Interleukin-1 receptor antagonist delivered directly and by gene therapy inhibits matrix degradation in the intact degenerate human intervertebral disc: an in situ zymographic and gene therapy study.
Interleukin-1 receptor antagonist delivered directly and by gene therapy inhibits matrix degradation in the intact degenerate human intervertebral disc: an in situ zymographic and gene therapy study.
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DOI：
10.1186/ar2282
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发表时间：
2007
影响因子：
4.9
通讯作者：
Freemont, Anthony J
中科院分区：
文献类型：
--
作者：
Le Maitre, Christine L;Hoyland, Judith A;Freemont, Anthony J
数据暗示IL-1在改变的基质生物学中是人类椎间盘（IVD）退变的特征。在目前的研究中，我们研究了IL-1诱导基质降解的酶机制，以及IL-1抑制剂IL-1受体拮抗剂（IL-1 Ra）是否会抑制降解。原位酶谱（ISZ）和免疫组织化学的组合被用来检查IL-1和IL-1 Ra对基质降解和金属依赖性蛋白酶（MDP）的表达在非退化和退化的人IVD的外植体的影响。ISZ采用三种底物（明胶、胶原、酪蛋白）和不同的挑战（IL-1β、IL-1 Ra和酶抑制剂）。对MDP进行免疫组织化学。此外，使用基因工程构建体将IL-1 Ra引入退化的IVD外植体中。这项研究的新发现是：直接将IL-1 Ra递送到退化IVD的外植体上消除了由多底物ISZ评估的基质降解;由ISZ评估的基质降解与由免疫组织化学定义的MDP表达之间存在直接关系;单次注射经工程改造以过表达IL-1 Ra的IVD细胞显著抑制MDP表达两周。我们的研究结果表明，IL-1是一个关键的细胞因子驱动基质降解退化的IVD。此外，直接或通过基因治疗递送的IL-1 Ra抑制IVD基质降解。IL-1 Ra可用于治疗以抑制IVD的变性。
Data implicate IL-1 in the altered matrix biology that characterizes human intervertebral disc (IVD) degeneration. In the current study we investigated the enzymic mechanism by which IL-1 induces matrix degradation in degeneration of the human IVD, and whether the IL-1 inhibitor IL-1 receptor antagonist (IL-1Ra) will inhibit degradation. A combination of in situ zymography (ISZ) and immunohistochemistry was used to examine the effects of IL-1 and IL-1Ra on matrix degradation and metal-dependent protease (MDP) expression in explants of non-degenerate and degenerate human IVDs. ISZ employed three substrates (gelatin, collagen, casein) and different challenges (IL-1β, IL-1Ra and enzyme inhibitors). Immunohistochemistry was undertaken for MDPs. In addition, IL-1Ra was introduced into degenerate IVD explants using genetically engineered constructs. The novel findings from this study are: IL-1Ra delivered directly onto explants of degenerate IVDs eliminates matrix degradation as assessed by multi-substrate ISZ; there is a direct relationship between matrix degradation assessed by ISZ and MDP expression defined by immunohistochemistry; single injections of IVD cells engineered to over-express IL-1Ra significantly inhibit MDP expression for two weeks. Our findings show that IL-1 is a key cytokine driving matrix degradation in the degenerate IVD. Furthermore, IL-1Ra delivered directly or by gene therapy inhibits IVD matrix degradation. IL-1Ra could be used therapeutically to inhibit degeneration of the IVD.