Prophylactic adenovirus-mediated human kallistatin gene therapy suppresses rat arthritis by inhibiting angiogenesis and inflammation
Prophylactic adenovirus-mediated human kallistatin gene therapy suppresses rat arthritis by inhibiting angiogenesis and inflammation
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DOI：
10.1002/art.20991
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发表时间：
2005-04-01
影响因子：
--
通讯作者：
Chao, J
中科院分区：
文献类型：
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作者：
Wang, CR;Chen, SY;Chao, J
目标。卡利斯汀已被证明是一种血管生成抑制剂。在这项研究中，我们研究腺病毒介导的kallistatin基因传递是否在大鼠关节炎模型中具有预防作用。在大鼠关节炎发生之前，将含有人kalistatin基因的腺病毒(AdHKBP)注射到大鼠的踝关节内。检测Kalistatin基因转移对关节提取液中内皮细胞增殖的影响。根据临床参数，包括踝围、关节指数和放射学评分来确定对Kalistatin治疗的反应。苏木精-伊红染色，对关节组织进行评分和中性粒细胞计数。此外，通过鉴定血管性假血友病因子阳性的内皮细胞来量化小血管。通过检测踝关节匀浆中肿瘤坏死因子α(TNFα)和白介素1β(IL-1β)的水平来确定炎症反应。免疫组织化学分析和Western blotting检测基因转移后重组人kalistatin在大鼠踝关节中的表达。与携带绿色荧光蛋白的腺病毒载体组相比，AdHKBP治疗组大鼠的踝围、关节指数和放射学评分显著减少。Kalistatin基因转移也显著改善了踝关节的组织学评分，减少了血管密度和中性粒细胞数量。Kalistatin对踝关节炎症细胞聚集的抑制作用伴随着关节匀浆中肿瘤坏死因子α和IL-1β水平的降低。此外，体外实验表明，AdHKBP处理的关节提取物可明显抑制血管内皮细胞的增殖，支持Kalistatin抑制血管生成的作用。本研究证明Kalistatin基因治疗对大鼠踝关节关节炎有预防作用。卡利斯汀通过其抗血管生成和抗炎活性抑制关节炎。这些结果暗示了kallistatin基因治疗在抑制关节炎方面的潜在治疗应用。
Objective. Kallistatin has been shown to be an angiogenesis inhibitor. In this study, we investigated whether adenovirus-mediated kallistatin gene delivery has a prophylactic effect in a rat arthritis model.Methods. Adenovirus containing the human kallistatin gene (AdHKBP) was injected intraarticularly into ankle joints before the onset of arthritis in a rat model. The effect of kallistatin gene transfer on endothelial cell proliferation in joint extracts was assayed. The response to kallistatin treatment was determined according to clinical parameters, including ankle circumference, articular index, and radiographic scores. Hematoxylin and eosin staining was performed in order to score joint tissues and count neutrophil numbers. In addition, small vessels were quantified by identification of von Willebrand factor-positive endothelial cells. The inflammatory responses were determined by measuring tumor necrosis factor alpha (TNF alpha) and interleukin-1 beta (IL-1 beta) levels in ankle homogenates.Results. The expression of recombinant human kallistatin in rat ankle joints after gene transfer was identified by immunohistochemical analysis and Western blotting. Significant reductions in the ankle circumference, articular index, and radiographic score were observed in AdHKBP-treated rats compared with control rats treated with the adenoviral plasmid carrying green fluorescent protein. Kallistatin gene transfer also significantly ameliorated the histologic scores in ankle joints and reduced vessel density and neutrophil numbers. The inhibitory effect of kallistatin on the accumulation of inflammatory cells in ankle joints was accompanied by reduced TNF alpha and IL-1 beta levels in joint homogenates. Furthermore, an in vitro experiment showed that the proliferation of endothelial cells was markedly inhibited by the addition of AdHKBP-treated joint extract to the culture media, supporting a role of kallistatin in inhibiting angiogenesis.Conclusion. This study demonstrates that kallistatin gene therapy has a prophylactic effect in inhibiting arthritis in the rat ankle. Kallistatin inhibits arthritis through its antiangiogenesis and antiinflammation activities. These results implicate potential therapeutic applications for suppression of arthritis by kallistatin gene therapy.