摘 要：为评估长链非编码RNA（lncRNA）lnc-ISG20对转化生长因子β1（TGF-β1）诱导的肝星状细胞活化的作用，探究沉默lnc-ISG20抗肝纤维化的作用机制，本研究用GEO数据库分析lnc-ISG20在肝纤维化组织和正常肝组织中的表达差异。采用10 ng/mL的TGF-β1处理人肝星状细胞LX-2，建立肝纤维化细胞模型，并将其分为sh-NC组和sh-lnc-ISG20组，分别转染sh-NC质粒和sh-lnc-ISG20质粒。利用CCK-8试验检测各组LX-2细胞的生长曲线；利用流式细胞术检测各组LX-2细胞的周期分布和凋亡率；利用双荧光素酶报告试验验证lnc-ISG20与miR-20a-5p的靶向关系；利用GEO数据库分析lnc-ISG20与miR-20a-5p在肝纤维化组织中表达的相关性；利用qRT-PCR检测各组细胞中miR-20a-5p的表达水平；利用Western blotting检测MAPK/NF-κB信号通路蛋白p-P38、p-IκB、p-P65、p-ASK1的表达水平。试验结果表明：与正常肝组织相比，肝纤维化组织中lnc-ISG20表达升高（P<0.01）；与sh-NC组相比，sh-lnc-ISG20组LX-2细胞增殖活力降低（P<0.05），细胞周期被阻滞（P<0.01），细胞凋亡率增加（P<0.01）；miR-20a-5p是lnc-ISG20的靶基因（P<0.01）；肝纤维化组织中lnc-ISG20与miR-20a-5p表达呈负相关（P<0.01）；与sh-NC组相比，sh-lnc-ISG20组LX-2细胞miR-20a-5p表达升高（P<0.01），MAPK/NF-κB信号通路蛋白p-P38、p-IκB、p-P65、p-ASK1表达均降低（均P<0.01）；沉默lnc-ISG20通过升高miR-20a-5p表达，抑制TGF-β1诱导的LX-2细胞的增殖并促进细胞凋亡。本研究表明，lnc-ISG20有望成为肝纤维化治疗的分子靶标。

关键词：肝纤维化；lnc-ISG20；miR-20a-5p；肝星状细胞；细胞凋亡
中图分类号：R575                            文献标志码：A            DOI：10.3969/j.issn.1007-7146.2025.05.008

Abstract: To evaluate the effect of long noncoding RNA (lncRNA) lnc-ISG20 on transforming growth factor β1(TGF-β1)-induced hepatic stellate cell activation and explore the mechanism of silencing lnc-ISG20 in anti-liver fibrosis. The GEO database was used to analyze the differential expression of lnc-ISG20 in liver fibrosis tissue and normal liver tissue. The liver fibrosis cell model was established by treating human hepatic stellate cells LX-2 with 10 ng/mL TGF-β1 and then divided into sh-NC group and sh-lnc-ISG20 group, which were transfected with sh-NC plasmid and sh-lnc-ISG20 plasmid, respectively. CCK-8 assay was used to detect the growth curve of LX-2 cells in each group. Flow cytometry was used to detect the cell cycle distribution and apoptosis rate of LX-2 cells in each group. Dual luciferase reporter assay was used to verify the targeting relationship between lnc-ISG20 and miR-20a-5p. The GEO database was used to analyze the correlation between the expression of lnc-ISG20 and miR-20a-5p in liver fibrosis tissues. qRT-PCR was used to detect the expression level of miR-20a-5p in each group of cells. Western blotting was used to detect the expression levels of p-P38, p-IκB, p-P65 and p-ASK1 proteins in the MAPK/NF-κB signaling pathway. Compared with normal liver tissue, the expression of lnc-ISG20 in liver fibrosis tissue increased (P<0.01). Compared with the sh-NC group, the proliferation activity of LX-2 cells in the sh-lnc-ISG20 group decreased (P<0.05), the cell cycle was blocked (P<0.01), and the apoptosis rate increased (P<0.01). miR-20a-5p was a target gene of lnc-ISG20 (P<0.01). The expression of lnc-ISG20 is negatively correlated with miR-20a-5p in liver fibrosis tissue (P<0.01). Compared with the sh-NC group, the expression of miR-20a-5p in LX-2 cells in the sh-lnc-ISG20 group increased (P<0.01), and the expressions of p-P38, p-IκB, p-P65 and p-ASK1 proteins of the MAPK/NF-κB signaling pathway all decreased (all P<0.01). Silencing lnc-ISG20 inhibits TGF-β1-induced proliferation of LX-2 cells and promotes apoptosis by increasing miR-20a-5p expression. lnc-ISG20 is expected to become a molecular target for the treatment of liver fibrosis.
Key words: liver fibrosis; lnc-ISG20; miR-20a-5p; hepatic stellate cells; cell apoptosis
（Acta Laser Biology Sinica, 2025, 34(5): 451-458）