中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (36): 9572-9579.doi: 10.12307/2026.920

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

诱导胰岛β细胞中铜死亡：靶向SLC39A14调控同型半胱氨酸的作用

李淑娟1，马凌桔2，3，李心如4，孙  睿4，马  奔4，祁  宁4，张启凡4，刘  莉4，柴月娥5，马胜超3，4   

  1. 宁夏医科大学总医院，1急诊科，2老年与特需医学科，宁夏回族自治区银川市  750004；3国家卫生健康委员会代谢性心血管疾病研究重点实验室，宁夏回族自治区银川市  750004；4宁夏医科大学检验学院，宁夏回族自治区银川市  750004；5贵州医科大学药学院，贵州省贵阳市  550000
  • 收稿日期:2025-11-06 修回日期:2026-03-25 出版日期:2026-12-28 发布日期:2026-05-25
  • 通讯作者: 马胜超，博士，副教授，国家卫生健康委员会代谢性心血管疾病研究重点实验室，宁夏回族自治区银川市 750004；宁夏医科大学检验学院，宁夏回族自治区银川市 750004 并列通讯作者：柴月娥，博士，副教授，贵州医科大学药学院，贵州省贵阳市 550000
  • 作者简介:李淑娟，女，1992年生，宁夏回族自治区银川市人，回族，2022年宁夏医科大学毕业，硕士，医师，主要从事心血管代谢方面研究。
  • 基金资助:
    国家自然科学基金青年项目(81900273)；项目负责人：马胜超；国家自然科学基金面上项目(82060139，82270492)，项目负责人：马胜超；宁夏自然科学基金优秀青年项目(2023AAC05035)，项目负责人：马胜超；宁夏医科大学校级重点项目(XZ2022004)；宁夏医科大学大学生创新创业训练计划项目(S202210752015)，项目负责人：田荣，指导老师：马胜超

Inducing cuproptosis in pancreatic β cells: regulating homocysteine via targeting SLC39A14

Li Shujuan1, Ma Lingju2, 3, Li Xinru4, Sun Rui4, Ma Ben4, Qi Ning4, Zhang Qifan4, Liu Li4, Chai Yuee5, Ma Shengchao3, 4   

  1. 1Emergency Department, 2Department of Geriatrics and Special Needs Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Key Laboratory of Metabolic Cardiovascular Disease Research of National Health and Wellness Committee, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 4Inspection College of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 5College of Pharmacy, Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • Received:2025-11-06 Revised:2026-03-25 Online:2026-12-28 Published:2026-05-25
  • Contact: Ma Shengchao, PhD, Associate professor, Key Laboratory of Metabolic Cardiovascular Disease Research of National Health and Wellness Committee, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Inspection College of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Co-corresponding author: Chai Yuee, PhD, Associate professor, College of Pharmacy, Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • About author:Li Shujuan, MS, Physician, Emergency Department of General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China (Young Scientist Program), No. 81900273 (to MSC); National Natural Science Foundation of China (General Program), Nos. 82060139 and 82270492 (both to MSC); Ningxia Natural Science Foundation (Excellent Young Program), No. 2023AAC05035 (to MSC); Key Project of Ningxia Medical University, No. XZ2022004; Ningxia Medical University College Student Innovation and Entrepreneurship Training Program, No. S202210752015 (to TR [principal investigator] and MSC [advisor])

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摘要：



文题释义：
PIWI相互作用RNA(piRNA)：是一类与PIWI家族蛋白特异性结合的小分子非编码RNA，主要在动物生殖细胞中发挥作用。piRNA与肿瘤等疾病的发生、发展也密切相关。在特定疾病状态下，piRNA会出现差异表达，具有作为新型生物标志物的潜力。
铜死亡：是指细胞在铜离子过量积累的条件下，依赖线粒体脂酰化蛋白的铜结合，引发一系列线粒体代谢紊乱与蛋白质聚集应激，最终导致细胞死亡的过程。其发生不依赖活性氧生成、铁代谢紊乱或凋亡通路，而是一种以线粒体依赖性蛋白质脂酰化修饰为核心的新型代谢相关死亡模式。

