白介素-1受体拮抗剂对双后肢大鼠椎间盘细胞凋亡的影响

The effects of interleukin-1 receptor antagonist on the apoptosis of the intervertebral disc cell in bipedal rat

目的:探讨白介素-1受体拮抗剂(IL-1Ra)对双后肢大鼠椎间盘细胞凋亡的影响。方法:建立双后肢大鼠模型54只,随机分为试验组、空白对照组和阴性对照组,每组18只,试验组从造模当天即予腹腔注射IL-1Ra(50ng/kg体重),隔天1次重复注射至处死,空白对照组不予任何处理,阴性对照组隔天1次予腹腔注射等量生理盐水。于造模后1、3、6个月每组分别处死6只大鼠,完整取出L4/5椎间盘后进行固定、切片,使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法(TUNEL法)检测各组大鼠椎间盘细胞的凋亡情况,并使用透射电镜观察凋亡细胞的超微结构。取L5/6椎间盘使用流式细胞仪检测各组大鼠椎间盘细胞的凋亡情况。结果:术后1个月试验组、空白对照组及阴性对照组椎间盘细胞的凋亡无显著性差异(P>0.05);术后3个月试验组与空白对照组及阴性对照组相比细胞凋亡明显减少,有显著性差异(P<0.05);术后6个月试验组与空白对照组及阴性对照组相比细胞凋亡率有非常显著性差异(P<0.01)。结论:IL-1Ra对双后肢大鼠椎间盘细胞凋亡有明显的抑制作用。

Objective：To explore the effects of the interleukin-I receptor antagonist protein（IL-1Ra） on the apoptosis of the bipedal rats＇ intervertebral disc cell.Method：The degenerative model of bipedal rats was established.The 54 rats were randomly divided into three groups：experimental group,blank-control group and negative-control group with each group 18 rats.The experimental group rats were intraperitoneal injected （50ng/kg） with IL-1Ra from the day of operation and repeated on alternate days until sacrefised,and in the negative-control group 0.9% NS of the same dosage of IL-1Ra were injected.Nothing was injected into the rats of the blank-control group.Six rats were sacreflsed from each group at the time of one,three,six month post-operatively respectively.The tissue of IA/5 discs en bloc were fixed and sliced.The apoptosis cells in the intervertebral disc were assayed using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL） and the ultranstructure were examined by transmission electron microscopy;and the tissue of the L5/6 discs were assayed using flowcytometry.Result：There were no significant difference in terms of the apoptosis cells in intervertebral disc among these 3 groups at 1 month posteratively（P〉0.05）.The apoptosis cells in the experimental group decreased remarkably than those in blank-control group and negative-control group at three month postoperatively（P〈0.05）,which was the same as those at six month postoperatively（P〈0.01）.Condusion：IL-IRa protein can inhibit the cell apoptosis of the intervertebral disc in bipedal rat.