型建立及病理机制研究进展

类风湿关节炎(rheumatoid arthritis,RA)是一种慢性系统性的免疫性炎症疾病,病因和发病机制尚未完全清楚。胶原性关节炎大鼠、小鼠模型,佐剂性关节炎大鼠模型,多聚糖蛋白诱导的关节炎小鼠模型,链球菌细胞壁诱导的关节炎大鼠、小鼠模型和转基因小鼠模型与人RA发病机制和病理变化等有着相似特点,是研究RA、筛选抗炎免疫药物较理想的实验性病理模型。探讨RA动物模型的发病机制及其与临床的关系,对进一步了解RA病理、生理、免疫等特点及寻找安全有效的治疗RA的药物十分重要。该文将对几种常用的实验性关节炎模型的建立、造模特点、发病机制、与RA的异同等方面进行综述。

祖师麻提取物的镇痛与抗炎作用研究

目的观察祖师麻提取物的抗炎、镇痛和抗佐剂性关节炎的作用。方法采用醋酸扭体法、小鼠腹腔毛细血管通透性试验、二甲苯致小鼠耳肿胀试验和弗氏完全佐剂致大鼠佐剂性关节炎试验。结果祖师麻提取物0.04、0.08g/kg可明显减少冰醋酸所致小鼠的扭体反应次数和减轻佐剂致原发性大鼠右后足跖肿胀(P<0.05、0.01);祖师麻提取物0.08g/kg能显著抑制冰醋酸致小鼠毛细血管通透性增加(P<0.05)和明显减轻小鼠耳廓肿胀度(P<0.01)。结论祖师麻提取物具有较好的镇痛、抗急性炎症和抗佐剂性关节炎的作用。

威灵仙总皂苷抑制佐剂性关节炎大鼠JAK2/STAT3信号通路

目的研究威灵仙总皂苷(TSC)对佐剂性关节炎(AA)大鼠的JAK2/STAT3信号通路的作用机制。方法雄性SD大鼠60只,随机分为6组,每组10只。除正常组外,采用Freund完全佐剂诱导AA大鼠模型。造模第12天,灌胃给予相应药物,1次/d,连续16 d,观察药物对AA大鼠体质量及足肿胀度的影响;取固定部位踝关节组织,HE染色观察病理学改变;实时荧光定量PCR法测定滑膜细胞中Janus激酶2(JAK2)、信号转导子和转录激活子3(STAT3)mRNA表达;Western blot法检测磷酸化JAK2及非磷酸化JAK2(p-JAK2、JAK2)、磷酸化STAT3及非磷酸化STAT3(p-STAT3、STAT3)的蛋白表达。结果 TSC可有效缓解AA大鼠体质量减轻及明显抑制AA大鼠足肿胀,明显改变炎细胞浸润,减少血管翳生成,减轻组织增生,有效抑制JAK2、STAT3 mRNA表达及降低p-JAK2/JAK2、p-STAT3/STAT3的相对表达。结论 TSC降低AA大鼠JAK2、STAT3 mRNA表达并抑制p-JAK2/JAK2、p-STAT3/STAT3相对表达,抑制JAK2/STAT3信号通路。

雷公藤甲素对佐剂关节炎肺功能降低大鼠Notch通路受体和配体基因表达的影响

目的观察雷公藤甲素(TPT)对佐剂关节炎(AA)大鼠Notch受体、配体表达的影响。方法将40只大鼠随机分为正常对照(NC)组、模型(MC)组、甲氨喋呤(MTX)组、TPT组,每组10只,分别向除NC组外的其余动物右后足跖皮内注射弗氏完全佐剂0.1 m L致炎,复制AA大鼠模型,致炎后第12填开始给药。NC组及MC组均给予生理盐水,其余组分别给予MTX、TPT。给药30 d后,检测各组足趾肿胀度(E)、关节炎指数(AI)、肺功能、Notch受体/配体的变化。结果 MC组大鼠E、AI、Notch3、Notch4、Delta1表达明显升高;肺功能参数、Notch1、Jagged1、Jagged2表达降低(P<0.01);与MC组比较,TPT组肺功能参数、Notch1、Jagged1、Jagged2的表达升高,E、AI及Notch3、Notch4、Delta1表达降低,疗效优于对照组MTX(P<0.05,P<0.01)。结论 TPT可能是通过上调Delta1、Notch3、Notch4的表达、下调Jagged1、Jagged2、Notch1表达,降低炎症反应和免疫复合物沉积,改善AA关节和肺部症状。

酸敏感离子通道阻断剂阿米洛利对佐剂性关节炎大鼠关节软骨细胞凋亡抑制作用的初步研究

目的:研究酸敏感离子通道阻断剂阿米洛利(Amiloride)对佐剂性关节炎(AIA)大鼠关节软骨组织的保护作用及机制。方法:将大鼠随机分为正常组、AIA模型组、阿米洛利(2.5、5.0、10.0mg·kg-1.d-1)组和阿司匹林(50mg/kg)对照组。弗氏完全佐剂(CFA)致AIA后第10天,大鼠出现继发性炎症,此时阿米洛利、阿司匹林灌胃给药,连续8d;正常组与模型组给予等容量的无菌注射用水。检测大鼠继发侧关节肿胀度,光学显微镜观察关节病理变化,透射电镜、Tunel法观察阿米洛利对AIA大鼠关节软骨细胞凋亡的影响,免疫组织化学技术测定阿米洛利对AIA大鼠关节软骨中的Ⅱ型胶原(COII)蛋白合成的影响,Alcian染色测定阿米洛利对AIA大鼠关节软骨中的PG合成的影响。结果:阿米洛利各剂量组对AIA大鼠的足肿胀未见明显影响;病理学观察关节软骨表面光滑,软骨细胞层次尚清晰,成熟软骨细胞数量增加;透射电镜观察软骨细胞核内异染色质轻度边集、细胞内粗面内质网轻度扩张、线粒体轻度肿胀,与模型组软骨细胞核膜皱缩、核染色质高度边集、核膜破裂、出现凋亡小体相比无明显细胞凋亡的特征;Tunel法原位检测显示凋亡阳性细胞数显著减少(P<0.01);免疫组化及Alcian染色检测显示关节软骨基质成分COII蛋白和PG的表达量增加。结论:ASICs阻断剂阿米洛利可通过抑制AIA大鼠关节软骨细胞凋亡发挥对关节软骨的保护作用。

独一味对大鼠佐剂性关节炎防治作用的实验研究

目的:观察独一味对大鼠佐剂性关节炎的防治作用。方法:用完全弗氏佐剂制作大鼠关节炎模型,观察独一味的影响。结果:独一味预防给药可减轻大鼠致炎侧足肿胀,也能抑制对侧足肿胀。与模型组比较,足容积差异有统计学意义(P<0.05)。致炎第19天大鼠关节炎综合评分,独一味组和模型组差异有显著统计学意义(P<0.01),独一味治疗性给药可使大鼠致炎对侧足的肿胀明显减轻。结论:独一味能明显抑制大鼠佐剂性关节炎原发病变和继发病变的形成,对继发病变有较显著的治疗效果,其减轻肿胀的作用可能与抑制炎性组织PGE2的释放有关。