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.

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。

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.

Madecassoside impedes invasion of rheumatoid fibroblast-like synoviocyte from adjuvant arthritis rats via inhibition of NF-κB-mediated matrix metalloproteinase-13 expression

Fibroblast-like synoviocytes(FLS) play a pivotal role in Rheumatoid arthritis(RA) pathogenesis through aggressive migration and invasion. Madecassoside(Madec), a triterpenoid saponin present in Centella asiatica herbs, has a potent anti-inflammatory effect. In the present study, Madec exerted an obvious therapeutic effect in reversing the histological lesions in adjuvant-induced arthritis(AIA) rats. To recognize the anti-rheumatoid potentials of Madec, we further investigated whether Madec interfered with FLS invasion and metalloproteinase(MMP) expression. In cultures of primary FLS isolated from the AIA rats, Madec(10 and 30 μmol·L–1) was proven to considerably inhibit migration and invasion of FLS induced by interleukin 1β(IL-1β), but exhibiting no obvious effect on cell proliferation. Madec repressed IL-1β-triggered FLS invasion by prohibiting the expression of MMP-13. Additionally, Madec suppressed MMP-13 transcription via inhibiting the MMP-13 promoter-binding activity of NF-κB. Our results further showed that Madec down-regulated the translocation and phosphorylation of NF-κB as demonstrated by Western blotting and immunofluorescence assays. In conclusion, our results suggest that Madec exerts anti-RA activity via inhibiting the NF-κB/MMP-13 pathway.

佐剂关节炎大鼠滑膜组织HIF-1α和VEGF及CD34表达上调促进血管新生

目的观察佐剂关节炎大鼠滑膜组织缺氧诱导因子1α(HIF-1α)、血管内皮细胞生长因子(VEGF)及CD34的表达,探讨类风湿性关节炎血管新生的机制。方法 30只大鼠随机均分为正常对照组和模型对照组,模型对照组采用Freund完全佐剂建立佐剂关节炎大鼠模型。造模成功后19 d,采用反转录PCR检测滑膜组织HIF-1α、VEGF的mRNA表达,Western blot法检测HIF-1α、VEGF的蛋白表达,免疫组织化学染色检测大鼠肺组织HIF-1α、VEGF、CD34表达。结果与正常对照组比较,模型对照组关节炎症明显,滑膜组织CD34表达增加,VEGF、HIF-1αmRNA和蛋白表达显著增加;滑膜组织VEGF mRNA表达与足趾肿胀度呈正相关,HIF-1α蛋白表达与关节炎指数呈正相关,VEGF mRNA、HIF-1α蛋白表达量与CD34呈正相关。结论佐剂关节炎大鼠滑膜组织VEGF、HIF-1α、CD34过表达与滑膜血管新生有关。

TACI-Ig对佐剂性关节炎大鼠T细胞反应的调节作用

目的：研究TACI受体融合蛋白（ TACI-Ig）对T细胞介导的免疫性关节炎的调节作用,探讨其调控T细胞反应的部分机制。方法 SD大鼠右后足跖皮内注射完全弗氏佐剂建立大鼠佐剂性关节炎（ AA）模型。随机分为8组：正常组,模型组,TACI-Ig（0．7、2．1、6．3 mg/kg,皮下注射,第16-34天）治疗组,阳性对照组：重组人Ⅱ型肿瘤坏死因子受体-Fc融合蛋白（rhTNFR：Fc）（2．8 mg/kg,皮下注射,第16-34天）和甲氨蝶呤（MTX）（0．5 mg/kg,灌胃,第16-34天）,阴性对照组：免疫球蛋白G（IgG）-Fc（6．3 mg/kg,皮下注射,第16-34天）。 X线摄片观察四肢关节破坏情况；ELISA法检测外周血和滑膜组织匀浆白细胞介素17（ IL-17）的水平；流式细胞仪检测外周血和脾脏总 CD4^＋ T 细胞（ CD3^＋CD4^＋）、活化CD4^＋T细胞（ CD4^＋CD25^＋）、未致敏CD4^＋T细胞（CD4^＋CD62L^＋）和记忆 CD4^＋ T 细胞（ CD4^＋ CD44^＋）比例；免疫组化法检测滑膜组织跨膜激活剂和钙调亲环素配体相互作用分子（ TACI）、B细胞成熟抗原（ BCMA）和B细胞活化因子受体（ BAFF-R ）受体蛋白表达。结果 TACI-Ig （6．3 mg/kg）明显减轻 AA 大鼠关节软组织肿胀；TACI-Ig （0．7、2．1、6．3 mg/kg）降低外周血和滑膜组织IL-17含量,升高总CD4^＋T细胞、活化CD4^＋T细胞、记忆 CD4^＋T细胞和未致敏CD4^＋T细胞比例,下调AA大鼠滑膜组织中TACI和BCMA蛋白表达,上调BAFF-R蛋白表达。结论 TACI-Ig调节免疫性关节炎大鼠异常的T细胞反应,改善关节损伤,发挥其免疫调控作用,可能与其调节TACI、BCMA和BAFF-R受体表达有关。

