The effect of hyaluronic acid on nucleus pulposus extracellular matrix production through hypoxia-inducible factor-1α transcriptional activation of CD44 under hypoxia
The effect of hyaluronic acid on nucleus pulposus extracellular matrix production through hypoxia-inducible factor-1α transcriptional activation of CD44 under hypoxia
复制标题
透明质酸通过缺氧诱导因子-1α转录激活CD44对髓核细胞外基质产生的影响
DOI：
10.22203/ecm.v041a10
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发表时间：
2021
影响因子：
3.1
通讯作者：
S Liu
中科院分区：
文献类型：
--
作者：
F Zhang;X Liu;B Li;Z Li;S Grad;D Chen;M Gao;S Liu
椎间盘退变（IDD）是导致腰痛的主要原因。透明质酸（HA）植入是治疗IDD的潜在策略，但其在缺氧条件下的药理作用和机制尚不清楚。在这项研究中，细胞外基质的基因和蛋白的表达增强髓核细胞（NPC）在缺氧条件下，HA的存在下，所示的实时逆转录-聚合酶链反应，免疫荧光染色，二甲基亚甲基蓝测定。此外，在HA和缺氧的存在下，与单独的任何一个相比，CD 44的表达增加。缺氧诱导因子-1 α（hypoxia inducible factor-1α，HIF-1α）是缺氧条件下的关键转录因子，利用生物信息学数据库发现HIF-1 α在CD 44启动子上有4个预测的结合位点。用氯化钴或二甲基草酰甘氨酸处理后，CD 44表达显著增加。HIF-1α在NPC中的过表达显著上调了CD 44的表达。通过启动子截短实验和染色质免疫沉淀分析确定了HIF-1α在CD 44启动子区的结合位点。综上所述，这些结果表明，缺氧条件下积极增强的能力，NPCs的基质合成在HA的存在下，这与增加的CD 44表达的HIF-1α转录激活。
Intervertebral disc degeneration (IDD) is the leading cause of low-back pain. Implantation of hyaluronic acid (HA) is potentially a therapeutic strategy for IDD, but its pharmacological effects and mechanism under hypoxic conditions remain unclear. In this study, the expression of extracellular matrix genes and proteins were enhanced in nucleus pulposus cells (NPCs) in the presence of HA under hypoxic condition, as shown by real-time reverse transcription-polymerase chain reaction, immunofluorescence staining, and dimethylmethylene blue assays. Moreover, the expression of CD44 was increased in the presence of both HA and hypoxia compared to either alone. Using a bioinformatic database, hypoxia inducible factor-1α (HIF-1α), a key transcription factor in the hypoxic condition, was found to have 4 predicted binding sites on the CD44 promoter. CD44 expression was significantly increased by treatment with cobalt chloride or dimethyloxalylglycine. Over-expression of HIF-1α in NPCs significantly up-regulated the expression of CD44. The binding site of HIF-1α in the CD44 promoter region, was identified by promoter truncation experiments and chromatin immunoprecipitation assays. Taken together, these results indicated that hypoxic conditions positively potentiated the ability of NPCs matrix synthesis in the presence of HA, which correlated with the increasing CD44 expression by HIF-1α transcriptional activation.