中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (24): 6259-6266.doi: 10.12307/2026.198

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

孕烷X受体在亚砷酸钠致人正常肝细胞氧化应激及炎症损伤中的作用

张小旭 1，田侦丽2，谢婷婷 1   

  1. 1贵州医科大学附属医院临床检验中心，贵州医科大学医学检验学院，贵州省贵阳市   550004；2六盘水市妇幼保健院，贵州省六盘水市   553001
  • 收稿日期:2025-06-16 修回日期:2025-09-11 出版日期:2026-08-28 发布日期:2026-02-02
  • 通讯作者: 谢婷婷，博士，副教授，副主任技师，贵州医科大学附属医院临床检验中心，贵州医科大学医学检验学院，贵州省贵阳市 550004
  • 作者简介:张小旭，女，2001年生，贵州省遵义市人，汉族，贵州医科大学在读硕士，检验技师，主要从事砷中毒机制研究。
  • 基金资助:
    国家自然科学基金项目(81560514)，项目负责人：谢婷婷；贵州医科大学附属医院2022年博士科研启动基金项目(gyfybsky-2022-33)，项目负责人：谢婷婷

Roles of pregnane X receptor in sodium arsenite-induced oxidative stress and inflammatory injury in human normal hepatocytes

Zhang Xiaoxu1, Tian Zhenli2, Xie Tingting1   

  1. 1Center for Clinical Laboratory, Affiliated Hospital of Guizhou Medical University, School of Clinical Laboratory Science of Guizhou Medical University, Guiyang 550004, Guizhou Province, China; 2Liupanshui Maternal and Child Health Hospital, Liupanshui 553001, Guizhou Province, China
  • Received:2025-06-16 Revised:2025-09-11 Online:2026-08-28 Published:2026-02-02
  • Contact: Xie Tingting, MD, Associate professor, Associate chief technician, Center for Clinical Laboratory, Affiliated Hospital of Guizhou Medical University, School of Clinical Laboratory Science of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • About author:Zhang Xiaoxu, MS candidate, Junior technician, Center for Clinical Laboratory, Affiliated Hospital of Guizhou Medical University, School of Clinical Laboratory Science of Guizhou Medical University, Guiyang 550004, Guizhou Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81560514 (to XTT); 2022 Doctoral Research Initiation Fund Project of Affiliated Hospital of Guizhou Medical University, No. gyfybsky-2022-33 (to XTT)

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摘要：



文题释义：
亚砷酸钠：砷是大气环境中常见的一种类金属，而过量的砷在人体积聚会危害人体健康。亚砷酸钠是砷的一种化合物，为建立砷暴露模型常用的化合物。
孕烷X受体：是参与外源性物质和内生素解毒调节的核受体超家族成员，以配体依赖性方式与靶基因序列结合来调节下游基因表达，影响解毒、代谢、炎症抑制、细胞凋亡、肿瘤迁移和抗氧化应激等各种生物过程。

背景：肝脏作为砷在体内代谢的主要器官，成为砷毒作用机制相关研究的焦点。
目的：研究孕烷X受体在亚砷酸钠致人正常肝细胞氧化应激及炎症损伤中的作用。
方法：将对数期生长状态良好的人正常肝细胞MIHA分4组培养，分别加入0(对照)，10，20，30 μmol/L亚砷酸钠培养48 h，观察细胞形态变化，CCK-8 法检测细胞活力，荧光探针染色结合酶标仪法检测细胞内活性氧水平，硫代巴比妥酸法检测细胞内丙二醛水平，烟酰胺腺嘌呤二核苷酸磷酸氢法检测细胞内谷胱甘肽还原酶活性，WST-8法检测细胞内总超氧化物歧化酶活性，ELISA检测细胞上清中白细胞介素6、白细胞介素1β、肿瘤坏死因子α水平，qRT-PCR检测各组细胞内孕烷X受体、细胞色素P450 3A4酶mRNA表达，Western blot检测孕烷X受体、细胞色素P450 3A4酶、核因子κB p65、核因子κB p-p65、增殖核抗原、白细胞介素6、白细胞介素1β、肿瘤坏死因子α、核因子κB抑制蛋白α、环氧合酶2、p-核因子κB抑制蛋白α、核转录因子红系2相关因子2、Keap1抗体、p-核转录因子红系2相关因子2蛋白表达。
结果与结论：①与对照组相比，各浓度亚砷酸钠组细胞胞膜边界不清，胞质减少，细胞融合率降低，间隙增宽。与对照组相比，各浓度亚砷酸钠组细胞内活性氧、丙二醛水平均升高(P < 0.05)，细胞上清中白细胞介素6、白细胞介素1β、肿瘤坏死因子α水平均升高(P < 0.05)，p-核因子κB抑制蛋白α、核因子κB p-p65、核因子κB p65、肿瘤坏死因子α、白细胞介素1β蛋白表达均升高(P < 0.05)，细胞存活率均降低     (P < 0.05)，增殖核抗原、核转录因子红系2相关因子2、p-核转录因子红系2相关因子2蛋白表达与总超氧化物歧化酶活性均降低(P < 0.05)，孕烷X受体、细胞色素P450 3A4酶mRNA与蛋白表达均降低(P < 0.05)。与对照组相比，20，30 μmol/L亚砷酸钠组细胞内谷胱甘肽还原酶活性降低(P < 0.05)，Keap1、白细胞介素6、环氧合酶2蛋白表达均升高(P < 0.05)。②结果表明，亚砷酸钠可能通过下调孕烷X受体表达抑制转录因子红系2相关因子2抗氧化通路与激活核因子κB炎症通路诱导肝细胞的氧化应激与炎症损伤，同时抑制药物代谢酶细胞色素P450 3A4 酶表达。
https://orcid.org/0009-0000-0920-4737 (张小旭) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 亚砷酸钠, 肝损伤, 孕烷X受体, 氧化应激, 核因子κB信号通路, 炎症反应, CYP3A, 4；组织构建