电针对佐剂性关节炎大鼠下丘脑中NF-KB表达的影响

目的:观察电针对佐剂性关节炎(AA)大鼠NF-KB在下丘脑表达的影响。探讨电针的镇痛及免疫调节作用。方法:将大鼠随机分为3组,即空白组、模型组及模型加电针组。用免疫组化方法检测各组下丘脑NF-KB(P65)的表达。结果:在模型组AA大鼠下丘脑中活化的NF-KB明显增多;与空白组比较有显著性差异。电针组较模型组NF-KB的表达减少,与模型组比较有显著性差异。结论:针刺可通过抑制活化的NF-KB在下丘脑的表达,调节NF-KB活性。针刺镇痛抗炎可通过NF-KB这一环节起到关键作用。

BAFF-BAFFR及信号转导分子参与佐剂性关节炎的免疫反应及芍药苷-6′-O-苯磺酸酯的作用

目的观察佐剂性关节炎(AA)大鼠炎症不同时期脾脏B淋巴细胞刺激因子(BAFF)及其受体(BAFFR)表达变化以及芍药苷-6′-O-苯磺酸酯(CP-25)对其的影响。方法采用完全弗氏佐剂制备大鼠AA模型,随机分为正常组、AA组、CP-25(25、50、100 mg/kg)组、甲氨蝶呤(0.5 mg/kg)组、白芍总苷(50 mg/kg)组、芍药苷(50 mg/kg)组,造模后第17~31天给药。ELISA法检测外周血BAFF水平;免疫组织化学法、流式细胞术、Q-PCR和Western blot法检测CP-25对脾脏BAFFR表达的影响;Western blot法检测脾脏肿瘤坏死因子相关因子2(TRAF2)蛋白的表达。结果与正常组比较,炎症反应期(D9)、炎症初期(D16)、炎症高峰期(D23)及炎症缓解期(D30)模型大鼠血清BAFF水平和脾脏BAFFR mRNA及蛋白表达均增加。与AA组比较,CP-25(25、50、100 mg/kg)体内给药明显下调AA大鼠脾脏BAFFR mRNA及蛋白表达。体外BAFF刺激后,大鼠脾脏细胞TRAF2及BAFFR蛋白表达显著增加,CP-25(1×10^(-6)mol/L)明显降低TRAF2及BAFFR蛋白的表达。结论 CP-25发挥炎症免疫调节作用可能与其抑制AA大鼠BAFFR受体及TRAF2信号分子表达有关。

当归拈痛汤加味膏外敷对AA大鼠关节中HIF-1α、滑膜细胞凋亡的影响

目的探讨当归拈痛汤加味膏外敷在佐剂性关节炎(AA)大鼠关节中对低氧诱导因子-1α(HIF-1α)的表达及滑膜凋亡的影响。方法 40只wistar大鼠随机分为正常组、模型组、治疗组及对照组。建立AA模型,给药后18d,取踝关节行HE染色及HIF-1α免疫组化染色,流式细胞仪检测滑膜细胞的凋亡变化。分析HIF-1α、滑膜凋亡在AA中的相关性及与节炎活动指标、滑膜病理之间的关系。结果给药18d后,治疗组HE染色显示明显减轻滑膜增殖以及炎症反应,免疫组化显示治疗组HIF-1α明显表达降低,流式细胞仪检定显示治疗组滑膜细胞凋亡率明显升高。结论HIF-1α的表达与滑膜细胞凋亡率呈明显负相关,该膏剂通过抑制HIF-1α活性,从而调控一系列下游靶基因,如滑膜细胞凋亡等,此可能是其抑制滑膜增生的作用机制之一。

