VEGF及其特异性受体Flt-1、Flk-1在CIA小鼠关节组织中的表达与调节

目的：研究血管内皮生长因子（VEGF）及其特异性受体Fit-1、Flk-1在Ⅱ型胶原诱导的关节炎（CIA）形成期的表达与调节，探讨VEGF在类风湿关节炎发病中的作用。方法：于DBA／1J小鼠皮下注射Ⅱ型胶原制作小鼠关节炎动物模型并进行关节炎指数评价，用ELISA法和免疫组织化学技术检测关节组织内VEGF及vWF含量以及通过RT-PCR、Southernblot技术检测关节组织内VEGF及其特异性受体Fit-1、Flk-1mRNA表达。结果：VEGF及vWF呈平行变化关系，均在关节炎发生后第四天达到最高，并与血管新生程度、关节炎严重程度呈正相关。关节组织内VEGFmRNA分别在279bp和304bp扩增片段有特异性表达，其特异性受体Fit-1、Fik-1mRNA在377bp和402bp扩增片段有特异性表达。结论：VEGF-Fit-Flk系统在关节炎形成早期起着重要作用，影响着实验诱导关节炎血管新生及病程。     

LY-294002对大鼠骨关节炎模型关节软骨中VEGF表达的影响

目的研究LY-294002对大鼠骨关节炎模型关节软骨中血管内皮生长因子(VEGF)表达的影响。方法健康雄性Wistar大鼠30只,随机分为对照组、骨性关节炎组(OA组)和LY-294002治疗组,每组10只。利用膝关节注射4%的木瓜蛋白酶溶液的方法制作骨关节炎模型。采用关节炎评分法评定各组大鼠平均关节炎指数(MAI),Western blot方法检测关节软骨中VEGF蛋白的表达变化,RT-PCR方法检测VEGF m RNA的表达变化。结果对照组关节无任何异常变化,OA模型组随着造模的时间延长,关节出现了重度红肿现象,与OA模型组比较,给药组的关节炎症反应呈现消退现象。术后8周检测关节软骨中VEGF蛋白及m RNA表达的变化,结果显示VEGF蛋白及m RNA在对照组大鼠关节软骨仅有微量表达,而在OA组大鼠关节软骨表达则显著升高(P0.01);与OA组相比,LY-294002治疗组大鼠关节软骨中VEGF蛋白及m RNA表达均显著降低(P0.01)。结论下调VEGF的表达很可是LY-294002参与骨关节炎的发病机制之一。     

艾灸对佐剂性类风湿关节炎大鼠模型VEGF表达的影响

目的探讨艾灸对佐剂性类风湿关节炎大鼠血管内皮生长因子(VEGF)表达的影响。方法 Wistar大鼠60只,随机分为对照组、模型组与艾灸组,每组20只。模型组和艾灸组的大鼠于右足趾皮下注射福氏完全佐剂(CFA)0.15ml,艾灸组在造模第2天进行艾灸干预,第7、14、21、28天检测关节炎评分及左右足趾关节肿胀度;ELISA法检测血清中VEGF的含量,免疫荧光、Western blot方法检测踝关节滑膜VEGF的表达。结果模型组与对照组相比关节炎指数评分明显升高,而艾灸组比模型组明显降低(P<0.05);模型组与对照组相比,血清及踝关节滑膜中VEGF表达水平均显著升高,而艾灸组比模型组显著降低(P<0.05)。结论艾灸能够降低关节炎指数评分,其机制可能与下调VEGF表达有关。     

抗VEGF中和抗体对Ⅱ型胶原诱导的关节炎形成的影响

目的：了解抗血管内皮生长因子（VEGF）中和抗体对Ⅱ型胶原（CoⅡ）诱导的关节炎（CIA）形成的影响。方法：于8～10wk龄的DBA／1J小鼠尾根部皮内注射鸡CoⅡ进行被动免疫，建立小鼠CIA模型，以CIA发生率、关节炎指数及关节组织的病理变化为指标，评价抗VEGF抗体对CIA形成的影响。结果：与对照组相比较．在CIA形成的早期，注射抗VEGF中和抗体能有效地抑制关节炎的产生及减轻其严重程度（P〈0．05）；而在CIA已形成后再注射抗VEGF中和抗体对关节炎的严重程度则无明显影响（P〉0．05）。结论：抗VEGF中和抗体明显抑制滑膜细胞增殖、血管新生及血管炎，因此有望成为类风湿性关节炎（RA）治疗的新型制剂。     

