Suppression of miR-203-3p inhibits lipopolysaccharide induced human intervertebral disc inflammation and degeneration through upregulating estrogen receptor α
Suppression of miR-203-3p inhibits lipopolysaccharide induced human intervertebral disc inflammation and degeneration through upregulating estrogen receptor α
复制标题
DOI：
10.1038/s41434-019-0118-z
复制
发表时间：
2020-01-02
期刊：
影响因子：
5.1
通讯作者：
Xu, Hui
中科院分区：
文献类型：
--
作者：
Cai, Zhongxu;Li, Kunpeng;Xu, Hui
越来越多的证据表明，雌激素受体α(ERα)和microRNAs(MiRNAs)在椎间盘退行性变(IDD)中起着关键作用。然而，在IDD发生过程中与ERα相关的特定miRNA仍不清楚。因此，我们旨在探讨ERα相关miRNA在IDD模型中的作用。分离IDD患者的髓核(NP)细胞。用定量逆转录聚合酶链式反应(qRT-PCR)从IDD患者的NP组织中筛选和验证ERα相关miRNAs。并用定量逆转录聚合酶链式反应(qRT-PCR)检测相关细胞因子的mRNA水平。用Western印迹法检测蛋白质水平。采用酶联免疫吸附试验检测培养上清液中炎性细胞因子的浓度。与低级别IDD患者相比，高级别IDD患者NP组织中MIR-203-3p表达上调，且与ERα表达呈负相关。MiR-203-3p直接靶向IDD患者NP细胞中的ERα。内毒素刺激后，NP细胞miR-203-3p表达增加，ERα表达降低。MIR-203-3p抑制可抑制内毒素对ERα表达和IDD相关基因的影响，而ERα下调则可挽救内毒素的作用。结论：抑制miR-203-3p的表达可能通过上调ERα来抑制内毒素诱导的人腰椎间盘的炎症和退变。
Accumulating evidence demonstrates that estrogen receptor alpha (ER alpha) and microRNAs (miRNAs) play crucial roles in intervertebral disc degeneration (IDD). However, the specific miRNA that related with ER alpha during IDD development remains unknown. Therefore, we aimed to explore the role of ER alpha-related miRNA in the IDD model. Nucleus pulposus (NP) cells were isolated from IDD patients. ER alpha-related miRNAs were selected and verified in NP tissues from IDD patients using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Also, the related cytokine mRNA levels were detected by qRT-PCR. Protein levels were determined by Western blot. The concentrations of inflammatory cytokines in culture supernatants were detected by enzyme-linked immunosorbent assay. MiR-203-3p was found to be upregulated in NP tissues of high-grade IDD patients compared with low-grade IDD patients, and negatively associated with ER alpha expression. MiR-203-3p directly targeted ER alpha in NP cells of IDD patients. After lipopolysaccharides (LPS) stimulation, miR-203-3p expression increased, while ER alpha expression decreased in NP cells. MiR-203-3p inhibition suppressed the effect of LPS on ER alpha expression and IDD related genes, while ER alpha downregulation rescued the effect of LPS. In conclusion, suppression the expression of miR-203-3p could inhibit LPS-induced human intervertebral disc inflammation and degeneration through upregulating ER alpha.