Mechanism of lncRNA NEAT1 regulating chondrocyte pyroptosis in osteoarthritis

TANG Jie; ZHANG Pi-jun; LIN Wei; LI Chao-yi

doi:10.11659/jjssx.03E024082

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Mechanism of lncRNA NEAT1 regulating chondrocyte pyroptosis in osteoarthritis

Basic Research | 更新时间：2025-03-25

- Mechanism of lncRNA NEAT1 regulating chondrocyte pyroptosis in osteoarthritis
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 34, Issue 3, Pages: 199-204(2025)
- 作者机构：
  
  海南医科大学第二附属医院关节外科，海南海口 570311
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.03E024082
  CLC： R684.3
- Received：15 March 2024，
  
  Published：25 March 2025
- 稿件说明：
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唐捷,张丕军,林维等.lncRNA NEAT1调控骨关节炎软骨细胞焦亡的机制[J].局解手术学杂志,2025,34(03):199-204.

TANG Jie,ZHANG Pi-jun,LIN Wei,et al.Mechanism of lncRNA NEAT1 regulating chondrocyte pyroptosis in osteoarthritis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(03):199-204.
唐捷,张丕军,林维等.lncRNA NEAT1调控骨关节炎软骨细胞焦亡的机制[J].局解手术学杂志,2025,34(03):199-204. DOI： 10.11659/jjssx.03E024082.

TANG Jie,ZHANG Pi-jun,LIN Wei,et al.Mechanism of lncRNA NEAT1 regulating chondrocyte pyroptosis in osteoarthritis[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(03):199-204. DOI： 10.11659/jjssx.03E024082.

摘要

目的

探讨长链非编码RNA（lncRNA）核富集丰富转录本1（NEAT1）对骨关节炎（OA）软骨细胞焦亡信号轴的影响和潜在机制。

方法

收集6例OA患者的膝关节软骨组织和6例正常软骨组织。使用TargetScan 7.2和StarBase预测miR-26a与lncRNA NEAT1的靶向关系，双荧光素酶报告基因实验验证lncRNA NEAT1与miR-26a的靶向调控作用。将培养的软骨细胞系分为对照组（不进行药物干预）、IL-1β组（添加10 μg/L IL-1β诱导体外OA）、IL-1β+NEAT1-EV组（添加10 μg/L IL-1β，联合转染20 μg/L的NEAT1-EV）、IL-1β+NEAT1-OE组（添加10 μg/L IL-1β，联合转染20 μg/L的NEAT1-OE）、IL-1β+NEAT1-OE+mimic-NC组（添加10 μg/L IL-1β，联合转染20 μg/L的NEAT1-OE和50 μg/L的mimic-NC）、IL-1β+NEAT1-OE+mimic组（添加10 μg/L IL-1β，联合转染20 μg/L的NEAT1-OE和50 μg/L的miR-26a mimic）。qRT-PCR法检测OA软骨组织和细胞中lncRNA NEAT1和miR-26a的表达水平。CCK-8法检测各组细胞的活力，蛋白免疫印迹法检测各组Caspase1和cleaved-Caspase1蛋白表达及细胞焦亡标志物[NOD样受体蛋白3（NLRP3）、白细胞介素18（IL-18）和切割模式的Gasdermin D（cleaved-Gasdermin D）]的表达。

结果

OA软骨组织中lncRNA NEAT1的表达水平高于正常软骨组织（

0.05），而miR-26a的表达水平低于正常软骨组织（

0.05）。双荧光素酶报告基因实验证实，lncRNA NEAT1与miR-26a具有靶向调控作用。与对照组比较，IL-1β组细胞中lncRNA NEAT1、cleaved-Caspase1、NLRP3、IL-18、cleaved-Gasdermin D的表达水平均上调（

0.05），而miR-26a表达水平下调（

0.05），细胞活力降低（

0.05）。与IL-1β+NEAT1-EV组比较，IL-1β+NEAT1-OE组细胞中lncRNA NEAT1、cleaved-Caspase1、NLRP3、IL-18、cleaved-Gasdermin D的表达水平均上调（

0.05），而miR-26a表达水平下调（

0.05），细胞活力下降（

0.05）。与IL-1β+NEAT1-OE+mimic-NC组比较，IL-1β+NEAT1-OE+mimic组细胞中cleaved-Caspase1、NLRP3、IL-18、cleaved-Gasdermin D的表达水平均下调（

0.05），而miR-26a表达水平上调（

0.05），细胞活力升高（

0.05）。

结论

lncRNA NEAT1通过靶向抑制miR-26a激活OA软骨细胞中Caspase1介导的焦亡信号轴。

Abstract

Objective

To investigate the effect of long non-coding RNA （lncRNA） nuclear enriched abundant transcript 1 （NEAT1） on the chondrocyte pyroptosis signaling axis in osteoarthritis （OA） and its potential mechanism.

Methods

Knee joint cartilage tissues were collected from 6 OA patients and 6 healthy individuals. The targeting relationship between miR-26a and lncRNA NEAT1 was predicted by TargetScan 7.2 and StarBase，and the targeting regulatory effect of lncRNA NEAT1 and miR-26a was verified by dual luciferase reporter assays. The cultured chondrocyte lines were divided into the control group （without drug intervention）， IL-1β group （with 10 μg/L of IL-1β for inducing OA in vitro），IL-1β+NEAT1-EV group （with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-EV），IL-1β+NEAT1-OE group （with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-OE）， IL-1β+NEAT1-OE+mimic-NC （with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of mimic-NC）， and IL-1β+NEAT1-OE+mimic （with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of miR-26a mimic）. The expression levels of lncRNA NEAT1 and miR-26a in OA cartilage tissues and cells were detected using qRT-PCR. The cell viability was determined using CCK-8 assay. The expression of Caspase1 and cleaved-Caspase1， as well as the pyroptosis markers ［NOD-like receptor family pyrin domain containing 3 （NLRP3）， interleukin-18 （IL-18） and cleaved-Gasdermin D］ were detected using Western blot.

Results

In the OA cartilage tissue， the expression level of lncRNA NEAT1 was higher than that in the normal cartilage tissue， while the expression level of miR-26a was lower than that in the normal cartilage group （

0.05）. The dual luciferase reporter assays confirmed the targeted regulatory effect of lncRNA NEAT1 and miR-26a. Compared with the control group， the IL-1β group showed upregulation of lncRNA NEAT1， cleaved-Caspase1， NLRP3

， IL-18， and cleaved-Gasdermin D expression levels （

0.05），downregulation of miR-26a expression level （

0.05），and a decrease of cell viability（

0.05）.Compared with the IL-1β+NEAT1-EV group， the IL-1β+NEAT1-OE group exhibited upregulation of lncRNA NEAT1， cleaved-Caspase1， NLRP3， IL-18， and cleaved-Gasdermin D expression levels （

0.05）， downregulation of miR-26a expression level （

0.05）， and a decrease of cell viability （

0.05）. Compared with the IL-1β+NEAT1-OE+mimic-NC group， the IL-1β+NEAT1-OE+mimic group showed downregulation of cleaved-Caspase1， NLRP3， IL-18， and cleaved-Gasdermin D expression levels， upregulation of miR-26a expression level （

0.05）， and an increase of cell viability （

0.05）.

Conclusion

lncRNA NEAT1 activates the chondrocyte pyroptosis signaling axis mediated by Caspase1 in OA by the targeted inhibition of miR-26a.

关键词

Keywords

references

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