核受体RARr选择性激动剂CD437抑制IL-1β诱导的角膜基溶解实验研究

The inhibition effect of Nuclear RAR-r selective agonist CD437 on corneal matrix degradation induced by IL-1β

目的角膜溃疡的形成与MMPs诱导的角膜组织的过度溶解有关,本实验拟研究RARr选择性激动剂CD437抑制IL-1β诱导的角膜基质角膜基质胶原降解的机制。方法兔角膜基质细胞分离培养后,与无血清培养液,Ⅰ型胶原,5×DMEM于冰上混合,然后置于培养箱中凝固。然后将不同浓度的CD437与纤溶酶原混合液被覆于胶原表面,收集培养上清液,检测上清液中的羟脯氨酸含量,免疫印迹法检测MMP1,3表达量,明胶酶谱法检测MMP2,9表达量。结果 CD437可抑制IL-1β诱导的角膜基质胶原降解,呈剂量及时间依赖关系,CD437可抑制IL-1β诱导的前体及活性MMP1,2,3,9表达量。结论 CD437作为选择性RTRAr激动剂,可抑制IL-1β诱导的角膜基质胶原降解,具有临床治疗角膜溃疡潜力。

Objective Corneal ulcer was based on cornea tissues massive resolution induced by MMPs.This research focus on the inhibition effect of RAR r agonist CD437in corneal collagen degradation induced by IL-β.Methods Rabbit corneal fibroblasts were cultured in three-dimensional collagen gels with Serum free medium,type I collagen,5×DMEM on ice until collagen solidification in incubator.Different concentration of CD437and plasminogen were put on the surface of collagen,Collection of the culture supernatant,The HYP amount was messured.The expression and activity of MMP 1,2,3,9were evaluated by immunoblot analysis and gelatin zymography.Results CD437inhibit corneal collagen degradation induced by IL-1βin a concentration and time-dependent manner.MMP1,2,3,9expression and activation exposed to IL-1βwere also inhibited by CD437.Conclusion As selective RAR r agonist,CD437can inhibit IL-1β– stimulated coneal collagen degradaation thereby be potent in the therapy of corneal ulcer.