胶原性关节炎大鼠滑膜基因表达谱研究

cDNA微阵列（又称基因芯片）技术是一种基因表达水平检测的高通量技术,根据遗传学中心法则,利用基因互补配对原理,在同一载体上同时进行多基因检测,能同时准确而且快速地获得成千上万个基因在mRNA水平的表达信息.类风湿关节炎（Rheumatoid arthritis,RA）是一种以关节滑膜炎为主的自身免疫性疾病,致残率极高.滑膜类肿瘤样病变、软骨基质损害是其病理特点,认为是一种多因素参与,多基因改变协同作用的结果.为了获取更多基因表达信息,本研究采用含588个cDNA克隆的大鼠基因表达谱微阵列检测胶原性关节炎大鼠滑膜基因谱表达,并与正常大鼠进行比较.获取差异表达基因,为探讨类风湿关节炎病理机制提供线索.     

VEGF与endoglin在CIA大鼠滑膜组织中表达增高

目的 探讨胶原性关节炎大鼠滑膜组织中血管内皮生长因子(VEGF)及endoglin在蛋白水平和分子水平(mRNA)的表达与意义.方法 皮下给Wistar大鼠多点注射牛Ⅱ型胶原建立胶原诱导关节炎模型.观察关节肿胀和滑膜的病理学改变,用免疫组化法和RT-PCR技术检测VEGF与endoglin的蛋白及分子水平的表达.结果 ...     

条件性免疫反应对大鼠胶原性关节炎的疗效研究

目的 :探讨条件免疫反应对大鼠胶原性关节炎的治疗作用。方法 :建立大鼠胶原性关节炎模型。将模型大鼠分为 5组 :条件免疫反应 (CIR)组 ,以樟脑气味为条件刺激 ,甲氨蝶呤 (MTX) 泼尼松 (Pred)为非条件刺激 ,两者结合 7次 (7天 )后可建立条件免疫反应 ,然后每日再现条件刺激 ,每周条件刺激与非条件刺激结合 1次 ,共 4周 ;MTX Pred组 ,MTX Pred治疗 4周 ;MTX Pred减量组 ,MTX Pred治疗 7天内每天 1次 ,7天后每周 1次 ,共 4周 ;单纯闻樟脑气味组 ,单纯闻樟脑气味 4周 ;空白对照组 ,安慰剂治疗 4周。观察指标为关节炎指数评分、足掌体积、运动平衡能力、踝关节的X线改变、滑膜及肝脏病理改变。结果 :条件免疫反应组与MTX Pred组大鼠的足掌肿胀程度、滑膜炎症反应、运动功能明显改善 ,与MTX Pred减量组、单纯闻樟脑气味组相比较 ,差异显著 (均P <0 0 5 ) ,前两组之间无显著差异 ;条件免疫反应组与MTX Pred组相比 ,肝组织损害明显减轻 (P <0 0 1)。结论 :条件免疫反应可以抑制胶原性关节炎大鼠的足掌肿胀程度、抑制滑膜炎症反应、改善运动功能、减少药物用量 ,从而减轻肝脏损害等毒副作用     

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(P0.01),二者之间呈显著正相关(P0.01),且均与滑膜病理积分呈显著正相关(P0.01)。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。     

VEGF与endoglin在CIA大鼠滑膜组织中表达增高

目的探讨胶原性关节炎大鼠滑膜组织中血管内皮生长因子(VEGF)及endoglin在蛋白水平和分子水平(mRNA)的表达与意义。方法皮下给Wistar大鼠多点注射牛Ⅱ型胶原建立胶原诱导关节炎模型。观察关节肿胀和滑膜的病理学改变,用免疫组化法和RT-PCR技术检测VEGF与endoglin的蛋白及分子水平的表达。结果大部分鼠在胶原致敏后两周左右发病,主要表现为关节红肿,活动受限。VEGF与endoglin在CIA大鼠滑膜组织中表达增高(P<0.05),且两者在CIA滑膜中的蛋白表达和分子表达呈正相关(r=0.481,P<0.05)。结论 VEGF和en-doglin共同参与了类风湿性关节炎(RA)的病理过程,它们相互促进,相互影响,调控着RA的病理过程,它们是RA滑膜增生,浸润,关节破坏的关键因子。     

