职业性汞暴露对人肝星状细胞系LX-2纤维化的影响及其机制

职业与健康 ›› 2025, Vol. 41 ›› Issue (11): 1462-1468.

职业性汞暴露对人肝星状细胞系LX-2纤维化的影响及其机制

孙朋朋¹, 周涛²

1.济宁市第一人民医院医学检验科中心实验室,山东济宁 272000;
2.达州市中心医院小儿外科,四川达州 635000

收稿日期:2024-07-15 修回日期:2024-07-29 发布日期:2025-12-12
通信作者: 周涛,主任医师,E-mail：Zhtxn2008@163.com
作者简介:孙朋朋,男,医学中级检验师,主要从事肝纤维化机制的研究工作。
基金资助:
成都2022年度杏林学者学科人才科研提升项目（YYZ X2022181）

Effects and mechanisms of occupational mercury exposure on fibrosis in human hepatic stellate cell line（LX-2）

SUN Pengpeng¹, ZHOU Tao²

1. Central Laboratory, Medical Laboratory Department, Jining First People's Hospital, Jining, Shandong 272000, China;
2. Department of Pediatric Surgery, Dazhou Central Hospital, Dazhou, Sichuan 635000, China

Received:2024-07-15 Revised:2024-07-29 Published:2025-12-12
Contact: ZHOU Tao,Chief physician,E-mail：Zhtxn2008@163.com

摘要/Abstract

摘要： 目的基于小分子核糖核酸-143-3p（microRNA-143-3p,miR-143-3p） -血管生成素样蛋白8（angiopoietin-like protein 8,ANGPTL8）通路探讨职业性汞暴露对人肝星状细胞系（LX-2）纤维化的作用。方法人肝星状细胞 LX-2经胰蛋白酶消化并以4×10³个细胞/孔铺板于96孔板中,孵育24 h后加入不同浓度的HgCl₂（0、2.50、5.00、10.00 μmol/L）,将细胞孵育72 h后分别测定细胞增殖水平、单克隆形成数目、凋亡水平、细胞周期G2/M期;免疫荧光法测定细胞纤维化指标α平滑肌肌动蛋白（α-smooth muscle actin,α-SMA）、Ⅰ型胶原Α1（collagen type Ⅰ α1,Col1a1）;酶联免疫吸附法测定炎症指标肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白细胞介素-6（Interleukin-6,IL-6）、白细胞介素-18（Interleukin-18,IL-18）水平;RT-PCR法及Westen-blot法测定细胞miR-143-3p、ANGPTL8水平。结果与对照组（0 μmol/L HgCl₂）相比,2.50、5.00、10.00 μmol/L HgCl₂暴露剂量组存活率、单克隆形成数目、纤维化指标α-SMA、Col1a1水平均明显升高（均P<0.01）,凋亡率、细胞周期G2/M期水平均明显降低（均P<0.01）,且随着HgCl₂暴露剂量的增加HgCl₂各剂量组存活率、单克隆形成数目、纤维化指标α-SMA、Col1a1水平均逐渐升高（均P<0.01）,凋亡率、细胞周期G2/M期水平均逐渐降低（均P<0.01）。与对照组相比（0.00 μmol/L HgCl₂）,2.50、5.00、10.00 μmol/L HgCl₂暴露剂量组miR-143-3p表达水平（4.83±0.87 vs 2.92±0.62、1.91±0.34、 0.85±0.10）明显降低（P<0.01）;ANGPTL8 mRNA蛋白表达水平（0.95±0.11 vs 1.59±0.16、2.84±0.29、3.69±0.38,0.23±0.06 vs 0.46±0.12、0.83±0.17、1.49±0.21）、TNF-α、IL-6、IL-18水平均明显升高（均P<0.01）;且随着HgCl₂暴露剂量的增加,HgCl₂各剂量组miR-143-3p表达水平（2.92±0.62 vs 1.91±0.34 vs 0.85±0.10）逐渐降低,ANGPTL8 mRNA蛋白（1.59±0.16 vs 2.84±0.29 vs 3.69±0.38,0.46±0.12 vs 0.83±0.17 vs 1.49±0.21）、TNF-α、IL-6、IL-18水平表达水平逐渐升高（均P<0.01）。结论职业性汞暴露可促进人肝星状细胞系（LX-2）异常增殖,诱导细胞纤维化进展,抑制细胞凋亡,诱导细胞周期阻滞。其机制与汞暴露抑制人肝星状细胞系（LX-2）miR-143-3p表达,进而促进ANGPTL8高表达有关。