背景：高同型半胱氨酸血症与胰岛β细胞功能障碍以及抵抗有着密切关系，但其具体分子机制还有待研究。
目的：探讨piR-000699靶向调控SLC39A14影响同型半胱氨酸诱导胰岛β细胞铜死亡的潜在机制。
方法：体外培养小鼠胰岛β细胞株Min6，将其分为对照组(0 μmol/L 同型半胱氨酸)和同型半胱氨酸组(分别加入40，80，100 及120 µmol/L
同型半胱氨酸)，筛选出100 µmol/L的同型半胱氨酸为干预胰岛β细胞的最佳剂量。采用qRT-PCR检测同型半胱氨酸中piR-000699本底表达；Western blot检测同型半胱氨酸诱导胰岛β细胞中铜死亡相关蛋白FDX1、HSP70及piR-000699下游靶基因SLC39A14的表达；铜离子试剂盒测定胰岛β细胞内Cu2+水平。细胞转染分组：①对照组、同型半胱氨酸组(100 μmol/L)、同型半胱氨酸+mimics-NC组、同型半胱氨酸+piR-000699 mimics组；②对照组、同型半胱氨酸组(100 μmol/L 同型半胱氨酸)、同型半胱氨酸+inhibitor-NC组、同型半胱氨酸+piR-000699 inhibitor组。采用qRT-PCR检测转染piR-000699 inhibitor及piR-000699 mimics后细胞piR-000699的mRNA表达；Western blot检测转染 piR-000699 inhibitor及piR-000699 mimics后细胞FDX1、HSP70及SLC39A14蛋白的表达；铜离子试剂盒测定转染后细胞Cu2+表达水平。并构建SLC39A14的野生型和突变型双荧光素酶基因载体，验证piR-000699和SLC39A14之间的靶向结合关系。
结果与结论：①同型半胱氨酸诱导胰岛β细胞损伤的最佳浓度为100 μmol/L；②与对照组相比，同型半胱氨酸组中piR-000699 mRNA表达增高(P < 0.01)，铜死亡相关蛋白FDX1、HSP70及Cu2+表达均增高(P < 0.001)；③胰岛β细胞中干扰piR-000699后，同型半胱氨酸进一步下调FDX1、HSP70及SLC39A14蛋白水平(P < 0.01)，细胞中Cu2+表达水平降低(P < 0.01)；④胰岛β细胞过表达piR-000699后，同型半胱氨酸进一步上调了细胞中FDX1、HSP70及SLC39A14蛋白水平(P < 0.01)，细胞中Cu2+水平明显增高(P < 0.001)；⑤双荧光素酶结合报告实验提示：与mimics-NC比较，转染SLC39A14野生型质粒的胰岛β细胞中同时转染piR-000699 mimics后，其相对荧光素酶活性明显升高(P < 0.01)。结论：此次实验结果发现在同型半胱氨酸诱导胰岛细胞铜死亡过程中piR-000699可以通过靶向SLC39A14基因调控细胞铜死亡相关蛋白表达，研究为糖尿病的治疗和预防提供新的靶点和策略。
https://orcid.org/0009-0003-3993-3334(李淑娟)


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: piRNA, SLC39A14, 胰岛β细胞, 同型半胱氨酸, 铜死亡

Abstract: BACKGROUND: Hyperhomocysteinemia is closely associated with pancreatic β-cell dysfunction and insulin resistance, but its specific molecular mechanism remains to be elucidated.
OBJECTIVE: To investigate the potential mechanism by which piR-000699 targets SLC39A14 to regulate homocysteine-induced cuproptosis in pancreatic β cells.
METHODS: The mouse pancreatic β-cell line Min6 was cultured in vitro and divided into a control group (0 μmol/L homocysteine) and homocysteine groups (treated with 40, 80, 100, and 120 μmol/L homocysteine, respectively). A concentration of 100 μmol/L homocysteine was selected as the optimal dose for intervening in pancreatic β cells. qRT-PCR was used to detect the baseline expression of piR-000699 in homocysteine-treated cells. Western blot was used to detect the expression of cuproptosis-related proteins FDX1 and HSP70, as well as the piR-000699 downstream target gene SLC39A14, in homocysteine-induced pancreatic β cells. A copper ion assay kit was used to measure intracellular Cu²⁺ levels in pancreatic β cells. Cell transfection groups were as follows: (1) control group, homocysteine group (100 μmol/L), homocysteine+mimics-NC group, homocysteine+piR-000699 mimics group; (2) control group, homocysteine group (100 μmol/L homocysteine), homocysteine+inhibitor-NC group, homocysteine+piR-000699 inhibitor group. qRT-PCR was used to detect piR-000699 mRNA expression after transfection with piR-000699 inhibitor and piR-000699 mimics. Western blot was used to detect the protein expression of FDX1, HSP70, and SLC39A14 after transfection with piR-000699 inhibitor and piR-000699 mimics. A copper ion assay kit was used to measure Cu²⁺ expression levels after transfection. Wild-type and mutant dual-luciferase gene vectors for SLC39A14 were constructed to verify the targeted binding relationship between piR-000699 and SLC39A14.
RESULTS AND CONCLUSION: (1) The optimal concentration of homocysteine for inducing pancreatic β-cell injury was 100 μmol/L. (2) Compared with the control group, piR-000699 mRNA expression was increased in the homocysteine group (P < 0.01), and the expression of cuproptosis-related proteins FDX1 and HSP70, as well as Cu²⁺ levels, were all increased (P < 0.001). (3) After interfering with piR-000699 in pancreatic β cells, homocysteine further downregulated the protein expression of FDX1, HSP70, and SLC39A14 (P < 0.01), while intracellular Cu²⁺ expression levels were decreased (P < 0.01). (4) After overexpressing piR-000699 in pancreatic β cells, homocysteine further upregulated the protein expression of FDX1, HSP70, and SLC39A14 in the cells (P < 0.01), while intracellular Cu²⁺ levels were significantly increased (P < 0.001). (5) Dual-luciferase reporter assay results indicated that, compared with mimics-NC, the relative luciferase activity was significantly increased in pancreatic β cells co-transfected with the SLC39A14 wild-type plasmid and piR-000699 mimics (P < 0.01). In conclusion, these findings indicate that piR-000699 can modulate the expression of cuproptosis-related proteins by targeting the SLC39A14 gene during homocysteine-induced cuproptosis in pancreatic β cells, providing new targets and strategies for the treatment and prevention of diabetes.

Key words: piRNA, SLC39A14, pancreatic β cells, homocysteine, cuproptosis

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