现代预防医学

基因	引物序列
ND1	ND1-F:5’- TCCTAATGCTTACCGAACGAAA-3’
	ND1-R:5’- ATGGTAGATGTGGCGGGTTT-3’
Telo	Telo-F:5’-ACACTAAGGTTTGGGTTTGGTTTTGGTTTTGGTTAGTGT-3’
	Telo-R:5’-TGTTAGGTATCCCTATCCCTATCCCTATCCCTATCCCTAACA-3’
ACTB	ACTB-F:5’- GTGCACCTGACTCCTGAGGAGA-3’
	ACTB-R:5’- CCTTGATACCAACCTGCCCAG-3’

基因	引物序列
ND1	ND1-F:5’- TCCTAATGCTTACCGAACGAAA-3’
	ND1-R:5’- ATGGTAGATGTGGCGGGTTT-3’
Telo	Telo-F:5’-ACACTAAGGTTTGGGTTTGGTTTTGGTTTTGGTTAGTGT-3’
	Telo-R:5’-TGTTAGGTATCCCTATCCCTATCCCTATCCCTATCCCTAACA-3’
ACTB	ACTB-F:5’- GTGCACCTGACTCCTGAGGAGA-3’
	ACTB-R:5’- CCTTGATACCAACCTGCCCAG-3’

项目	对照组（n=18）	暴露组（n=78）	χ²（t）值	P值
吸烟人数			0.184^b	0.667
吸烟	9（50.0）	46（59.0）
不吸烟	9（50.0）	32（41.0）
年龄（岁）	35.212±3.238	36.236±6.814	0.715^a	0.476
饮酒人数			0.068^b	0.794
饮酒	8（44.4）	40（51.2）
不饮酒	10（55.6）	38（48.8）
端粒长度	1.087(1.008,1.178)	1.097(1.026,1.299)	601	0.348
mtDNA拷贝数	1.018(1.003, 1.041)	1.004(1.001, 1.020)	504	0.063
iAs(mg/g)	0.295 (0.073, 0.412)	1.998 (1.681, 2.400)	2	<0.001
MMA(mg/g)	0.066±0.387	2.112±0.517	15.788^a	<0.001
DMA (mg/g)	1.034±0.391	2.636±0.525	12.163^a	<0.001

项目	对照组（n=18）	暴露组（n=78）	χ²（t）值	P值
吸烟人数			0.184^b	0.667
吸烟	9（50.0）	46（59.0）
不吸烟	9（50.0）	32（41.0）
年龄（岁）	35.212±3.238	36.236±6.814	0.715^a	0.476
饮酒人数			0.068^b	0.794
饮酒	8（44.4）	40（51.2）
不饮酒	10（55.6）	38（48.8）
端粒长度	1.087(1.008,1.178)	1.097(1.026,1.299)	601	0.348
mtDNA拷贝数	1.018(1.003, 1.041)	1.004(1.001, 1.020)	504	0.063
iAs(mg/g)	0.295 (0.073, 0.412)	1.998 (1.681, 2.400)	2	<0.001
MMA(mg/g)	0.066±0.387	2.112±0.517	15.788^a	<0.001
DMA (mg/g)	1.034±0.391	2.636±0.525	12.163^a	<0.001

砷暴露人群中砷代谢产物与线粒体DNA拷贝数及端粒长度的差异相关性分析

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任有 , 卢承熙 , 凡则嘉 , 付美玲 , 孙榕蓉 , 温译涵 , 温丽芝 , 李舒婷

现代预防医学 | 环境与职业卫生 2026,53(11): 1981-1986

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现代预防医学 |环境与职业卫生 2026 , 53 (11) : 1981 -1986

砷暴露人群中砷代谢产物与线粒体DNA拷贝数及端粒长度的差异相关性分析

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任有, 卢承熙, 凡则嘉, 付美玲, 孙榕蓉, 温译涵, 温丽芝, 李舒婷

作者信息

云南省公共卫生与生物安全重点实验室、昆明医科大学公共卫生学院，云南昆明 650500

通讯作者:

李舒婷，E-mail：694199460@qq.com

作者简介:

任有（2003—），男，本科在读，研究方向：职业卫生与职业医学

Differential associations of arsenic metabolites with mitochondrial DNA copy number and telomere length in arsenic-exposed populations

