巨噬细胞炎性蛋白2及其受体在假体无菌性松动周围组织及外周血中的表达及临床意义

The clinical significance of the expressions of chemokine(C-X-C motif) ligand 2 and its receptor in peripheral blood and peri-implant tissues in patients who underwent aseptic implant loosening after total hip replacement

[目的]通过与正常髋关节患者比较,探讨人工髋关节置换术后假体无菌性松动周围组织及外周血中炎性因子巨噬细胞炎性蛋白2（Chemokine（C-X-C motif）ligand 2,CXCL2）及其受体CXC趋化因子受体2（C-X-C chemokine receptor type 2,CXCR2）的表达变化,分析其与无菌性松动间的关系。[方法]以2008年1月～2013年1月22例人工髋关节置换术后假体无菌性松动行人工髋关节翻修术患者作为试验组,20例因其他原因行人工髋关节翻修术患者作为对照组。两组患者年龄及性别比较,差异无统计学意义（P〉0.05）。取两组患者清晨空腹抗凝外周血,以及髋关节翻修术时取假体（或）骨周围组织,采用荧光定量PCR、Western blot以及ELISA法检测CXCL2、CXCR2基因mRNA、蛋白的表达程度及其浓度。免疫组化染色检测CXCL2、CXCR2在组织中表达。[结果]荧光定量PCR检测示,试验组CXCL2基因mRNA在外周血及周围组织中相对表达量（2-ΔΔCt）分别为（18.39±1.11）,（8.07±1.18）,显著高于对照组的（1.00±0.18）（t=119.47,P〈0.01）及（1.00±0.02）（t=45.50,P〈0.01）。CXCR2基因mRNA在外周血及周围组织中相对表达量（2-ΔΔCt）分别为（18.51±1.45）,（8.40±1.02）,显著高于对照组的（1.00±0.09）（t=91.87,P〈0.01）及（1.00±0.02）（t=55.40,P〈0.01）。Western blot检测示,试验组CXCL2蛋白在外周血及周围组织中相对表达比分别为（0.41±0.13）,（0.97±0.24）,显著高于对照组的（0.17±0.07）（t=12.83,P〈0.01）及（0.37±0.09）（t=17.947,P〈0.01）。试验组CXCR2蛋白在外周血及周围组织中相对表达比分别为（0.96±0.30）,（1.15±0.29）,显著高于对照组的（0.64±0.20）（t=6.91,P〈0.01）及（0.52±0.10）（t=15.38,P〈0.01）。ELISA检测外周血CXCL2浓度示,试验组为（3.26±0.59）ng/ml,显著高于对照组的（1.69±0.40）ng/ml（t=17.11,P〈0.01）。免疫组化染色显示松动的假体周围组织中CXCL2、CXCR2表达均显著高于对照组。[结论]CXCL2和CXCR2表达增高可能与人工髋关节置换术后假体无菌性松动有关。

[Objective] To compare the expressions of chemokine（ C- X- C motif） ligand 2 and its receptor in peripheral blood and peri- implant tissues between patients who underwent aseptic implant loosening after total hip replacement and patients who underwent femoral neck fracture,and to analyze the correlation between these expressions and aseptic implant loosening.[Methods] Between January 2008 and January 2013,peripheral blood and tissue samples were collected from 22 patients who underwent aseptic implant loosening after total hip replacement and from 20 patients who underwent revision total hip arthroplasty with other factors in our hospital. The two groups were well matched,without statistically significant differences in age and sex（ P 0. 05）. The mRNA and protein expressions of CXCL2 and CXCR2 were respectively evaluated by using quantitative real-time reverse transcription polymerase chain reaction and Western blot. In addition,the CXCL2 concentration was measured by using enzyme- linked immunosorbent assay（ ELISA）. The peri- implant tissues were evaluated for CXCL2 and CXCR2 expressions by using immunohistochemistry. [Results] The mRNA expression levels（ 2- ΔΔCt） of CXCL2 in peripheral blood and peri- implant tissues in the patients who underwent aseptic implant loosening after total hip replacement were（ 18. 39 ± 1. 11） and8. 07 1. 18 ng / m L,respectively,which were significantly higher than those in the control group,which were（ 1. 00 ± 0. 18） ng /m L（ t = 119. 47,P 0. 01） and（ 1. 00 ± 0. 02） ng / m L（ t =45. 50, P 0. 01）, respectively. The mRNA expression levels（ 2- ΔΔCt） of CXCR2 in the peripheral blood and peri- implant tissues of the patients who underwent aseptic implant loosening after total hip replacement were（ 18. 51 ± 1. 45） and（ 8. 40 ± 1. 02） ng / m L,respectively,which were significantly higher than those in the control group,which were（ 1. 00 ± 0. 09） ng / m L（ t = 91. 87,P 0. 01） and（ 1. 00 ± 0. 02） ng / m L（ t = 55. 40,P 0. 01）,respectively. The relative ratios of CXCL2 protein expression in peripheral blood and peri- implant tissues in the experimental group were（ 0. 41 ± 0. 13） and（ 0. 97 ± 0. 24）,respectively,which significantly higher than those in the control group,which were（ 0. 17 ± 0. 07）（ t = 12. 83,P 0. 01） and（ 0. 37 ± 0. 09）（ t = 17. 947,P 0. 01）,respectively. The relative ratios of CXCR2 protein expression in peripheral blood and peri- implant tissues in the experimental group were（ 0. 96± 0. 30） and（ 1. 15 ± 0. 29）,respectively,which were significantly higher than those in the control group,which were（ 0. 64± 0. 20）（ t = 6. 91,P 0. 01） and（ 0. 52 ± 0. 10）（ t = 15. 38,P 0. 01）,respectively. The results from the ELISA showed that the CXCL2 concentration in the experimental group was significantly higher than that in the control group（ 3. 26 ±0. 59 ng / m L vs（ 1. 69 ± 0. 40） ng / m L; t = 17. 11,P 0. 01）. The positivity rates of the CXCL2 and CXCR2 expressions in the peri- implant tissues in the experimental group were significantly higher than those in the control group by immunohistochemistry. [Conclusion] The higher CXCL2 and CXCR2 expression levels in peripheral blood in the patients who underwent total hip replacement are probably correlated with aseptic implant loosening,which can thus be considered as a prognostic factor.