抗VEGF中和抗体对II型胶原诱导的关节炎形成的影响

目的:了解抗血管内皮生长因子(VEGF)中和抗体对Ⅱ型胶原(CoⅡ)诱导的关节炎(CIA)形成的影响。方法:于8~10wk龄的DBA/1J小鼠尾根部皮内注射鸡CoⅡ进行被动免疫,建立小鼠CIA模型,以CIA发生率、关节炎指数及关节组织的病理变化为指标,评价抗VEGF抗体对CIA形成的影响。结果:与对照组相比较,在CIA形成的早期,注射抗VEGF中和抗体能有效地抑制关节炎的产生及减轻其严重程度(P<0.05);而在CIA已形成后再注射抗VEGF中和抗体对关节炎的严重程度则无明显影响(P>0.05)。结论:抗VEGF中和抗体明显抑制滑膜细胞增殖、血管新生及血管炎,因此有望成为类风湿性关节炎(RA)治疗的新型制剂。     

Inhibition of acid-sensing ion channels in articular chondrocytes by amiloride attenuates articular cartilage destruction in rats with adjuvant arthritis

Objective  

The aim of this study was to examine whether drugs such as amiloride that block acid sensing ion channels (ASICs) could attenuate articular cartilage destruction in adjuvant-induced arthritis (AA).     

Comparison of equine articular cartilage thickness in various joints

Purpose: Thicknesses of fresh equine articular cartilage surfaces from the fetlock, carpal and stifle joints were measured employing a needle probe test.

Materials and methods: Eighty-seven samples used in measurement were cultivated from fetlock, carpal and stifle joints of 12 deceased within 4?h of death. After approximately three minutes of exposure to air during dissection, all cartilage samples were preserved in a saline solution to keep the articular cartilage hydrated for testing. The thickness was measured on five different spots on the same sample. The thicknesses of the fetlock, carpus and stifle were compared.

Results: The articular cartilage of the stifle was thicker than the fetlock and carpus, while the fetlock and the carpus had similar thickness values. The average thickness of the fetlock, carpal and stifle joint are 0.86, 0.87 and 2.1?mm, respectively. They were statistically compared using the Student t-test. The differences on the articular cartilage thicknesses between the fetlock and stifle, and carpus and stifle were “very highly significant” (p?相似文献   

Docosahexaenoic acid reduces inflammation and joint destruction in mice with collagen-induced arthritis

Objective

This study was designed to determine the anti-inflammatory activity of docosahexaenoic acid (DHA), alone and in combination with eicosapentaenoic acid (EPA), in a murine model of rheumatoid arthritis, collagen induced arthritis (CIA).

Methods

The CIA was induced in DBA/1OlaHsd mice by the injection of bovine type II collagen in Freunds’s complete adjuvant on days 0 and 21. Mice were fed modified diets containing DHA and/or EPA for 4 weeks prior to the initial collagen injection until study termination at day 45. The severity of CIA was assessed by measuring erythema, edema and mobility of the digits on the fore and hind paws, as well as histology. The level of serum anti-collagen antibodies was determined by ELISA. The ex vivo effects of DHA and/or EPA on splenocyte proliferation and cytokine production were evaluated by BrdU method and ELISA.

Results

Prophylactic treatment with DHA, and not DHA/EPA, significantly reduced arthritis severity and joint damage. Treatment with DHA also decreased anti-collagen (CII) antibodies in vivo, downregulated interleukin-1β, interferonγ and upregulated protective interleukin-10 ex vivo.

Conclusion

Prophylactic treatment with DHA was efficacious in a mouse model of rheumatoid arthritis and may be a useful intervention strategy against inflammatory arthritis.     

Localized deposition of amyloid in articular cartilage

The frequency, nature and tissue distribution of localized amyloid deposits in articular cartilage of young and elderly patients, with and without evidence of arthritic disease, was determined. Localized amyloid deposits in articular cartilage were not found in young patients with osteoarthritis of the hip or chondromalacia/osteoarthritis of the patella. However, in elderly patients with osteoarthritis of the hip, amyloid deposits were commonly found, although at no greater frequency than in elderly patients with no evidence of arthritis. Amyloid deposits were commonly present (in 95% of cases) in osteoarthritis of the knee joint and in the articular cartilage of all joints containing pyrophosphate deposits. Similar deposits of amyloid were also found in the articular cartilage of 40-45% of rheumatoid joints. These findings indicate that localized amyloid deposits in the articular cartilage are largely age-related and not due to specific pathological alterations affecting articular cartilage.     

