Downregulation of Hypoxia-Inducible Factor-1α by RNA Interference Alleviates the Development of Collagen-Induced Arthritis in Rats
Downregulation of Hypoxia-Inducible Factor-1α by RNA Interference Alleviates the Development of Collagen-Induced Arthritis in Rats
复制标题
RNA 干扰下调缺氧诱导因子 1 α 可减轻大鼠胶原诱导的关节炎的发展
DOI：
10.1016/j.omtn.2020.01.014
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发表时间：
2020-03-06
影响因子：
8.8
通讯作者：
Zhang, Peng
中科院分区：
文献类型：
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作者：
Hu, Yiping;Zhang, Tiantian;Zhang, Peng
类风湿性关节炎（RA）是最常见的自身免疫性关节炎。缺氧诱导因子-1 α（Hypoxia-inducible factor-1 alpha，HIF-1 alpha）作为一种缺氧应答的转录因子，可能成为治疗类风湿关节炎的潜在靶点。在这项研究中，我们评估了HIF通路阻断是否减轻了胶原诱导性关节炎（CIA）大鼠模型中RA的表现。我们构建了靶向HIF-1 α的短发夹状RNA（shRNA）慢病毒表达载体（pLVX-shRNA-HIF-1 α）并实现了HIF-1 α RNA干扰。采用定量RT-PCR、免疫荧光染色和western blot分别检测HIF-1 α、血管内皮生长因子（VEGF）、phsopho（p）-p65和p-IKB α mRNA和蛋白的表达。在CIA诱导后的不同时间点，使用微型计算机断层扫描来研究关节形态。酶联免疫吸附试验（ELISA）检测炎症细胞因子的表达。体外分析显示，pLVX-shRNA-HIF-1 α有效抑制HIF-1 α和VEGF的表达，并导致p-65和p-IKB α的活化，以及细胞培养物中促炎细胞因子表达的降低。在大鼠中抑制HIF-1 α减少了全身炎症状况的迹象，同时减少了RA的病理变化。此外，下调HIF-1 α表达显著减少滑膜炎和血管生成。总之，我们已经表明，药物抑制HIF-1可能会改善RA的临床表现。
Rheumatoid arthritis (RA) is the most common type of autoimmune arthritis. Hypoxia-inducible factor-1 alpha (HIF-1 alpha) as a transcription factor in response to hypoxia suggests that it could be a potential therapeutic target for the treatment of RA. In this study, we assessed whether the HIF pathway blockade attenuates the manifestations of RA in the collagen-induced arthritis (CIA) rat model. We constructed a short hairpin RNA (shRNA) lentiviral expression vector targeting HIF-1 alpha (pLVX-shRNA-HIF-1 alpha) and to achieve HIF-1 alpha RNA interference. Quantitative RT-PCR, immunofluorescence staining, and western blot were used to detect the expressions of HIF-1 alpha, vascular endothelial growth factor (VEGF), phsopho (p)-p65, and p-IKB alpha mRNA and protein, respectively. Micro-computed tomography was used to investigate joint morphology at different time points after CIA induction. Moreover, enzyme-linked immunosorbent assay (ELISA) was used to monitor the expression of inflammatory cytokines. In vitro analyses revealed that pLVX-shRNA-HIF-1 alpha effectively inhibited the expression of HIF-1 alpha and VEGF and led to the activation of p-65 and p-IKB alpha, as well as decreased proinflammatory cytokine expression in cell culture. Inhibition of HIF-1 alpha in rats decreased signs of a systemic inflammatory condition, together with decreased pathological changes of RA. Moreover, downregulation of HIF-1 alpha expression markedly reduced the synovitis and angiogenesis. In conclusion, we have shown that pharmacological inhibition of HIF-1 may improve the clinical manifestations of RA.