You REN, Cheng-xi LU, Ze-jia FAN, Mei-ling FU, Rong-rong SUN, Yi-han WEN, Li-zhi WEN, Shu-ting LI

Affiliations

Yunnan Provincial Key Laboratory of Public Health and Biosafety, School of Public Health, Kunming Medical University, Kunming, Yunnan 650500, China

出版时间: 2026-06-10 doi: 10.20043/j.cnki.MPM.202601122

文章导航

摘要

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目的

在砷暴露人群中，探究单甲基砷酸（monomethylarsonous acid，MMA）、二甲基砷酸（dimethylarsinous acid，DMA）与线粒体DNA拷贝数、端粒长度的关联。

方法

采用横断面研究设计，选取云南省南部山区某焦炉厂的78名工人及其同村村民为研究对象，进行问卷调查并采集生物样本。使用氢化物发生-火焰原子吸收光谱法测定尿液中无机砷、MMA和DMA浓度，使用实时荧光定量聚合酶链式反应（polymerase chain reaction，PCR）测定线粒体DNA拷贝数和端粒长度。采用多重线性回归，以线粒体DNA拷贝数、端粒长度为因变量，经对数转化后的iAs、MMA、DMA为自变量进行统计分析。

结果

经多重线性回归模型校正混杂因素后，发现MMA与线粒体DNA拷贝数呈现显著正相关（， P=0.017），而DMA与线粒体DNA拷贝数呈显著负相关（， P=0.007），为排除多重共线性对模型稳定性的影响，进一步分析砷代谢指标，一级甲基化指数（primary methylation index， PMI）、二级甲基化指数（secondary methylation index， SMI）与线粒体DNA拷贝数的关系。结果显示，PMI与线粒体DNA拷贝数呈显著负相关（， P=0.041），而SMI则未呈现显著关联。此外，MMA和DMA与端粒长度的关联均未达到统计学显著性（P>0.05）。

结论

在砷代谢过程中，PMI与线粒体DNA拷贝数呈负相关，这可能与砷暴露诱发的线粒体氧化应激损伤以及过高的砷甲基化代谢引起的甲基化资源分配不均，导致线粒体DNA合成不足等多种因素共同作用有关。

关键词

砷暴露 / 线粒体DNA / 端粒长度 / 横断面研究

Abstract

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Objective

To investigate the associations of monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) with mitochondrial DNA copy number (mtDNA-CN) and telomere length (TL) in an arsenic-exposed population.

Methods

A cross-sectional study was conducted involving 78 workers from a coke oven plant and their fellow villagers from the same village in Southern Yunnan Province,China. Questionnaire surveys and biological sample collection were performed. Concentrations of inorganic arsenic (iAs), MMA, and DMA in urine were determined using hydride generation-flame atomic absorption spectrometry. mtDNA-CN and TL were measured by quantitative real-time polymerase chain reaction (PCR). Multiple linear regression analysis was performed with mtDNA-CN and TL as dependent variables and log-transformed iAs, MMA, and DMA as independent variables.

Results

After adjusting for confounders in the multiple linear regression model, MMA showed a significant positive association with mtDNA-CN (, P=0.017), while DMA showed a significant negative association (, P=0.007). To address potential multicollinearity and assess model stability, the relationships of arsenic methylation indices, including the primary methylation index (PMI) and secondary methylation index (SMI), with mtDNA-CN were further analyzed. The results showed that PMI was significantly negatively associated with mtDNA-CN (, P=0.041), whereas SMI showed no significant association. Furthermore, no statistically significant associations were observed between MMA or DMA and TL(P>0.05).

Conclusion

A negative correlation was observed between PMI and mtDNA-CN during arsenic metabolism. This may be attributed to the combined effects of arsenic-induced mitochondrial oxidative stress damage and potential insufficiency in mtDNA synthesis resulting from the disproportionate allocation of methyl resources due to highly active arsenic methylation.

