白细胞介素1受体拮抗剂对大鼠角膜移植片和房水中炎性细胞因子的影响

Effects of IL-1 receptor antagonist on the level of cytokine in the rat corneal grafts and aqueous humor after corneal transplantation

目的 探讨炎性细胞因子在角膜移植免疫排斥反应中的作用及白细胞介素受体拮抗剂(IL 1ra)的治疗效果。方法 角膜移植大鼠随机分为 5组 ,每组 1 0只 ,采用免疫组化方法和酶链免疫吸附试验 ,检测各组大鼠角膜和角膜移植排斥反应术前、急性排斥反应期及术后 2周植片中白细胞介素 1受体 1 (IL 1RⅠ )和转化生长因子 β1 (TGF β1 )表达情况及房水中白细胞介素 1 β(IL 1 β)含量变化。结果 正常大鼠角膜中未检测出IL 1RⅠ表达 ,TGF β1 主要位于角膜上皮细胞基底层 ;在角膜移植术后的植片上皮细胞层、基质层和内皮细胞层中均可检测到IL 1RⅠ和TGF β1 表达 ,其表达量按阴性对照组、50 μgIL 1ra组、1 0 0 μgIL 1ra组、2 0 0 μgIL 1ra组和地塞米松组依次递减 ,在急性排斥期 ,2 0 0 μgIL 1ra组表达IL 1RⅠ和TGF β1 的程度低于 50 μgIL 1ra组 ,差异有显著意义(P <0 0 1 ) ;阴性对照组IL 1RⅠ和TGF β1 表达量显著高于实验各组 (P <0 0 1 ) ;正常大鼠房水中检测出IL 1 β含量为 (96 0± 1 1 3)ng L ,角膜移植术后房水中IL 1 β含量上升 ,于急性排斥期达到高峰 ,阴性对照组为 (552 2± 68 3)ng L ,明显高于其他各实验组 (P <0 0 1 ) ;在各实验组中 ,排斥前和急性排斥期之间差异无显著意义 (P >0 0

Objective By detecting the expression of IL 1RI and TGF β 1 on the normal rat cornea and graft, and the amount of IL 1β in the aqueous humor of normal rat eye and the eye after keratoplasty, to investigate the relationship between these cytokines and graft rejection and to observe the effects of IL 1 receptor antagonist (IL 1ra) on graft rejection Methods All rats after keratoplasty were divided into five groups Immunohistochemistry method and enzyme linked immunosorbent assay (ELISA) were used to detect the expression of IL 1RⅠ and TGF β 1 on the normal rat cornea and graft and the amount of IL 1β in the aqueous humor of normal rat eye and the eye after keratoplasty at different time points: pre rejection, acute rejection and two weeks after surgery Results IL 1RⅠ could be detected in normal rat cornea TGF β 1 expressed mainly in the epithelium of normal cornea, especially the basal cell layer and the basement membrane After keratoplasty, IL 1RⅠ and TGF β 1 could be detected in the corneal epithelium, stroma and endothelium, and the level of expression decreased in sequence as negative control group, 50 μg IL 1ra group, 100 μg IL 1ra group, 200 μg IL 1ra group and dexamethasone group In the acute rejection period, the expression of IL 1RⅠ and TGF β 1 in the 200 μg IL 1ra group was less than that of the 50 μg IL 1ra group, the difference was significant ( P <0 01) The expression of IL 1RⅠ and TGF β 1 in the negative control group was higher than that of all experi mental groups, the difference was significant ( P <0 01) IL 1β was detected in normal aqueous humor; the mean level was (96 0±11 3) ng/L . In the eye after keratoplasty, the IL 1β level increased significantly compared to normal aqueous humor ( P <0 01) In the acute rejection period, the quantity of IL 1β reached its peak at (552 2±68 3) ng/L in the negative control group, which was the highest quantity of IL 1β in all experimental groups ( P <0 01). The IL 1β level in all experimental groups in the pre rejection period had no difference compared with that in the acute rejection period ( P >0 05), but the level of IL 1β in the pre rejection and rejection periods was significantly different compared with that in the post rejection period ( P <0 01) ConclusionsIL 1RⅠ and TGF β 1 play a active role in the corneal graft immunogenic rejection IL 1β is a key factor in starting corneal graft rejection The keratoplasty graft rejecting reaction can be reduced and mean survival time can be prolonged by IL 1ra, which inhibits the expression of IL 1RⅠ and TGF β 1 and decreases the level of IL 1β in the aqueous humor