基础医学与临床 ›› 2022, Vol. 42 ›› Issue (6): 913-919.doi: 10.16352/j.issn.1001-6325.2022.06.022

• 研究论文 • 上一篇    下一篇

LncRNA SNHG14调控miR-142-5p对寡聚态Aβ诱导的鼠源神经细胞系PC12损伤的影响

沈杰1, 严洁2, 钟明1, 庞睿娟1, 罗永杰3*   

  1. 成都市第二人民医院 1.神经内科; 2.检验科, 四川 成都 610017;
    3.四川省医学科学院 四川省人民医院 神经内科, 四川 成都 610072
  • 收稿日期:2021-10-28 修回日期:2022-01-28 出版日期:2022-06-05 发布日期:2022-06-02
  • 通讯作者: * m529mo@163.com
  • 基金资助:
    四川省科技厅科研基金重点项目(2019YFS0214);四川省卫计委科研基金重点项目(18ZD011)

Effect of lncRNA SNHG14 on oligomeric Aβ-induced injury in rat neural cell line PC12 through regulating miR-142-5p

SHEN Jie1, YAN Jie2, ZHONG Ming1, PANG Rui-juan1, LUO Yong-jie3*   

  1. 1. Department of Neurology; 2. Department of Laboratory, Chengdu Second People's Hospital, Chengdu 610017;
    3. Department of Neurology, Sichuan Provincial People's Hospital, Sichuan Academy of Medical Sciences, Chengdu 610072,China
  • Received:2021-10-28 Revised:2022-01-28 Online:2022-06-05 Published:2022-06-02
  • Contact: * m529mo@163.com

摘要: 目的 探讨lncRNA SNHG14靶向miR-142-5p对寡聚态Aβ诱导的鼠源神经细胞系PC12损伤的影响。方法 将PC12细胞分为对照组、模型组、模型+si-NC组、模型+si-SNHG14组、模型+miR-NC组、模型+miR-142-5p组、模型+si-SNHG14+anti-miR-NC组、模型+si-SNHG14+anti-miR-142-5p组。RT-qPCR检测SNHG14和miR-142-5p水平。CCK-8和流式细胞术测量细胞增殖抑制率和凋亡率。ELISA检测培养液中TNF-α、IL-6和IL-10水平。双荧素酶报告实验确定SNHG14和miR-142-5p的靶向关系。结果 AD患者血浆中SNHG14呈高表达(P<0.05),miR-142-5p呈低表达(P<0.05)。与对照组比较,模型组细胞SNHG14水平、增殖抑制率、凋亡率显著升高(P<0.05),miR-142-5p水平显著降低(P<0.05),培养液中TNF-α和IL-6水平显著升高(P<0.05),IL-10水平显著降低(P<0.05)。与model+si-NC组比较,model+si-SNHG14组细胞增殖抑制率、凋亡率显著降低(P<0.05),培养液中TNF-α和IL-6水平显著降低(P<0.05),IL-10水平显著升高(P<0.05)。与model+miR-NC组比较,model+miR-142-5p组细胞增殖抑制率、凋亡率显著降低(P<0.05),培养液中TNF-α和IL-6水平显著降低(P<0.05),IL-10水平显著升高(P<0.05)。miR-142-5p是SNHG14的直接靶点。与model+si-SNHG14+ anti-miR-NC组比较,model+ si-SNHG14+anti-miR-142-5p组细胞增殖抑制率、凋亡率显著升高(P<0.05),培养液中TNF-α和IL-6水平显著升高(P<0.05),IL-10水平显著降低(P<0.05)。结论 沉默SNHG14通过靶向促进miR-142-5p表达能够减轻寡聚态Aβ诱导的神经细胞损伤。

关键词: 阿尔茨海默病, SNHG14, miR-142-5p, 寡聚态Aβ, 神经细胞

Abstract: Objective To investigate the effect of lncRNA SNHG14 targeting at miR-142-5p on the neuronal damage induced by oligomeric Aβ. Methods PC12 cells were divided into control group, model group, model+si-NC group, model+si-SNHG14 group, model+miR-NC group, model+miR-142-5p group, model+si-SNHG14+anti-miR-NC group, and model+ si-SNHG14+anti-miR-142-5p group. The level of SNHG14 and miR-142-5p was calculated by RT-qPCR. CCK-8 method and flow cytometry were applied to detect inhibition rate on cell proliferation and apoptosis rate. ELISA method was used to detect TNF-αand IL-6 in the culture. Dual luciferase report assay was used to find the targeting relationship between SNHG14 and miR-142-5p. Results SNHG14 was highly expressed (P<0.05)while miR-142-5p was lowly expressed (P<0.05) in plasma of AD patients. Compared with the control group, the SNHG14 level, cell proliferation inhibition rate, and apoptosis rate in the model group were significantly increased (P<0.05), the level of miR-142-5p was significantly reduced (P<0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly increased (P<0.05), IL-10 level was significantly decreased (P<0.05). Compared with the model+si-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+si-SNHG14 group were significantly reduced (P<0.05), the level of TNF-α and IL-6 in the culture medium was significantly reduced (P<0.05), IL-10 level was significantly increased(P<0.05). Compared with the model+miR-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+miR-142-5p group were significantly reduced (P<0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly reduced (P<0.05) and IL-10 level was significantly increased (P<0.05). miR-142-5p was a direct target of SNHG14. Compared with the model+si-SNHG14+ anti-miR-NC group, the cell proliferation inhibition rate and apoptosis rate in the model+ si-SNHG14+anti-miR-142-5p group were significantly increased(P<0.05), and the levels of TNF-α and IL-6 in the culture medium were significantly increased (P<0.05), IL-10 level was significantly decreased (P<0.05). Conclusions Silencing SNHG14 may alleviate oligomeric Aβ-induced neuronal damage by targeting at and promoting miR-142-5p expression.

Key words: Alzheimer's disease, SNHG14, miR-142-5p, oligomeric Aβ, nerve cell

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