青蒿琥酯对佐剂性关节炎大鼠VEGF、bFGF、TNF-α、IL-1β表达的影响

目的观察青蒿琥酯对佐剂性关节炎(AA)大鼠的治疗作用及其对大鼠血清血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平和滑膜组织VEGF、bFGF表达的影响。方法选择50只Wistar大鼠造模,成模大鼠30只,随机分为模型组、青蒿琥酯组、甲氨喋呤(MTX)组,每组10只,另设正常对照组10只,排水法测定治疗前后各组大鼠的足爪容积,HE染色、光镜下观察踝关节病理变化并计算病理积分,ELISA法测定大鼠血清VEGF、bFGF水平,RTA法测定血清中TNF-α、IL-1β水平,免疫组织化学法检测滑膜组织中VEGF、bFGF的表达。结果给药后,青蒿琥酯组与MTX组均能降低大鼠的足爪肿胀率,与模型组比较差异有统计学意义(P<0.05),青蒿琥酯组与MTX组比较差异无统计学意义(P>0.05);青蒿琥酯组、MTX组光镜下踝关节病理改变,包括炎细胞浸润、滑膜增生、软骨破坏程度均较模型组减轻,青蒿琥酯组、MTX组的滑膜病理积分与模型组比较均明显下降(P<0.05),而青蒿琥酯组与MTX组比较差异无统计学意义(P>0.05);模型组血清VEGF、bFGF、IL-1β、TNF-α水平和滑膜组织VEGF、bFGF表达明显高于正常对照组(P<0.05),用药后血清VEGF、bFGF、IL-1β、TNF-α水平和滑膜组织VEGF、bFGF表达较模型组降低(P<0.05)。结论青蒿琥酯对大鼠佐剂性关节炎具有显著治疗作用,其作用机制可能与下调血管新生相关因子VEGF、bFGF、IL-1β、TNF-α表达有关。

中药熏蒸方对佐剂性关节炎大鼠IL-1Ra/IL-1β的影响

目的:研究中药熏蒸方对佐剂性关节炎(adjuvant arthritis,AA)大鼠血清、关节滑膜组织中的白细胞介素1β(IL-1β)、IL-1受体拮抗剂(IL-1Ra)的影响,并探讨其改善类风湿性关节炎(RA)的作用机制。方法:采用弗氏完全佐剂诱导AA大鼠模型,随机分为模型组、MTX组、中药熏蒸组、MTX+中药熏蒸组,分别给予中药熏蒸,MTX(1.9 mL·kg^(-1)·w^(-1))腹腔注射,中药熏蒸+MTX(1.9 mL·kg^(-1)·w^(-1))腹腔注射,另设正常大鼠为正常组,连续给药20天后处死大鼠收集血清、关节滑膜组织。观察大鼠关节肿胀度和关节炎指数的变化,酶联免疫吸附剂(ELISA)测定检测血清IL-1β、IL-1Ra的水平,反转录聚合酶链式反应(RT-PCR)法检测滑膜组织中IL-1β、IL-1Ra mRNA表达量的变化,并观察中药熏蒸方对这些指标的影响。结果:与正常组比较,模型组关节肿胀度和关节炎指数明显升高,血清、滑膜组织IL-1β表达水平明显升高、IL-1Ra表达水平下降;中药熏蒸方能抑制模型组血清、滑膜组织IL-1β的表达,上调血清、滑膜组织IL-1Ra表达量,而中药熏蒸方和MTX两药协同的效果更加明显。结论:中药熏蒸方改善AA大鼠关节炎机制可能与恢复IL-1Ra/IL-1β的平衡相关。

HIF-1α和VEGF在佐剂性关节炎大鼠膝关节滑膜的表达

目的:探讨缺氧诱导因子-1α(Hypoxia-inducible factor-1α,HIF-1α)和血管内皮生长因子(Vascular endothelial growthfactor,VEGF)在佐剂性关节炎(AA)大鼠膝关节滑膜中的表达及其与滑膜病理积分的相关性。方法:建立AA大鼠模型,于造模第28天取膝关节滑膜,常规HE染色,计算滑膜病理积分,免疫组织化学染色检测HIF-1α和VEGF在膝关节滑膜的表达。结果:AA组大鼠滑膜HIF-1α和VEGF均明显高于正常对照组(P<0.01),二者之间呈显著正相关(P<0.01),且均与滑膜病理积分呈显著正相关(P<0.01)。结论:HIF-1α和VEGF在AA大鼠膝关节滑膜增生的过程中起重要作用,二者相互影响并促进滑膜新生血管的形成。

