Analysis of distribution, phenotype and clinical correlation of CD163 positive macrophages in osteoarthritis
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摘要:
目的 分析骨关节炎(osteoarthritis，OA)患者CD163+巨噬细胞在滑膜及外周血的分布特征，探讨CD163+巨噬细胞及胞内肿瘤坏死因子(tumour necrosis factor，TNF)、白细胞介素-10(interleukin-10，IL-10)表达水平与OA病程、炎症水平、体重指数(body mass index，BMI)的相关性。 方法 流式细胞术检测OA患者滑膜及外周血CD163+巨噬细胞及胞内TNF-α+、IL-10+表达水平；免疫荧光观察OA患者滑膜中CD163+巨噬细胞。 结果 (1) OA患者和骨折患者外周血和滑膜中CD163+巨噬细胞表达差异均有统计学意义(均有P < 0.05)。(2)OA病程 < 10年组患者滑膜及外周血中CD163+巨噬细胞内TNF-α+表达比例明显高于OA病程≥ 10年组及骨折组(均有P < 0.05)；OA病程 < 10年组患者滑膜及外周血中IL-10+表达比例明显低于OA病程≥ 10年组及骨折组(均有P < 0.05)(3)OA病程 < 10年组患者C-反应蛋白(C-reactive protein，CRP)与BMI呈正相关(r=0.680，P < 0.001)，OA病程≥ 10年组患者CRP与BMI不相关(r=0.084，P=0.187)。(4)免疫荧光共聚焦显微镜显示OA患者滑膜组织血管周围有大量CD163+巨噬细胞浸润。 结论 病程不同的OA患者CD163+巨噬细胞在滑膜及外周血中的分布存在差异，并呈现差异化细胞亚型。 Abstract:Objective The aim of this study was to analyze the distribution characteristics of CD163+ macrophages in peripheral blood and synovium of patients with osteoarthritis (OA), and investigate the correlation of CD163+ macrophages, intracellular tumour necrosis factor(TNF-α), interleukin-10 (IL-10) expression with the disease duration, inflammation level and body mass index (BMI). Methods The percentages of CD163+ macrophages in synovium and peripheral blood and their intracellular levels of TNF-α and IL-10 were detected by flow cytometry. Meanwhile, immunofluorescence was uesed to observe the distribution of CD163+ macrophages in synovial tissue of patients with OA or fracture. Results (1) There were significant differences in the percentage of CD163+ macrophages in synovial and peripheral blood between OA and fracture patients(all P < 0.05). (2) The expression of TNF-α in CD163+ macrophages of synovium and blood of OA patients with disease duration below 10 years was significantly higher than that of patients with OA disease duration above 10 years (all P < 0.05). The expression of IL-10 was significantly lower than that of patients with OA disease duration above 10 years (all P < 0.05). (3) C-reactive protein (CRP) was positively correlated with BMI in patients with OA disease duration below 10 years (r=0.680, P < 0.001), while showed no correlation with BMI in patients with OA duration above 10 years(r=0.084, P=0.187). (4) Immunofluorescence confocal microscopy showed a large number of CD163+ macrophages invaded around the synovial vessel of patients with OA. Conclusion The distribution of CD163+ macrophages in synovium and peripheral blood of OA patients with different course of disease was different, and showed differentiated cell subtypes. -
Key words:
- Osteoarthritis /
- Macrophages /
- Inflammation /
- IL-10 /
- TNF-α
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图 1 关节滑膜中CD163+巨噬细胞表达
注：A-C：病程＜10年组，D-F：病程≥10年组，G-I：骨折组。DAPI染色(蓝)、CD163染色(绿)，图像重叠处理：Merge(黄)。
Figure 1. The expression of CD163+ macrophages in synovium
表 1 OA及骨折患者外周血和滑膜中各个指标的差异比较
Table 1. Comparison of differences in peripheral blood and synovial membrane between OA and fracture patients
指标 外周血 滑膜中 t值 P值 CD163+ 27.71±14.31 7.78±6.78 12.734 < 0.001 IL-10+ 3.70±3.31 3.59±3.08 0.318 0.752 TNF-α+ 32.97±11.28 30.03±7.94 2.563 0.014 
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表 2 OA及骨折患者外周血和滑膜中CD163+巨噬细胞及胞内TNF-α+和IL-10+表达
Table 2. Expression of CD163+ macrophages and intracellular TNF-α+ and IL-10+ in peripheral blood and synovium of patients with osteoarthritis and fracture
标本类型 指标 ＜10年组 ≥10年组 骨折组 F值 P值 滑膜 CD163+巨噬细胞 37.34±12.54 21.08±3.63 5.91±0.19 29.985 < 0.001 TNF-α+ 35.54±6.72 23.04±2.71 26.8±1.07 9.397 0.005 IL-10+ 2.21±1.89 3.71±2.11 9.4±3.74 29.936 < 0.001 外周血 CD163+巨噬细胞 12.97±5.16 2.26±1.08 0.51±0.04 22.311 < 0.001 TNF-α+ 41.61±7.62 24.76±3.68 17.5±0.68 57.195 < 0.001 IL-10+ 2.23±2.56 4.82±3.32 7.52±1.29 9.259 0.008
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表 3 CD163+巨噬细胞及胞内TNF-α+与IL-10+水平与炎症指标和BMI相关性
Table 3. Correlation between CD163+ macrophages, intracellular TNF-α+ and IL-10+ levels and inflammatory markers and BMI
指标 < 10年 ≥10年 ESR CRP ESR CRP 外周血 CD163+巨噬细胞 -0.122 -0.151 -0.185 -0.109 IL-10+ -0.203 -0.112 0.065 -0.053 TNF-α+ 0.012 -0.253 -0.137 -0.135 滑膜 CD163+巨噬细胞 0.281 0.023 0.325 0.267 IL-10+ 0.140 -0.100 0.014 -0.105 TNF-α+ 0.063 -0.123 -0.341 -0.367 BMI 0.109 0.680a 0.241 0.084 注：a表示P＜0.05。
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