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RNAi下调MMP-14表达对类风湿关节炎滑膜细胞炎症因子分泌的影响 被引量：5

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摘要 目的探究能否通过RNA干扰(RNAi)沉默膜型基质蛋白酶-14(MMP-14)从而抑制类风湿关节炎患者的成纤维滑膜细胞(RA-FLS)炎症因子的分泌。方法体外化学合成MMP-14序列特异性双链siRNA,与脂质体混合后转染RA-FLS,实验分为3组,MMP-14-siRNA组转染MMP-14特异性siRNA的RA-FLS,No target组转染MMP-14非特异性siRNA的RA-FLS,Control组为未经处理的RA-FLS。用实时荧光定量逆转录聚合酶链反应(QRT-PCR)检测相关基因转录水平,蛋白质免疫印迹法(western blot)检测MMP-14蛋白水平,Human IL-1βimmunoassay检测试剂盒检测细胞炎症因子IL-1β分泌。结果 QRT-PCR结果显示,MMP-14-siRNA组细胞中MMP-14基因表达0.290±0.026,明显比No target组和Control组低(P<0.001),而No target组细胞的MMP-14基因表达0.984±0.016与Control组比较差异无统计学意义(P>0.05)。RNAi抑制MMP-14基因转录后,MMP-14-siRNA组细胞MMP-14蛋白表达水平明显低于Control组和No target组。QRT-PCR结果显示MMP-14-siRNA组的炎症因子TNF、IL-1β基因转录水平明显降低,分别为Control组的73%和52%,而COX-2的表达差异无统计学意义(P>0.05)。Human IL-1βimmunoassay检测试剂盒检测结果进一步验证了MMP-14-siRNA组细胞向胞外释放的IL-1β显著减少,约为Control组的50%,而No target组与Control组差异无统计学意义(P>0.05)。结论使用RNA干扰技术可有效抑制RA-FLS炎症因子分泌,提示RNAi技术沉默MMP-14有可能缓解RA滑膜炎症。
作者 刘振杰 张丹丹 李涛 徐宁 李曼 黄丽英
机构地区 广东省中医院芳村医院检验科
出处 《广东医学》 CAS 北大核心 2015年第15期2342-2345,共4页 Guangdong Medical Journal
关键词 RNA干扰 MMP-14 类风湿关节炎 成纤维滑膜细胞 炎症因子
分类号 R736.1 [医药卫生—肿瘤]
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参考文献18

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二级参考文献24

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共引文献30

同被引文献43

引证文献5

二级引证文献45

广东医学
2015年 第15期

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