白细胞介素-1受体拮抗蛋白对大鼠创伤性骨关节炎的治疗作用及机制研究

Effect and mechanism of interleukin-1 receptor antagonist protein on post-traumatic osteoarthritis in rats

目的研究白细胞介素-1受体拮抗蛋白(IL1RA)对大鼠创伤性骨关节炎(PTOA)的治疗作用及机制。方法将50只SD大鼠分为假手术组,模型组,IL1RA低、中、高剂量组(10、20、40mg/mL),除假手术组外,其余组均复制PTOA模型。ELISA检测白细胞介素(IL)-1β、IL-8、肿瘤坏死因子-α(TNF-α)水平,Realtime PCR法检测Ⅱ型胶原蛋白(COL-Ⅱ)、蛋白聚糖(Aggrecan)、基质金属蛋白酶-1(MMP-1)、MMP-13、聚蛋白多糖酶-4(ADAMTs-4)及ADAMTs-5 mRNA的表达,Western blot检测MMP-1、MMP-13、ADAMTs-4、ADAMTs-5、基质金属蛋白酶抑制剂-1(TIMP-1)及TIMP-3蛋白表达。结果与假手术组比较,模型组IL-1β、IL-8及TNF-α水平明显提高(P<0.05),COL-Ⅱ、Aggrecan mRNA表达明显下调(P<0.05),MMP-1、MMP-13、ADAMTs-4、ADAMTs-5mRNA及蛋白表达明显上调(P<0.01),TIMP-1及TIMP-3蛋白表达明显下调(P<0.05)。与模型组比较,IL1RA低、中、高剂量组IL-1β、IL-8及TNF-α水平明显降低(P<0.05),COL-Ⅱ、Aggrecan mRNA表达明显上调(P<0.05),MMP-1、MMP-13、ADAMTs-4、ADAMTs-5mRNA表达明显下调(P<0.05),ADAMTs-4、ADAMTs-5蛋白表达明显下调(P<0.05),TIMP-1及TIMP-3蛋白表达明显上调(P<0.05);IL1RA中、高剂量组中MMP-1及MMP-13蛋白表达明显下调(P<0.05)。结论IL1RA对PTOA大鼠具有治疗作用,与抑制细胞外基质降解,促进细胞外基质合成有关。

Objective To investigate effect and mechanism of interleukin-1 receptor antagonist protein(IL1 RA)on post-traumatic osteoarthritis(PTOA)in rats.Methods Fifty SD rats were randomly divided into the sham operation group,model control group,IL1 RA low,medium and high doses groups(10,20,40 mg/mL).Except for the sham operation group,the PTOA model was copied in the other groups.The level of IL-1β,IL-8 and tumor necrosis factor-α(TNF-α)was detected by using the enzyme-linked immunosorbent assay(ELISA).The expressions of type Ⅱ collagen(COL-Ⅱ),Aggrecan,matrix metalloproteinase-1(MMP-1),MMP-13,a disintegrin-like and metalloproteinase with thrombospondin motifs-4(ADAMTs-4)and ADAMTs-5 mRNA were detected by the realtime PCR method.The expression of MMP-1,MMP-13,ADAMTs-4,ADAMTs-5,tissue inhibitor of metalloproteinases-1(TIMP-1)and TIMP-3 protein was detected by Western blot.Results Compared with the sham operation group,the levels of IL-1β,IL-8 and TNF-αin the model group were significantly increased(P<0.05),the expressions of COL-Ⅱ and Aggrecan mRNA were significantly down-regulated(P<0.05),the expressions of MMP-1,MMP-13,ADAMTs-4 and ADAMTs-5 protein and mRNA were significantly up-regulated(P<0.05),the expressions of TIMP-1 and TIMP-3 protein were significantly down-regulated(P<0.05).Compared with the model group,the levels of IL-1β,IL-8 and TNF-αin the ILIRA low,medium and high doses groups were significantly decreased(P<0.05),the expressions of COL-Ⅱ and Aggrecan mRNA were significantly up-regulated(P<0.05),while the expressions of MMP-1,MMP-13,ADAMTs-4 and ADAMTs-5 mRNA were significantly down-regulated(P<0.05),the expressions of ADAMTs-4 and ADAMTs-5 protein were significantly down-regulated(P<0.05),the expressions of TIMP-1 and TIMP-3 protein were significantly up-regulated(P<0.05);the expressions of MMP-1,MMP-13 protein in the IL1 RA medium and high doses groups were significantly down-regulated(P<0.05).ConclusionIL1 RA has the curative effect on PTOA rats,which may be related with inhibiting extracellular matrix decomposition and promoting the synthesis of extracellular matrix.