富血小板血浆干预膝关节骨性关节炎模型兔关节软骨和滑膜的改变

文题释义： 富血小板血浆(PRP)：1993年Hood等首先提出富血小板血浆概念,并发现富血小板血浆含有丰富的血小板,其数目比全血中数目高3倍以上。血小板中含有大量的生长因子,如血小板衍生生长因子、转化生长因子β、类胰岛素生长因子、表皮生长因子、血管内皮生长因子等。 Ⅱ型胶原纤维(COL Ⅱ)：胶原纤维是关节软骨基质的重要组成结构之一,其中含量最多的Ⅱ型胶原纤维是构成软骨的基本框架,具有维持关节软骨的形态结构和抗张力强度的功能。基质金属蛋白酶为Ⅱ型胶原纤维的特异性降解酶,其中基质金属蛋白酶13降解Ⅱ型胶原纤维的速度是基质金属蛋白酶1的10倍。 背景：研究显示富血小板血浆具有很强的促进软骨细胞修复和增生作用。 目的：探讨富血小板血浆在骨性关节炎中对软骨细胞修复及滑膜炎症抑制的疗效。 方法：新西兰大白兔40只,于兔耳中央动脉取血后采用Hokugo等的方法制备富血小板血浆,同时检测外周血及富血小板血浆的血小板、血小板衍生生长因子、转化生长因子β和血管内皮生长因子水平。采用前交叉韧带切断法来制作动物模型后随机将兔分为实验组和对照组,实验组双侧膝关节每周注射1次0.3 mL 富血小板血浆;对照组每周注射1次0.3 mL无菌生理盐水,共注射10周。注射后第2,4,6,8,10周对兔进行大体观察及膝关节组织学观察;检测关节软骨Ⅱ型胶原蛋白及基质金属蛋白酶13水平,并进行软骨组织Mankin评分。实验方案经重庆医科大学动物实验伦理委员会批准。 结果与结论：①富血小板血浆中血小板、血小板衍生生长因子、转化生长因子β和血管内皮生长因子水平分别是正常血中的5.5,4.8,7.7和6.2倍(均P < 0.05);②注射后第6周实验组Mankin评分小于对照组(P < 0.05);③实验组第4,6,8,10周时Ⅱ型胶原蛋白水平明显高于对照组(P < 0.05);实验组第2,4,6,8,10周时基质金属蛋白酶13水平明显小于对照组(P < 0.05);④结果表明,关节腔内注射富血小板血浆能通过缓解关节滑膜炎症及延缓甚至阻断软骨细胞的损伤来抑制骨性关节炎的进展。 ORCID: 0000-0001-6301-4790(邱皓) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

结缔组织生长因子在幼年兔关节软骨全层缺损修复中的表达及意义

目的:探讨结缔组织生长因子(CTGF)在幼年兔膝关节软骨全层缺损修复过程中的表达及意义。方法:采用兔膝关节股骨关节面全层软骨缺损自我修复模型,25只幼年新西兰白兔,随机分为5组(每组5只):对照组、损伤24 h组、1周组、4周组、8周组。应用RT-PCR技术和免疫组化技术分别检测CTGF mRNA和蛋白在修复过程中的表达。结果:幼年兔关节软骨全层损伤后自我修复良好。各组检测均见CTGF mRNA表达,损伤后各组表达水平显著高于对照组(P0.05)。免疫组化显示各组软骨细胞均有CTGF蛋白表达,阳性表达主要集中在软骨的中间带及深层带。结论:幼年兔关节软骨全层缺损自我修复过程伴有显著的CTGF表达上调,CTGF可能在关节软骨修复中发挥重要作用。     

Denaturation of type II collagen in articular cartilage in experimental murine arthritis. Evidence for collagen degradation in both reversible and irreversible cartilage damage

Degradation of type II collagen is thought to be a key step in the destruction of articular cartilage in patients with rheumatoid arthritis or osteoarthritis. The aim of this study was to investigate whether type II collagen degradation is associated with cartilage destruction. Type II collagen degradation was studied in two murine arthritis models, zymosan-induced arthritis (ZIA), which develops reversible articular cartilage damage based on proteoglycan analysis, and antigen-induced arthritis (AIA), in which there is irreversible damage to the cartilage. Type II collagen degradation was assayed immunohistochemically using the COL2-3/4m antibody which recognizes denatured type II collagen, such as is produced by collagenase cleavage. In both models, degradation of type II collagen was observed in the non-calcified articular cartilage of arthritic but not of control knees. In the patella-femoral compartment, collagen denaturation started to increase on day 3 (ZIA) and day 7 (AIA) and remained high on day 14. In contrast, in the tibia-femoral compartment, type II collagen breakdown was not increased before 14 days in either model. By 28 days, collagen denaturation was strongly reduced in the patella-femoral compartment in the ZIA model, but persisted in the tibia-femoral compartment in both models. In conclusion, increased type II collagen degradation was found in articular cartilage of both ZIA and AIA animals. Since ZIA does not develop irreversible cartilage destruction, this indicates that cartilage may have the ability to withstand a limited degree of type II collagen degradation without developing irreversible damage. Copyright © 1999 John Wiley & Sons, Ltd.     

