| 原雅艺,白佳玫,田连琛,等.硝酸铀酰对线粒体结构与功能的影响研究[J].中华放射医学与防护杂志,2025,45(12):1237-1244.Yuan Yayi,Bai Jiamei,Tian Lianchen,et al.Effects of uranyl nitrate on mitochondrial structure and function[J].Chin J Radiol Med Prot,2025,45(12):1237-1244 |
| 硝酸铀酰对线粒体结构与功能的影响研究 |
| Effects of uranyl nitrate on mitochondrial structure and function |
| 投稿时间：2024-12-13 |
| DOI：10.3760/cma.j.cn112271-20241213-00476 |
| 中文关键词: 铀 肾毒性 线粒体动力学 分子机制 |
| 英文关键词:Uranium Nephrotoxicity Mitochondrial dynamics Molecular mechanism |
| 基金项目:国家自然科学基金联合基金(U216720073) |
| 作者 | 单位 | E-mail | | 原雅艺 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006
华中科技大学生命科学与技术学院, 武汉 430071 | | | 白佳玫 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006 | | | 田连琛 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006 | | | 左雅慧 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006 | | | 王晓梅 | 苏州大学放射医学及交叉学科研究院, 苏州 215009 | | | 董娟聪 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006 | | | 程娇 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006 | | | 党旭红 | 中国辐射防护研究院放射医学与环境医学研究所, 太原 030006
放射医学协同创新中心, 苏州 215009 | dangxuhong005@163.com |
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| 中文摘要: |
| 目的 以人肾小管上皮细胞(HK-2)为研究对象，观察铀损伤对HK-2细胞线粒体形态结构的影响，研究铀损伤对HK-2细胞线粒体氧化应激以及能量代谢等功能的影响，从线粒体动力学角度探讨铀致HK-2细胞损伤的潜在机制。方法 给予HK-2细胞100 μmol/L硝酸铀酰溶液培养，分别作用不同时间(0、3、6、12、24 h)后，利用激光共聚焦显微镜观察硝酸铀酰染毒后细胞线粒体形态结构的变化；通过检测丙二醛(MDA)浓度、活性氧(ROS)、过氧化氢酶(CAT)活性、超氧化物歧化酶(SOD)活性，分析硝酸铀酰对HK-2细胞氧化应激水平的影响；通过检测线粒体膜电位(MMp)、线粒体呼吸链复合物Ⅳ、Ⅴ酶活力、三磷酸腺苷(ATP)含量，探讨硝酸铀酰对HK-2细胞能量代谢的影响。进一步利用Western blot方法分析硝酸铀酰染毒对线粒体动力学相关蛋白Mfn1、Mfn2、Opa1、Drp1、Fis1的表达的影响，筛选硝酸铀酰损伤敏感的蛋白。结果 100 μmol/L硝酸铀酰染毒3 h后，线粒体形态逐渐改变为明显的间断结构，6 h后ROS、MDA浓度显著升高(t=89.06、39.40，P＜0.05)，CAT、SOD酶活力显著降低(t=14.49、12.12，P＜0.05)，铀线粒体呼吸链复合物Ⅳ、Ⅴ酶活力、ATP含量、MMp均以时间依赖的方式降低(t=2.64~31.51，P＜0.05)，ATP关联呼吸能力下降。Drp1、Fis1表达水平呈现时间依赖性增加(tDrp1=14.02~185.70，P＜0.05；tFis1=19.72~165.10，P＜0.05)。结论 线粒体的结构损伤先于功能障碍，线粒体动力学异常可能是造成线粒体的结构损伤的原因之一。分裂蛋白Drp1和Fis1可能是铀损伤的关键靶点，为铀致肾毒性的机制研究提供了基础，为铀中毒的预防和治疗提供了新靶点。 |
| 英文摘要: |
| Objective To investigate the human renal tubular epithelial cells (HK-2 cells) to determine the effects of uranium exposure on the mitochondrial morphology, structure, and functions (e.g., oxidative stress and energy metabolism) in HK-2 cells and to explore the potential mechanisms underlying uranium-induced injury to HK-2 cells from the perspective of mitochondrial dynamics.Methods HK-2 cells were cultured using a 100 μmol/L uranyl nitrate solution for different times (0, 3, 6, 12, and 24 h). Then, changes in mitochondrial morphology and structure post-exposure were observed using a confocal laser scanning microscope. The effects of uranyl nitrate on oxidative stress levels in HK-2 cells were analyzed based on malondialdehyde (MDA) concentration, and reactive oxygen species (ROS) levels, as well as the activity of catalase (CAT) and superoxide dismutase (SOD). Meanwhile, the effects of uranyl nitrate on energy metabolism in HK-2 cells were investigated based on mitochondrial membrane potential (MMp), the enzyme activity of respiratory chain complexes Ⅳ and Ⅴ in mitochondria, and the adenosine triphosphate (ATP) content. Using Western blot (WB), the effects of uranyl nitrate exposure on the expression of mitochondrial dynamics-related proteins (Mfn1, Mfn2, Opa1, Drp1, and Fis1) were analyzed and the proteins sensitive to uranyl nitrate-induced damage were determined.Results At 3 h post-exposure, mitochondrial morphology in HK-2 cells gradually transitioned to a distinct fragmented structure. At 6 h post-exposure, the HK-2 cells exhibited significant increases in ROS and MDA concentrations (t = 89.06, 39.40, P < 0.05), distinct decreases in CAT and SOD enzyme activities (t = 14.49, 12.12, P < 0.05), and time-dependent reductions in the enzyme activities of respiratory chain complexes Ⅳ and Ⅴ in mitochondria, ATP content, and MMp (t = 2.64-31.51, P < 0.05), a decline in ATP-linked respiratory capacity, and time-dependent significant increased in the expression levels of proteins Drp1 and Fis1 (tDrp1 = 14.02-185.70, P < 0.05; tFis1 = 19.72-165.10, P < 0.05).Conclusions Mitochondrial structural damage precedes mitochondrial functional impairments, and abnormal mitochondrial dynamics might be a potential cause of the structural damage. Fission proteins Drp1 and Fis1 represent potential key targets of uranium-induced damage. These findings provide a basis for investigating the mechanisms behind uranium-induced nephrotoxicity and offer new targets for the prevention and treatment of uranium poisoning. |
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