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| IL-6R抗体调节PMMA骨水泥介导的滑膜成纤维细胞MMPs和血管化因子mRNA表达 |
| Regulation of IL-6R antibody on PMMA bone cement mediated mRNA expression of MMPs and vascular factors by synovial fibroblasts |
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| DOI： |
| 中文关键词: IL-6R抗体 聚甲基丙烯酸甲酯 骨水泥 滑膜成纤维细胞 基质金属蛋白酶-l 基质金属蛋白酶-3 血管内皮生长因子 血小板衍生生长因子 |
| 英文关键词:IL-6R antibody PMMA Bone cement Synovial fibroblasts MMP-1 MMP-3 VEGF PDGF |
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| 中文摘要: |
| 目的 探讨IL-6R抗体对PMMA骨水泥介导的滑膜成纤维细胞MMP-l和MMP-3、VEGF和PDGF mRNA表达的调节。方法 从全髋关节置换术中获取滑膜组织消化并传代培养。倒置相差显微镜对滑膜细胞进行形态学观察，免疫细胞化学(SABC法)染色对滑膜成纤维细胞进行鉴定。根据加入不同培养物，实验分为3组：PMMA组：75μg/mL PMMA骨水泥颗粒；IL-6R抗体组：10 ng/mL IL-6R抗体+75μg／mL PMMA骨水泥颗粒；空白对照组。采用细胞计数试剂盒-8(Cell Counting Kit-8，CCK．8)检测IL-6R抗体、PMMA对滑膜成纤维细胞增殖活力影响；实时荧光定量PCR(real time fluorescence quantitative polymerase chain reaction．FQ-PCR)检测MMP-1、MMP-3、VEGF和PDGF mRNA表达。结果 原代滑膜细胞贴壁后，初期为短梭形。连续3次传代后，95％以上细胞为长梭形成纤维细胞样细胞。SABC法染色检测结果显示：上述成纤维样细胞抗CD68抗体阴性，抗vimentin抗体呈棕黄色，为阳性反应。CCK一8检测显示：与PMMA组和空白对照组相比，IL-6R抗体组吸光度(A)值明显降低(P<0.01)；而空白对照组与PMMA组间吸光度(A)值无统计学差异(P>0.05)。FQ-PCR检测发现：与PMMA组和空白对照组相比，IL-6R抗体组中MMP-l和MMP-3、VEGF和PDGF mRNA的表达明显受到抑制(P<0.01)；PMMA组中MMP-l和MMP-3、VEGF和PDGF表达较空白对照组升高(P<0.05)。结论 IL-6R抗体能显著抑制滑膜成纤维细胞MMP-1和MMP-3、VEGF和PDGFmRNA的表达，为生物制剂预防和治疗假体无菌性松动提供分子生物学的理论依据。 |
| 英文摘要: |
| Abstract：Objective To investigate the PMMA bone cement-mediated expression of MMP-1，MMP-3，VEGF，and PDGF mRNA in synovial fibrublasts regulated by IL-6R antibody．Methods The synovial tissue obtained from total hip arthroplasty was digested and subcuhured．Phase-contract microscope was employed to observe the synovial cells and immunocytochemistry(SABC method)staining was used to identify synovial fibroblasts．The experiment was divided inlo 3 groups according to the different supplement in cultures：PMMA group(75μg/ml PMMA bone cement particles)；IL-6R antibody group(10 ng/ml IL-6R antibody and 75μg/ml PMMA bone cement particles)；and the control group．Cell proliferation activity regulated by IL-6R antibody or PMMA was measured by using a cell counting kit-8(CCK-8)．FQ-PCR was used to detect the mRNA expressions of MMP-1，MMP-3，VEGF，and PDGF．Results The primary synovial ceils showed a short spindle shape after adhesion．After 3 consecutive passages，more than 95％ of the cells became fibroblast-like cells with a long spindle shape．SABC staining showed that the anti-CD68 antibody was negative and anti-vimentin antibody was positive in fibroblast-like cells．CCK-8 test demonstrated that the absorbance value(A)was significantly lower in IL-6R antibody group than that in PMMA group and control group(P<0.01)．The A value was not statistically different between the control group and PMMA group(P>0.05)．FQ-PCR measurement showed that mRNA expressions of MMP-1，MMP-3，VEGF，and PDGF in IL-6R antibody group were significantly inhibited than those in PMMA group and control group(P<0.01)．The mRNA expressions of MMP-1，MMP-3，VEGF，and PDGF in PMMA group were higher than those in control group(P<0.05)．Conclusion IL-6R antibody could significantly inhibit the mRNA expressions of MMP-1，MMP-3，VEGF，and PDGF in synovial fibroblasts．This may provide theoretical basis of molecular biology for the prevention and treatment of aseptic loosening of the prosthesis． |
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