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长链非编码 RNA GAS5 通过调控 miR-433-3p/TLR10 轴抑制 NF-κB 信号通路抑制 LX-2 细胞的活化。

Long noncoding RNA GAS5 inhibits LX-2 cells activation by suppressing NF-κB signalling through regulation of the miR-433-3p/TLR10 axis.

机构信息

Department of Gastroenterology, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Province, P.R. China.

Department of Gastroenterology, First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Province, P.R. China.

出版信息

Dig Liver Dis. 2022 Aug;54(8):1066-1075. doi: 10.1016/j.dld.2021.11.002. Epub 2021 Dec 10.

Abstract

BACKGROUND

Liver fibrosis is a common disease that can lead to hepatic failure.

AIMS

Our aims were to reveal the role of GAS5 in the regulation of liver fibrosis.

METHODS

LX-2 human hepatic satellite cells (HSCs) were cultured and activated using TGF-β1 treatment. A CCK-8 assay was performed to assess cell viability. A luciferase assay was employed to monitor the interactions between miR-433-3p and GAS5 or toll-like receptor 10 (TLR10). Western blotting and real-time quantitative PCR (RT-qPCR) were applied to detect the expression levels of α-SMA, Col. I, PCNA-, MMP2-, MMP9-, TLR10-, and NF-κB-related molecules at the protein and RNA levels.

RESULTS

GAS5 and TLR10 were decreased while miR-433-3p was upregulated in TGF-β1-activated LX-2 cells. Upregulation of GAS5 or downregulation of miR-433-3p suppressed HSC activation, and luciferase assays indicated that miR-433-3p binds with GAS5 and the 3'-UTR of TLR10. MiR-433-3p upregulation and TLR10 downregulation rescued the impacts of GAS5 overexpression or miR-433-3p knockdown on LX-2 cells. Upregulation of GAS5 also suppressed the phosphorylation of NF-κB through the miR-433-3p/TLR10 axis.

CONCLUSION

LncRNA GAS5 exerts an inhibitory effect on HSC activation by suppressing NF-κB signalling through regulation of the miR-433-3p/TLR10 axis.

摘要

背景

肝纤维化是一种常见疾病,可导致肝衰竭。

目的

本研究旨在揭示 GAS5 在肝纤维化调控中的作用。

方法

采用 TGF-β1 处理培养和激活 LX-2 人肝星状细胞(HSCs)。用 CCK-8 法检测细胞活力。采用荧光素酶报告基因实验检测 miR-433-3p 与 GAS5 或 Toll 样受体 10(TLR10)之间的相互作用。采用 Western blot 和实时定量 PCR(RT-qPCR)检测α-SMA、Col. I、PCNA-、MMP2-、MMP9-、TLR10-和 NF-κB 相关分子在蛋白和 RNA 水平上的表达。

结果

在 TGF-β1 激活的 LX-2 细胞中,GAS5 和 TLR10 表达下调,而 miR-433-3p 表达上调。GAS5 上调或 miR-433-3p 下调抑制 HSC 激活,荧光素酶报告基因实验表明 miR-433-3p 与 GAS5 和 TLR10 的 3'-UTR 结合。miR-433-3p 上调和 TLR10 下调挽救了 GAS5 过表达或 miR-433-3p 敲低对 LX-2 细胞的影响。GAS5 上调还通过 miR-433-3p/TLR10 轴抑制 NF-κB 的磷酸化。

结论

lncRNA GAS5 通过调控 miR-433-3p/TLR10 轴抑制 NF-κB 信号通路,对 HSC 激活发挥抑制作用。

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