长链非编码 RNA BDNF-AS 通过消除 microRNA-125b-5p 促进 MPTP 诱导的帕金森病中的自噬和细胞凋亡。
LncRNA BDNF-AS promotes autophagy and apoptosis in MPTP-induced Parkinson's disease via ablating microRNA-125b-5p.
机构信息
Department of Neurology, Liaocheng People's Hospital, No. 67, West Dongchang Road, Liaocheng City, Shandong Province, 252000, China.
Department of Neurology, Liaocheng Third People's Hospital, No. 62, Weiyu Road, Liaocheng City, Shandong Province, 252000, China.
出版信息
Brain Res Bull. 2020 Apr;157:119-127. doi: 10.1016/j.brainresbull.2020.02.003. Epub 2020 Feb 10.
BACKGROUNDS
Recently, extensive evidence has indicated that the biological role of long non-coding RNAs (lncRNAs) in neurodegenerative diseases is becoming increasingly evident. The lncRNA brain-derived neurotrophic factor anti-sense (BDNF-AS) has been found to be dysregulated in Huntington's Disease. However, the function of BDNF-AS in Parkinson's disease (PD) remains unknown. The purpose of this present study was to explore the effect of BDNF-AS on PD and its underlying molecular mechanisms.
METHODS
The MPTP-induced mouse model of PD and MPP+-induced SH-SY5Y cell model were established. Immunofluorescence was performed to determine the number of TH + positive cells. Mice behavioral changes were detected by pole and rota-rod test. SH-SY5Y cells viability, apoptosis was detected by MTT assay and flow cytometry. The number of autophagosome was measured by transmission electron microscopy. Dopamine content was tested by high performance liquid chromatography. Dual-luciferase reporter gene assay was utilized to verify the correlation between BDNF-AS and miR-125b-5p. qRT-PCR and western blot were used to detect gene expression levels.
RESULTS
Our results showed that BDNF-AS was up-regulated in MPTP-induced PD model and dopamine neurons, and MPP + treated SH-SY5Y cells, while miR-125b-5p was down-regulated. The expression of BDNF-AS was positively related with the MPP + concentration. BDNF-AS knockdown could significantly promote cell proliferation, while inhibit apoptosis and autophagy in SH-SY5Y cells treated by MPP + . Silencing BDNF-AS could also increase TH positive neurons and significantly suppress the autophagy of PD mice. Additionally, miR-125b-5p, a putative target gene of BDNF-AS, was involved in the effects of BDNF-AS on SH-SY5Y cell apoptosis and autophagy.
CONCLUSIONS
Our study demonstrated that knockdown of BDNF-AS could elevate SH-SY5Y cell viability, inhibit autophagy and apoptosis in MPTP-induced PD models through regulating miR-125b-5p, suggesting that BDNF-AS might act as a potential therapeutic target for PD.
背景
最近，大量证据表明长非编码 RNA（lncRNA）在神经退行性疾病中的生物学作用越来越明显。研究发现脑源性神经营养因子反义（BDNF-AS）在亨廷顿病中失调。然而，BDNF-AS 在帕金森病（PD）中的作用尚不清楚。本研究旨在探讨 BDNF-AS 对 PD 的影响及其潜在的分子机制。
方法
建立 MPTP 诱导的 PD 小鼠模型和 MPP+诱导的 SH-SY5Y 细胞模型。免疫荧光法测定 TH+阳性细胞数。通过杆和转棒试验检测小鼠行为变化。MTT 法和流式细胞术检测 SH-SY5Y 细胞活力和凋亡。透射电镜测量自噬体数量。高效液相色谱法检测多巴胺含量。双荧光素酶报告基因实验验证 BDNF-AS 与 miR-125b-5p 的相关性。qRT-PCR 和 Western blot 检测基因表达水平。
结果
我们的结果表明，BDNF-AS 在 MPTP 诱导的 PD 模型和多巴胺神经元中上调，在 MPP+处理的 SH-SY5Y 细胞中也上调，而 miR-125b-5p 下调。BDNF-AS 的表达与 MPP+浓度呈正相关。BDNF-AS 敲低可显著促进 MPP+处理的 SH-SY5Y 细胞增殖，同时抑制细胞凋亡和自噬。沉默 BDNF-AS 还可以增加 PD 小鼠中 TH 阳性神经元，并显著抑制 PD 小鼠的自噬。此外，BDNF-AS 的潜在靶基因 miR-125b-5p 参与了 BDNF-AS 对 SH-SY5Y 细胞凋亡和自噬的影响。
结论
本研究表明，BDNF-AS 敲低可通过调节 miR-125b-5p 提高 MPTP 诱导的 PD 模型中 SH-SY5Y 细胞的活力，抑制自噬和凋亡，提示 BDNF-AS 可能成为 PD 的潜在治疗靶点。
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