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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: To investigate the effect of interleukin-1 receptor antagonist (IL1RA) on the metabolism of chondrocytes in patients with early traumatic arthritis, and to observe the changes of the related inflammatory cytokines and protease genes. Methods 44 healthy adult New Zealand white rabbits were randomly divided into control group (CON group, n = 4), placebo group (n = 20 in PBS group) and IL1RA group (n = 20) . Among them, PBS group and IL1RA group received intra-articular weight hitting, resulting in a traumatic arthritis model. One hour after modeling, the CON group did not receive any treatment. The rabbits in the PBS group and the IL1RA group were injected with 0.5 mL sterile PBS and 0.5 mL IL1RA-PBS intra-articularly. The synovial fluid was collected 3 h and 8 h after operation, and the content of IL-1β was detected. After 8 h, the cartilage tissues were sacrificed and the chondrocytes were cultured. The viability and apoptosis rate of chondrocytes were measured. The expression of ADAMTs-4, ADAMTs-5, MMP-3, IL-1 and TNF- m RNA expression. Results The levels of IL-1β in synovial fluid of PBS group were significantly higher than those of CON group at 3 h and 8 h after operation (P <0.01). However, the levels of IL-1β at each test point after IL1RA intervention were significantly lower than those in PBS group (P <0.05). Compared with CON group, the viability of chondrocytes in PBS group was significantly decreased 8 h after operation (P <0.05), while the activity of chondrocytes in IL1RA group was not significantly different from that in CON group, but significantly higher than that in PBS group. On the other hand, compared with the CON group and IL1RA group, the apoptosis rate of PBS group was significantly higher (P <0.05). There was no significant difference in apoptosis rate between IL1RA group and CON group (P> 0.05). The results of RT-PCR showed that compared with the CON group, all the m RNA expression of all the target genes in the PBS group increased significantly (P <0.01). However, there was no significant difference in the expression of all the target genes in IL1RA group compared with CON group, but significantly lower than that in PBS group (P <0.01). Conclusion The experimental results show that the early application of IL1RA interventional therapy in post-traumatic arthritis can effectively reduce the concentration of inflammatory cytokines in the synovial fluid and inhibit the negative effects of cytokines on the proliferation and proliferation of chondrocytes and reduce the apoptosis of chondrocytes. Matrix degradation of inflammatory cytokines and protease expression was significantly down-regulated PTOA course of development has an inhibitory effect.