清热凉血养阴祛风法对佐剂性关节炎大鼠外周血T淋巴细胞亚群及相关细胞因子TNF-α、IL-1β的研究

目的:探讨清热凉血养阴祛风法对佐剂性关节炎大鼠外周血T淋巴细胞亚群及相关细胞因子TNF-α、IL-1β的影响。方法:取SD大鼠制成佐剂性关节炎模型,设正常组、模型组、痹颗粒组、实验组。造模后第7天起分别予痹颗粒1.88 g/(kg·d),实验方29 g/(kg·d),正常组、模型组予生理盐水3 mL/(只·d),连续灌胃3周。腹主动脉取血,用流式细胞技术检测外周血中T淋巴细胞亚群(CD3+CD4+、CD3+CD8+、CD4+/CD8+),用酶联免疫吸附法(ELISA)检测外周血血清中细胞因子TNF-α及IL-1β。结果:实验组CD4+、CD8+T淋巴细胞,CD4+/CD8+的比值与模型组比较差异有显著性(P<0.01或P<0.05),CD4+淋巴细胞水平降低,CD8+淋巴细胞水平升高,CD4+/CD8+的比值降低。实验组细胞因子TNF-α及IL-1β水平与模型组比较差异有显著性(P<0.01),TNF-α、IL-1β水平降低。结论:清热凉血、养阴祛风法能调节T淋巴细胞紊乱和相关细胞因子水平,对大鼠佐剂性关节炎有治疗作用。

类风宁汤对大鼠佐剂性关节炎TNF-α、1L-1的影响

目的观察类风宁汤对大鼠佐剂性关节炎病理形态学及肿瘤坏死因子-a(INF-a)、白细胞介素-1(IL-1)的影响。方法将50只大鼠按照随机分组表分为正常组、模型组、类风宁低、高剂量组和阳性对照组。除正常组外,其他各组采用完全弗氏佐剂造模后,观察类风宁汤剂对大鼠佐剂性关节炎大鼠足关节病理形态改变和INF-a、IL-1的影响。结果类风宁汤能显著抑制佐剂性关节炎模型大鼠足跖滑膜、骨和软骨组织的破坏;与模型组比较,类风宁治疗组大鼠样本中INF-a、IL-1表达的灰度值(Grey)明显升高,光密度值(Density)明显降低,差异具有统计学意义(P<0.05)。结论类风宁汤对类风湿性关节炎具有一定的治疗作用,能够降低大鼠佐剂性关节炎INF-a、IL-1表达水平,减轻炎症反应,改善关节功能,对类风湿关节炎的临床治疗提供了可靠的依据。

姜黄素对佐剂性关节炎模型大鼠HIF-1α、VEGF及VEGFR表达的影响

目的 :探讨姜黄素(CUR)对佐剂性关节炎(AA)模型大鼠低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)及其受体(VEGFR)表达的影响和CUR治疗类风湿关节炎的作用机制。方法:40只健康SD大鼠随机分为正常对照组、模型组、甲氨蝶呤组和姜黄素组。造模9 d后甲氨蝶呤组和姜黄素组分别给予腹腔注射甲氨蝶呤0.5 mg/kg,3 d 1次,连续5次;姜黄素50 mg/kg-1,1 d 1次,连续14 d。第23天引颈处死大鼠,取膝关节滑膜组织,分别采用免疫组织化学染色及PCR技术检测HIF-1α、VEGF及VEGFR的表达。结果 :与正常对照组比较,模型组HIF-1α、VEGF、VEGFR表达明显增加(P<0.0l,P<0.05);与模型组比较,姜黄素组HIF-1α及VEGFR表达明显减少(P<0.01,P<0.05)。且佐剂性关节炎大鼠滑膜中HIF-1α与VEGF呈明显正相关关系(r=0.910,P<0.01)。结论:姜黄素能够抑制AA大鼠滑膜血管新生,其机制可能与其降低HIF-1α表达而下调靶基因VEGF、VEGFR的表达有关,这可能是其治疗类风湿关节炎的作用机制之一。

调督通脉针灸法对佐剂性关节炎大鼠脊神经节核因子-κB的影响

目的研究调督通脉针灸法对佐剂性关节炎(AA)大鼠脊神经核因子-κB(NF-κB)表达的影响。方法完全弗氏佐剂诱导Wistar大鼠AA,采用免疫组织化学法检测大鼠脊神经节中NF-κB活性表达。结果针灸第14天采集标本,调督通脉针灸法对AA大鼠脊神经节NF-κB(P65)的高表达活性降低优于普通针灸(P<0.05)。结论调督通脉针灸法能够抑制脊神经节NF-κB的高表达活性,为针灸治疗RA提供更多的理论依据。

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。