MicroRNA-323 suppresses nerve cell toxicity in cerebral infarction via the transforming growth factor-1/SMAD3 signaling pathway
MicroRNA-323 suppresses nerve cell toxicity in cerebral infarction via the transforming growth factor-1/SMAD3 signaling pathway
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DOI：
10.3892/ijmm.2018.4020
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发表时间：
2019-02-01
影响因子：
5.4
通讯作者：
Tong, Yanna
中科院分区：
文献类型：
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作者：
Che, Fengli;Du, Huishan;Tong, Yanna
本研究旨在探讨microRNA-323（miR-323）在脑梗死中的作用及其机制。建立大鼠脑梗死模型，并对海马组织进行分析。此外，为了进一步理解miR-323的作用，用miR-323模拟物或抑制剂转染PC 12细胞，并使其经受缺氧以模拟脑梗死。采用逆转录-定量聚合酶链反应检测miR-323的表达。采用荧光素酶报告基因检测技术，对SMAD 3 mRNA的3 '端非翻译区部分序列进行体外分析。Western印迹分析也用于分析转化生长因子-1（TGF-1）和SMAD 3蛋白表达水平。与假手术对照组相比，脑梗死大鼠中miR-323的表达显著上调。此外，在体外脑梗死模型中，miR-323的过表达诱导神经细胞毒性并减少神经细胞生长，而miR-323的下调在该模型中对神经细胞毒性和生长产生相反的作用。此外，在体外脑梗死模型中，过表达miR-323直接靶向并抑制SMAD 3表达，而抑制miR-323则诱导SMAD 3表达。SMAD 3抑制剂的使用抑制了抗miR-323对脑梗死体外模型中神经细胞毒性的影响。总之，这些发现表明miR-323通过TGF-1/SMAD 3信号通路抑制脑梗死中的神经细胞凋亡。
In the present study, the aim was to investigate the function of microRNA-323 (miR-323) in cerebral infarction and its underlying mechanism. A rat model of cerebral infarction was established and hippocampal tissues were analyzed. In addition, to further understand the role of miR-323, PC12 cells were transfected with miR-323 mimics or inhibitors and subjected to hypoxia to model cerebral infarction. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of miR-323. A luciferase reporter assay was conducted to analyze miR-323 target sites the partial sequence of the 3'-untranslated region of SMAD3 mRNA in vitro. Western blot analysis was also used to analyze transforming growth factor-1 (TGF-1) and SMAD3 protein expression levels. It was observed that miR-323 expression was significantly upregulated in rats with cerebral infarction compared with rats in the sham-control group. In addition, overexpression of miR-323 induced nerve cell toxicity and reduced nerve cell growth in an in vitro model of cerebral infarction, whereas downregulation of miR-323 caused the opposite effects on nerve cell toxicity and growth in this model. In addition, overexpression of miR-323 directly targeted and suppressed SMAD3 expression in the in vitro model of cerebral infarction, while inhibition of miR-323 induced SMAD3 expression. The use of a SMAD3 inhibitor suppressed the effect of anti-miR-323 on nerve cell toxicity in the in vitro model of cerebral infarction. Collectively, these findings suggested that miR-323 suppresses nerve cell apoptosis in cerebral infarction via the TGF-1/SMAD3 signaling pathway.