关键词: 小分子核糖核酸-143-3p, 血管生成素样蛋白8, 汞, 人肝星状细胞系, 纤维化

Abstract: Objective To explore the effect of occupational mercury exposure on fibrosis in human hepatic stellate cell line（LX-2） based on the microRNA-143-3p（miR-143-3p）-angiopoietin-like protein 8（ANGPTL8） pathway. Methods Human hepatic stellate cells LX-2 were digested with trypsin and plated in a 96-well plate at 4×10³ cells/well. After incubation for 24 hours,different concentrations of HgCl₂（0,100,200,400 μmol/L） were added. The cells were incubated for 72 hours,the cell proliferation level,the number of monoclonal formation,the apoptosis level,and the G2/M phase of the cell cycle were measured. The cell fibrosis indicators α-smooth muscle actin（α-SMA） and type collagen A1（Col1a1） were determined by immunofluorescence assay,the levels of inflammatory indicators tumor necrosis factor alpha（TNF-α）,interleukin-6（IL-6）,and interleukin-18（IL-18） were measured by enzyme-linked immunosorbent assay,and the levels of cellular miR-143-3p and ANGPTL8 were measured by RT-PCR and Westen-blot methods. Results Compared with the control group（0.00 μmol/L HgCl₂）,the survival rate,number of monoclonal formations,and levels of fibrosis indicators α-SMA and Col1a1 in the exposure dose groups of 2.50,5.00,and 10.00 μmol/L HgCl₂ were significantly increased（all P<0.01）,while apoptosis rate and cell cycle G2/M phase levels were significantly reduced（all P<0.01）. With the increase of HgCl₂ exposure dose,the survival rate,the number of monoclonal formation,and the levels of fibrosis indicators α-SMA and Col1a1 in each dose group of HgCl₂ gradually increased（all P<0.01）,and the apoptosis rate and cell cycle G2/M phase levels gradually decreased（all P<0.01）. Compared with the control group（0.00 μmol/L HgCl₂）,the expression levels of miR-143-3p in the exposure dose groups of 2.50,5.00 and 10.00 μmol/L HgCl₂（4.83±0.87 vs 2.92±0.62,1.91±0.34,0.85±0.10）were significantly reduced（all P<0.01）,while the expression levels of ANGPTL8 mRNA protein（0.95±0.11 vs 1.59±0.16,2.84±0.29,3.69±0.38;0.23±0.06 vs 0.46±0.12,0.83±0.17,1.49±0.21）,TNF-α,IL-6,and IL-18 increased significantly（all P<0.01）. With the increase in HgCl2 exposure dose,the expression levels of miR-143-3p in each dose group of HgCl₂ gradually decreased（2.92±0.62 vs 1.91±0.34 vs 0.85±0.10）,while the expression levels of ANGPTL8 mRNA protein（1.59±0.16 vs 2.84±0.29 vs 3.69±0.38,0.46±0.12 vs 0.83±0.17 vs 1.49±0.21）,TNF-α,IL-6 and IL-18 gradually increased（all P<0.01）. Conclusion Mercury exposure can promote abnormal proliferation of human hepatic stellate cell line（LX-2）,induce fibrosis progression,inhibit cell apoptosis,and induce cell cycle arrest. The mechanism is related to the inhibition of miR-143-3p expression in human hepatic stellate cell line（LX-2） by mercury exposure,which in turn promotes high expression of ANGPTL8.

Key words: MicroRNA-143-3p, Angiopoietin-like protein 8, Mercury, Human hepatic stellate cell line, Fibrosis

孙朋朋, 周涛. 职业性汞暴露对人肝星状细胞系LX-2纤维化的影响及其机制. [J]职业与健康, 2025, 41(11): 1462-1468.

SUN Pengpeng, ZHOU Tao. Effects and mechanisms of occupational mercury exposure on fibrosis in human hepatic stellate cell line（LX-2）. [J]OCCUPATION AND HEALTH, 2025, 41(11): 1462-1468.

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