IL-1ra对大鼠高危角膜植片IL-1β影响的实验研究

Experimental study of IL-1 receptor antagonist on IL-1β in rat corneal grafts after high-risk penetrating keratoplasty

目的探讨炎性细胞因子白细胞介素-1(interleukin-1,IL-1)β在高危角膜移植免疫排斥反应中的作用及IL-1受体拮抗剂(IL-1 receptor antagonist,IL-1 ra)抑制大鼠高危角膜移植免疫排斥反应的作用机制。方法诱导新生血管后的大鼠行穿透性角膜移植术,受体鼠随机分为3组:生理盐水对照组、10g·L-1环孢霉素A(cyclosporin A,CsA)滴眼液治疗组、50mg·L-1的IL-1ra滴眼液治疗组,每组15只。分别采用原位杂交和免疫组织化学方法检测术后不同时间点各组大鼠角膜移植物IL-1β mRNA及蛋白表达情况。结果正常大鼠角膜IL-1β mRNA在上皮细胞基底层微量表达,IL-1β蛋白无表达;术后不同时间点各移植组角膜植片上皮层、基质层均可检测到IL-1β mRNA及蛋白的阳性表达。术后同一时间点不同移植组相比:IL-1β mRNA及蛋白的表达依次减低顺序为生理盐水对照组、10g.L-1CsA组、50mg·L-1IL-1ra组,差异有显著统计学意义(P<0.01);在急性排斥期,与10g·L-1CsA滴眼液组相比,IL-1ra组降低更明显,两治疗组间比较差异有显著统计学意义(P<0.01)。结论IL-1ra可通过影响IL-1β的表达抑制高危角膜移植免疫排斥反应,减少新生血管生成,减轻免疫性炎性反应,延长角膜植片的存活时间。

Objective To detect the expression of interleukin-1β（IL-1β） and the inhibiting mechanisms of interleukin receptor antagonist （IL-1ra） on the rat cornea graft after high-risk penetrating keratoplasty.Methods Spregue Dawley rats were treated with corneal vascularization,and developed the high risk penetrating keratoplasty.All the models were divided into three groups：control group with normal sodium,treated group with 10 g·L^-1 cyclosporin（CsA） and treated group with 50 mg·L^-1 IL-1ra,15 rats in each group.In situ hybridization and immunohistochemistry method were used to detect the expression of IL-1β mRNA and IL-1β protein at different times in each group.Results IL-β mRNA can be uraced in basal layer of the corneal epithelium in normal rat cosnea,but not IL-1β protein.After high-risk keratoplasty,both the expression of IL-β mRNA and IL-1β protein were detected in the corneal epithelium,stroma and endothelium at different times.Different groups compared at the same time of post-operation：The level of expression of IL-1β mRNA and IL-1β protein decreased in sequence as control group,10 g·L^-1 CsA group and 50 mg·L^-1 IL-1ra group（P〈0.01）;In the acute-rejection period,the expression of IL-1β mRNA and protein in the 50 μg IL-1ra group was less than that of the 10 g·L^-1 CsA group（P〈0.01）.Conclusion IL-1ra can influence the expression of IL-1β,which plays an active role in the cornea graft immunogenic rejection.And it can induce the corneal vascularization,release immunity inflammatory reaction and prolong the survival time of corneal implant.