Key words

Arsenic exposure / Mitochondrial DNA / Telomere length / Cross-sectional study

引用本文

任有, 卢承熙, 凡则嘉, 付美玲, 孙榕蓉, 温译涵, 温丽芝, 李舒婷. 砷暴露人群中砷代谢产物与线粒体DNA拷贝数及端粒长度的差异相关性分析. 现代预防医学, 2026 , 53 (11) : 1981 -1986 . DOI: 10.20043/j.cnki.MPM.202601122

You REN, Cheng-xi LU, Ze-jia FAN, Mei-ling FU, Rong-rong SUN, Yi-han WEN, Li-zhi WEN, Shu-ting LI. Differential associations of arsenic metabolites with mitochondrial DNA copy number and telomere length in arsenic-exposed populations[J]. Modern Preventive Medicine, 2026 , 53 (11) : 1981 -1986 . DOI: 10.20043/j.cnki.MPM.202601122

正文

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砷（As）是一种常见的类金属元素^[1]，主要以无机物形式（如As³⁺、As⁵⁺）存在于环境中，其环境行为及健康效应一直是环境科学与毒理学领域的研究热点。近一个世纪以来，采矿冶炼、化石燃料燃烧以及砷基杀虫剂的施用等人类活动显著加剧了砷在环境中的累积^[2]。相关数据显示，砷暴露已成为一个全球性环境问题，全球范围内超过2.3亿人正暴露于砷污染的环境中^[3]。

环境中的砷酸盐（As⁵⁺）可通过饮水、食物及职业暴露等途径进入人体。在体内，砷酸盐首先经生物还原作用转化为毒性更强的亚砷酸盐（As³⁺）；随后在肝脏内通过甲基转移酶介导的生物甲基化反应，逐步转化为单甲基砷酸（monomethylarsonous acid，MMA）、二甲基砷酸（dimethylarsinous acid，DMA）等有机代谢产物，最终以DMA为主要形态通过尿液排出体外^[4]。FARIHA等人^[5]的研究表明，长期砷暴露会显著增加癌症、皮肤病变、心血管疾病等多种疾病的发病风险。

砷的致病机制较为复杂，氧化应激是被广泛证实的核心机制之一。砷暴露可靶向损害线粒体电子传递链，导致活性氧（reactive oxygen species）过量生成，进而引发线粒体DNA（mitochondrial DNA，mtDNA）损伤，并加速端粒的缩短与其功能损害^[1,6]。

线粒体作为细胞的“能量工厂”，通过氧化磷酸化途径合成三磷酸腺苷（adenosine triphosphate，ATP）。其功能障碍被认为是介导机体衰老及多种年龄相关性疾病发生发展的关键环节^[7-8]。当线粒体功能受损时，电子传递链的偶联效率降低，导致活性氧大量累积。过量的活性氧不仅会对线粒体自身的DNA、蛋白质及脂质造成氧化损伤，还会扩散至细胞质和细胞核，对端粒产生直接的氧化应激损伤^[9]。

端粒是真核生物线性染色体末端的DNA - 蛋白质复合结构，其核心功能在于保护染色体末端免受降解与异常重组，从而维持基因组的结构完整性^[10]。受DNA复制末端缺陷机制的限制，端粒长度会随细胞分裂次数增加而逐渐缩短。当端粒缩短至临界阈值时，细胞将启动不可逆的细胞周期停滞程序，即细胞衰老^[11]。细胞衰老不仅是机体衰老的关键标志，也是神经退行性疾病、心血管疾病、骨骼肌肉疾病等多种年龄相关性疾病发生发展的重要病理基础^[12-13]。

综上所述，端粒长度和线粒体DNA拷贝数作为评估细胞衰老及功能障碍的关键生物标志物，在维持细胞稳态和机体健康方面扮演着至关重要的角色。砷暴露通过氧化应激等核心机制，直接或间接地调控端粒长度并损害线粒体功能，从而加速细胞衰老进程，促进多种疾病的发作和进展。

然而，现有研究主要集中于总砷暴露的生物学效应，对于砷特定代谢产物的独立作用机制研究尚显不足；此外，砷代谢产物之间的高度共线性也增加了探究其独立效应的方法学难度^[14]。本研究计划采用横断面研究设计，运用多重线性回归模型，系统地探究砷暴露人群中MMA、DMA与端粒长度及mtDNA拷贝数之间的关联。

