lncRNA BDNF-AS 敲低通过下调 miR-130b-5p 靶基因 PRDM5 抑制急性脊髓损伤中的神经元细胞凋亡。
Knockdown of lncRNA BDNF-AS suppresses neuronal cell apoptosis via downregulating miR-130b-5p target gene PRDM5 in acute spinal cord injury.
机构信息
a Department of Orthopedics , The First Affiliated Hospital of Zhengzhou University , Zhengzhou , Henan Province , China.
b Department of Urology , The First Affiliated Hospital of Zhengzhou University , Zhengzhou , Henan Province , China.
出版信息
RNA Biol. 2018;15(8):1071-1080. doi: 10.1080/15476286.2018.1493333. Epub 2018 Sep 10.
OBJECTIVE
The present study was designed to investigate the molecular mechanism and biological roles of lncRNA brain-derived neurotrophic factor antisense (lncRNA BDNF-AS) in acute spinal cord injury (ASCI).
METHODS
The rat model of ASCI and hypoxic cellular model were established to detect the expression of BDNF-AS, miR-130b-5p, PR (PRDI-BF1 and RIZ) domain protein 5 (PRDM5) and cleaved caspase 3 (c-caspase 3) using qRT-PCR and western blot. Basso, Beattie and Bresnahan (BBB) score was carried out to assess neurological function. Flow cytometry was used to determine the apoptosis of neuronal cells. The association among BDNF-AS, miR-130b-5p and PRDM5 were disclosed by RNA immunoprecipitation (RIP) assay, RNA pull-down assay and dual-luciferase reporter assay.
RESULTS
BDNF-AS, PRDM5 and c-caspase 3 expression were significantly upregulated, while miR-130b-5p was suppressed in the ASCI group and neuronal cells following hypoxia treatment. BDNF-AS knockdown inhibited neuronal cell apoptosis. Further studies indicated that BDNF-AS functioned as a competing endogenous RNA (ceRNA) by sponging miR-130b-5p in neuronal cells. Further investigations demonstrated that PRDM5 was a target of miR-130b-5p and BDNF-AS knockdown exerted anti-apoptotic effects via miR-130b-5p/PRDM5 axis.
CONCLUSION
The lncRNA BDNF-AS/miR-130b-5p/PRDM5 axis might be a promising therapeutic target for ASCI.
目的
本研究旨在探讨长链非编码 RNA 脑源性神经营养因子反义（lncRNABDNF-AS）在急性脊髓损伤（ASCI）中的分子机制和生物学作用。
方法
建立大鼠 ASCI 模型和低氧细胞模型，采用 qRT-PCR 和 Western blot 检测 BDNF-AS、miR-130b-5p、PR（PRDI-BF1 和 RIZ）结构域蛋白 5（PRDM5）和裂解半胱氨酸天冬氨酸蛋白酶 3（c-caspase 3）的表达。采用 Basso、Beattie 和 Bresnahan（BBB）评分评估神经功能。采用流式细胞术检测神经元细胞凋亡。通过 RNA 免疫沉淀（RIP）试验、RNA 下拉试验和双荧光素酶报告基因试验揭示 BDNF-AS、miR-130b-5p 和 PRDM5 之间的关联。
结果
ASCI 组和低氧处理后的神经元细胞中，BDNF-AS、PRDM5 和 c-caspase 3 的表达明显上调，而 miR-130b-5p 的表达受到抑制。BDNF-AS 敲低抑制神经元细胞凋亡。进一步的研究表明，BDNF-AS 在神经元细胞中作为竞争性内源性 RNA（ceRNA）通过海绵吸附 miR-130b-5p 发挥作用。进一步的研究表明，PRDM5 是 miR-130b-5p 的靶基因，BDNF-AS 敲低通过 miR-130b-5p/PRDM5 轴发挥抗凋亡作用。
结论
lncRNA BDNF-AS/miR-130b-5p/PRDM5 轴可能是 ASCI 的一个有前途的治疗靶点。
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