Abstract: BACKGROUND: As the primary organ for arsenic metabolism in the body, the liver has become a focal point for research on the mechanisms of arsenic toxicity.
OBJECTIVE: To investigate the role of pregnane X receptor in sodium arsenite-induced oxidative stress and inflammatory injury in human normal hepatocytes.
METHODS: Human normal hepatocyte MIHA cells were exposed to 0 (control), 10, 20, 30 μmol/L sodium arsenite for 48 hours. Changes in cell morphology were observed. Cell viability was measured via the cell counting kit-8 assay. Intracellular reactive oxygen species levels were detected using fluorescent probe staining combined with a microplate reader. Intracellular malondialdehyde content, glutathione reductase activity, and total superoxide dismutase activity were assessed using the thiobarbituric acid method, nicotinamide adenine dinucleotide phosphate method, and WST-8 method, respectively. Levels of interleukin-6, tumor necrosis factor-α, and interleukin-1β in the cell supernatant were quantified via ELISA. mRNA expression of pregnane X receptor and cytochrome P450 3A4 enzyme was analyzed using real-time quantitative PCR. Protein expression levels of pregnane X receptor, cytochrome P450 3A4 enzyme, nuclear factor κB p65, nuclear factor κB p-p65, proliferating nuclear antigen, interleukin-6, interleukin-1β, tumor necrosis factor-α, inhibitor of nuclear factor κB α, cyclooxygenase 2, phosphorylated inhibitor of nuclear factor κBα, nuclear factor erythroid 2-related factor 2, kelch-like ECH-associated protein 1 antibody, phosphorylated nuclear transcription factor erythroid 2-related factor 2 were evaluated via western blot.
RESULTS AND CONCLUSION: Compared with the control group, cells in the sodium arsenite groups at various concentrations showed unclear cell membrane boundaries, reduced cytoplasm, decreased cell fusion rates, and widened intracellular space. Compared with the control group, the intracellular levels of reactive oxygen species and malondialdehyde were significantly increased in all sodium arsenite concentration groups (P < 0.05). The levels of interleukin-6, interleukin-1β, and tumor necrosis factor-α in the cell supernatant were also significantly increased (P < 0.05). The protein expressions of p-nuclear factor κB inhibitor protein α, nuclear factor κB p-p65, nuclear factor κB p65, tumor necrosis factor-α, and interleukin 1β were all increased (P < 0.05), and cell survival rates were all decreased (P < 0.05). The protein expressions of proliferation nuclear antigen, nuclear factor erythroid 2-related factor 2, phosphorylated nuclear factor erythroid 2-related factor 2 protein expression, and total superoxide dismutase activity were all decreased (P < 0.05). The mRNA and protein expressions of progesterone X receptor and cytochrome P450 3A4 enzyme were all decreased (P < 0.05). Compared with the control group, the intracellular glutathione reductase activity was reduced in the 20 and 30 μmol/L sodium arsenite groups (P < 0.05), and the expression of kelch-like ECH-associated protein 1, 
interleukin-6, and cyclooxygenase-2 proteins was increased (P < 0.05). To conclude, sodium arsenite may induce oxidative stress and inflammatory damage in liver cells by down-regulating the expression of pregnane X receptor, inhibiting the Nrf2 antioxidant pathway and activating the nuclear factor-κB inflammatory pathway, while also suppressing the expression of cytochrome P450 3A4 enzyme.

Key words:  sodium arsenite, liver injury, pregnane X receptor, oxidative stress, nuclear factor-κB signaling pathway, inflammatory response, CYP3A4, tissue construction

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