白介素-1受体拮抗蛋白的应用对早期创伤性关节炎软骨细胞代谢的影响  

Early Intervention with Interleukin-1 Receptor Antagonist Protein Modulates Cell Metabolism in Cartilage after Impact Injury

作　　者：谷崎[1] 严春梅[2] 夏挺 GU Qi YAN Chunmei XIA Ting(Xi＇an Technological University, Xi＇an 710032, China School of Physical Education, Kunming University, Kunming 654300, China Department of Critical Care Medicine, Weinan First Hospital, Weinan 714000,China)

机构地区：[1]西安工业大学高水平运动管理中心,西安710032 [2]昆明学院体育学院,昆明654300 [3]渭南市第一医院重症医学科,陕西渭南714000

出　　处：《西安体育学院学报》2017年第4期482-487,495,共7页Journal of Xi'an Physical Education University

摘　　要：目的探讨应用白介素-1受体拮抗蛋白(IL1RA)对早期创伤性关节炎软骨细胞代谢的影响,主要观察应用后相关细胞炎性因子及蛋白酶基因的表达变化。方法 44只健康成年新西兰大白兔,随机分为对照(假手术)组(CON组,n=4)、安慰剂组(PBS组,n=20)、IL1RA治疗组(IL1RA组,n=20)。其中PBS组及IL1RA组均接受关节内重物击打,造成创伤性关节炎模型。造模后1 h,CON组不接受任何治疗,PBS组及IL1RA组实验兔膝关节则分别给予0.5 m L无菌PBS和0.5 m L IL1RA-PBS关节腔注射。术后3 h及8 h采集关节液,检测IL-1β含量。术后8 h处死动物取软骨组织进行软骨细胞培养后,测定细胞活力及凋亡率,RT-PCR检测软骨组织中ADAMTs-4、ADAMTs-5、MMP-3、IL-1β及TNF-α的m RNA表达。结果术后3 h及8 h,PBS组关节液中IL-1β的含量均明显高于CON组(P<0.01)。然而,经IL1RA介入治疗后,IL-1β在术后各测试点的含量均明显低于PBS组(P<0.05)。与CON组相比,术后8h培养的PBS组软骨细胞活力明显降低(P<0.05),而IL1RA组软骨细胞活力与CON组相比无显著性差异,但明显高于PBS组。另一方面,与CON组及IL1RA组相比,PBS组的细胞凋亡率明显增高(P<0.05)。而IL1RA组与CON组相比,细胞凋亡率无明显差别(P>0.05)。RT-RCR结果显示,与CON组比较,PBS组所有目的基因m RNA的表达均明显增加(P<0.01)。然而,IL1RA组所有目的基因的表达与CON组相比无统计学差异,但明显低于PBS组(P<0.01)。结论实验结果表明创伤后关节炎早期应用IL1RA介入治疗可有效降低关节液细胞炎性因子的浓度,抑制细胞炎性因子对软骨细胞生长增殖的负性影响,降低细胞凋亡,且对于促进软骨细胞基质降解的细胞炎性因子及蛋白酶的表达显著下调,对PTOA病程发展具有抑制作用。Objective The purpose of this study was to determine the efficacy of Intcrleukin-1 Receptor Antagonist Protein （ILl RA） treatment as an early intervention strategy by examining the changes in cell metabolism in cartilage in a rabbit knee joint impact model. Methods Custom impact device was used to create injury to intact rabbit knee joint. Injury was caused by dropping a 0. 43 kg weight one time from 20cm directly above the knee in flexion. 1 hour after impact PBS solution or 10 mg/mL IL1RA solution was intra-articularly injected. At 3h and 8h post-injury, the joint fluid were collected to detect the content of IL-1β. At 8 h post-injury, cartilage samples were harvested for cell viability, cell apoptosis and genetic expression analysis. Genetic expression of ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-a were analyzed by RT-PCR. Results At 3 h and 8 h post-injury, The IL-1β eontent in PBS group were significantly higher than those in CON group （P 〈 0. 01 ）. After IL1RA intervention, the IL-1β contents of each testing time-point after surgery were significantly lower than those in the PBS group （P 〈 0.05 ）. Compared with CON group, cell activity in PBS group were significantly decreased （P 〈 0. 05 ）, and compared with CON group, cartilage cell vitality of ILl RA group had no significant decrease, but was significantly higher than the PBS group. On the other hand, compared with CON and ILIRA group, cell apoptosis rate of PBS group was the highest （P 〈 0. 05 ）. At 8 h after ILl RA treatment, expressions of ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-α in cartilage were significantly down-regulated from PBS group （ all P 〈 0.01 ）. Conclusion This study demonstrated that IL1RA treatment administered during acute phase of cartilage impact injury decreased the IL-1β content of the joint fluid, inhibited the negative effects of inflammatory factors on the cell growth and vitality, decreased expression levels of ADAMTS-4, ADAMTS-5, MMP-3, IL-1β, and TNF-α in cartilage. Clinica

关 键 词：白介素-1 受体拮抗蛋白早期介入 创伤性关节炎 软骨细胞代谢 

分 类 号：G804.5[文化科学—运动人体科学]