实用医学杂志 ›› 2023, Vol. 39 ›› Issue (17): 2164-2170.doi: 10.3969/j.issn.1006-5725.2023.17.003

• 基础研究 • 上一篇    下一篇

LncRNA NEAT1调控NF-κB信号通路对棕榈酸诱导的LO2细胞损伤与炎症反应的影响

许云春,于馨雅,王雅芝,申元英,郭乐()   

  1. 大理大学基础医学院 (云南 大理 671000 )
  • 收稿日期:2023-03-18 出版日期:2023-09-10 发布日期:2023-09-27
  • 通讯作者: 郭乐 E-mail:le.guo@dali.edu.cn
  • 基金资助:
    国家自然科学基金项目(81960371);云南省科技厅青年项目(202001AU070014);云南省教育厅科学研究基金项目(2023Y0951)

Effect of lncRNA NEAT1 on palmitic acid⁃induced LO2 cell injury and inflammatory response through regulation of NF⁃κB signaling pathway

Yunchun XU,Xinya YU,Yazhi WANG,Yuanying SHEN,Le. GUO()   

  1. School of Basic Medicine,Dali University,Dali 671000,China
  • Received:2023-03-18 Online:2023-09-10 Published:2023-09-27
  • Contact: Le. GUO E-mail:le.guo@dali.edu.cn

摘要:

目的 探讨lncRNA NEAT1调控NF-κB信号通路在棕榈酸诱导的LO2细胞损伤与炎症反应中的作用及机制。 方法 棕榈酸处理LO2细胞24 h;油红O染色和甘油三酯含量检测脂质积累;CCK8检测细胞活力;ALT、AST酶活力检测肝细胞受损情况;RT-qPCR检测lncRNA NEAT1的表达;RT-qPCR和ELISA检测细胞内和培养液上清中炎症因子TNF-α、IL-1β、IL-6的表达;蛋白质印迹和免疫荧光法检测NF-κB信号通路相关蛋白表达。 结果 棕榈酸处理的LO2细胞中,脂质含量增加,细胞活力降低,肝细胞损伤指标ALT、AST酶活性升高,炎症因子TNF-α、IL-1β、IL-6表达增加,同时lncRNA NEAT1表达上调(P < 0.05)。过表达lncRNA NEAT1后,ALT、AST、TNF-α、IL-1β、IL-6表达进一步增加,而细胞活力显著降低,沉默lncRNA NEAT1则起相反作用(P < 0.05)。棕榈酸诱导的LO2细胞中NF-κB信号通路被激活,IKBα蛋白降低,NF-κB p65磷酸化入核增加,过表达lncRNA NEAT1进一步减少IKBα并增加NF-κB p65磷酸化入核,沉默lncRNA NEAT1则起相反作用(P < 0.05)。 结论 棕榈酸诱导的LO2细胞中lncRNA NEAT1表达上调促进细胞损伤与炎症反应,其机制与激活NF-κB信号通路有关。

关键词: lncRNA NEAT1, 代谢相关脂肪性肝病, 肝细胞损伤, 炎症反应, NF-κB信号通路

Abstract:

Objective To explore the role and mechanism of lncRNA NEAT1 in palmitic acid-induced injury and inflammatory response of LO2 cells by regulating NF-κB signaling pathway. Methods LO2 cells were treated with palmitic acid for 24 h. Oil red O staining and triglyceride (TG) test were done to detect lipid accumulation, CCK8 assay was done for cell viability, and ALT and AST were tested for detecting hepatocyte injury. The expression of lncRNA NEAT1 was detected by RT-qPCR, the expression of inflammatory cytokines TNF-α, IL-1β and IL-6 in intracellular and culture supernatants were determined by RT-qPCR and ELISA and the expression of NF-κB signaling pathway-related proteins was detected by Western blot and immunofluorescence. Results In the palmitic acid-treated LO2 cells, the lipid accumulation was increased, the cell viability was decreased, the levels of ALT and AST were increased. Meanwhile, the expression of inflammatory cytokines TNF-α, IL-1β and IL-6 was increased and the expression of lncRNA NEAT1 was upregulated (P < 0.05). After overexpression of lncRNA NEAT1, the levels of ALT, AST, TNF-α, IL-1β, IL-6 were even more increased, the cell viability was significantly reduced. Contrarily, inhibition of lncRNA NEAT1 resulted in the opposite effect (P < 0.05). In addition, NF-κB signaling pathway was activated in the LO2 cells induced by palmitic acid and IKBα expression was decreased and phosphorylation of NF-κB p65 was increased. Then the overexpress of lncRNA NEAT1 further promoted the degradation of IKBα protein and increased NF-κB p65 phosphorylation into the nucleus, and contrarily, inhibition of lncRNA NEAT1 resulted in the opposite effect (P < 0.05). Conclusion The up-expressed lncRNA NEAT1 in LO2 cells induced by palmitic acid can promote cell injury and inflammatory response by way of activating NF-κB signaling pathway.

Key words: lncRNA NEAT1, metabolic associated fatty liver disease, hepatocyte injury, inflammation, NF-κB signaling pathway

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