重组腺病毒介导的内皮抑素抗关节炎大鼠滑膜血管新生机制的研究

目的研究重组腺病毒介导的人内皮抑素(rAD-GFP-ES)对大鼠Ⅱ型胶原诱导型关节炎(CIA)血管新生的影响。方法首先,扩增并纯化重组腺病毒。然后,建立CIA模型,自造模次日治疗组给予rAD-GFP-ES(1×1011pfu·kg-1·wk-1×4wk),在第4周取膝关节进行免疫组化检测其滑膜的微血管密度(MVD),取血清进行Western blot检测大鼠体内内皮抑素(ES)、血管内皮细胞生长因子(VEGF)、基质金属蛋白酶-3(MMP-3)的表达并计算相对含量。结果①感染rAD-GFP-ES的大鼠体内ES获得了稳定表达,是非模型组的2.4倍。②感染rAD-GFP-ES可使滑膜新生血管数目明显减少,同时能降低VEGF和MMP-3的表达。结论VEGF和MMP-3可能共同促进CIA大鼠滑膜血管新生的形成和发展,rAD-GFP-ES可能通过抑制VEGF和MMP-3的表达而发挥抗滑膜血管新生的作用。

Effect of antiangiogenesis of endostatin on rats with collagen type Ⅱ induced arthritis

Aim To investigate the effect of recombinant adenovirus mediating human endostatin (rAD-GFP-ES) on rats with collagen type Ⅱ induced arthritis (CIA), and explore the mechanism of inhibiting angiogenesis on rats CIA.Methods rAD-GFP-ES were amplified and purified. The model of rat CIA was induced by injection of intradermal collagen type Ⅱ combined with complete Freund′s adjuvant(CFA).On the second day after the injection, the therapeutic administration of rAD-GFP-ES(1×1011 pfu·kg-1·week-1×4 weeks) was performed to the rats. Knee joints from the rats sacrificed at the fourth week were estimated in morphology, and the microvessel density(MVD) of synovial membrane was detected by using immunohistochemistry assay. The relative concentration of ES, vascular endothelial growth factor(VEGF) and matrix metalloproteinase-3 (MMP-3) in sera were evaluated by Western Blot.Results ① The concentration of ES in sera of the group with the therapeutic administration of rAD-GFP-ES increased steady expression by 2.4-fold of the normal group.② The new blood vessels were less in the thin synovial tissue of rats treated with rAD-GFP-ES than that in CIA control. The administration of rAD-GFP-ES significantly decreased the production of VEGF and MMP-3 in sera.Conclusions VEGF and MMP-3 might be involved in the pathogenesis of angiogenesis of RA. rAD-GFP-ES had an inhibitory effect on the expression of VEGF and MMP-3 in rat CIA,which was related to its antiangiogenesis.