Lectin-binding in normal and fibrillated articular cartilage of human patellae

Summary Fluorescein-isothiocyanate (FITC) labeled lectins were used to study the distribution of specific binding-sites in histological sections of normal and fibrillated articular cartilage of human patellae.It has been shown that normal articular cartilage reveals lectin binding-sites for Concanavalin A (Con A) and wheat germ agglutinin (WGA), but not for soybean agglutinin (SBA), peanut agglutinin (PNA) and Ulex europaeus agglutinin (UEA).In fibrillated cartilage the distribution pattern of Con A and WGA is completely changed. SBA, PNA and UEA show a distinct staining pattern in particular in the fibrillated areas of degenerated cartilage. Lectin-staining of the extracellular matrix and the chondrocytes in both normal and fibrillated cartilage did not show any correlation with material that was either PAS- or Alcian blue-positive. In comparison with the conventional PAS- and Alcian blue reaction lectin-staining proved to be superior.Visualization of intra- and extracellular glycoconjugate-changes in normal and fibrillated cartilage in areas with no PAS and/or Alcian blue staining indicates that all layers of the cartilage are involved in the pathological process.It is evident that lectins can demonstrate minute differences between normal and arthrotic cartilage and we therefore conclude that lectins are sensitive and specific tools for the study of degenerative joint diseases.     

Patterns of radiocarpal joint articular cartilage wear in cadavers

The radiocarpal joint transmits about 80% of the compression forces crossing the wrist. However, primary osteoarthritis of this joint is surprisingly uncommon, suggesting that articular cartilage wear is not sufficient to produce arthritic symptoms. By examining the distal radius, scaphoid, and lunate in aged cadavers, wear patterns were charted and measured, allowing assessment of radiocarpal joint wear and mechanics. Bilateral radiocarpal joints of 16 females and 14 males (age 77.7 ± 14.4, N = 30) were exposed and measurements of the wear recorded microscopically. Wear locations were mapped, and X-Y loci and wear areas calculated. Gender right and sides compared. Over 95% of distal radius wear showed distinct radial-scaphoid and radial-lunate wear areas. These bilateral areas were in the palmar half of the distal radius. One main central wear area was seen in 95% of the scaphoid, and 97% of the lunate articular surfaces that were examined. Articular wear showed a circular pattern and was minimal in 95.7% of the surfaces, and the lunate showed the largest wear area. Wear patterns in males and females support the literature that for most ADLs the wrist is in slight extension and ulnar deviation. There are gender differences, but wear areas between sides were similar. Female wear indicates their wrist is positioned more often in a more extended and ulnarly deviated position than males. The wear patterns suggest rotational movements of the scaphoid and lunate during wrist motion and that the wrist is most often used in neutral flexion/extension to slight extension.     

Amelioration of established murine collagen-induced arthritis with anti-IL-1 treatment.

Inflammatory cytokines have been implicated in the pathogenesis of rheumatoid arthritis. To validate a key role for IL-1 in arthritic processes we have studied the protective effect of neutralizing antimurine IL-1 antibodies in the murine collagen-induced arthritis (CIA) model. Combination of anti-IL-1 alpha and anti-IL-1 beta given before onset of arthritis was shown to prevent disease completely. Remarkably, a single treatment was also highly effective in the established phase of arthritis, reducing both inflammation as well as cartilage destruction. Suppression was most pronounced with the combination, but anti-IL-1 beta alone also induced significant relief. Finally, we studied the protective effect of IL-1 neutralization on cartilage metabolism in a unilateral expression model of collagen arthritis. To this end zymosan was injected in one knee joint before onset of disease, resulting in accelerated expression in that particular joint and the draining paw. Anti-IL-1 treatment started after accelerated expression of arthritis was able to fully normalize chondrocyte synthetic function, which was highly suppressed in the control group. It is concluded that IL-1 is an important determinant in both inflammation and cartilage destruction in collagen arthritis, and this may have implications for therapy in human arthritis.