1　材料与方法

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1.1　研究人群与设计

本研究受试者来自云南省南部山区，具体招募方案如下：暴露组纳入78名在焦炉冶炼厂工作时长≥3个月的职业性砷暴露工人；对照组纳入18名农业工人，其居住地距该冶炼厂十余公里，且近3个月内无农药接触史^[15-16]。由昆明医科大学经专业培训的研究人员对所有受试者进行体格检查和问卷调查，收集的信息包括年龄、性别、职业、岗位、工龄、吸烟饮酒史、既往疾病史及用药史等。同时，采集每位受试者外周静脉血5 ml及尿液20 ml，所有生物样本均置于4 ℃环境冷藏保存，并尽快转运至实验室进行后续分析。本研究已通过昆明医科大学伦理委员会审查批准(KMMU2021MEC036)。在研究实施前，研究人员向受试者充分告知研究目的、流程及相关风险，所有受试者均自愿签署知情同意书后纳入研究。

1.2　尿液中砷及其代谢产物的测定

尿液中砷及其代谢产物的测定采用氢化物发生-火焰原子吸收光谱法，依据WS/T29—1996标准进行。检测设备为原子吸收分光光度计AA-6800（Shimadzu Co., Kyoto,Japan生产）。标准品选用含砷量为(480±100) mg/L的冷冻干燥尿液标准参考物质SRM 2670（由美国NIST提供，位于盖瑟斯堡，马里兰州）。样品前处理采用混合酸消化法，随后加入硼氢化钠生成砷化氢，并将其输送到火焰燃烧器上的石英管中，在193.7 nm波长下进行原子吸收光谱法测定，以确定无机砷、MMA和DMA的浓度，并计算一级甲基化指数（primary methylation index，PMI）和二级甲基化指数（secondary methylation index，SMI），计算公式如下：

其中，tAs为总砷，As为无机神。

1.3　mtDNA拷贝数与端粒长度的检测

使用天根生物技术（北京）有限公司（中国北京）提供的试剂盒，从获取的受试者外周血样品中提取DNA；随后，通过实时荧光定量聚合酶链式反应（quantitative real-time PCR，qPCR）对每个样品进行三重复的检测。线粒体ND1基因、端粒相关序列及内参基因ACTB扩增使用的引物由上海生工生物工程股份有限公司（Sangon Biotech）合成，具体引物序列见表1。检测仪器为Roche LightCycler C96（曼海姆，德国）。PCR反应条件如下：在95 ℃预热10 min，随后进行95 ℃10 s、60 ℃10 s的35个循环，最后在60 ℃保持5 min结束。通过ΔCt值来反映mtDNA拷贝数与端粒长度，计算公式为ΔCt=ΔCtND1-ΔCtβ-ACTB；ΔCt=ΔCtTelo-、ΔCtβ-ACTB。

1.4　统计分析

调查问卷的数据通过双录入方式录入数据库，采用R 4.5.0统计学软件进行数据分析和绘图，采用GraphPad Prism 9.5.0进行绘图。对于对照组与暴露组的计量资料，若服从正态分布则使用t检验分析，否则采用Wilcoxon秩和检验；计数资料则使用χ²检验。通过lm函数以mtDNA拷贝数或端粒长度为因变量，以经对数转换后的iAs、MMA、DMA为自变量进行多重线性回归分析。为避免砷代谢产物之间多重共线性对模型稳定性的影响，以PMI、SMI进一步分析其与mtDNA拷贝数或端粒长度的关系。检验水准α=0.05。

2　结果

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2.1　研究对象的基本特征和砷暴露特征

本研究共招募96名研究对象，其中18名纳入对照组，78名纳入暴露组。人口学数据及砷暴露特征见表2。两组在年龄、吸烟率、饮酒率、端粒长度和mtDNA拷贝数方面差异无统计学意义（P>0.05）。然而，暴露组尿液中的iAs、MMA和DMA浓度均高于对照组（P<0.05）。见图1。

2.2　多重线性回归

为了揭示砷代谢物（iAs、MMA、DMA）的独立效应,本研究利用lm函数对其进行了多重线性回归分析。初步的单变量分析表明，经log10转换后的iAs、MMA、DMA与mtDNA拷贝数和端粒长度之间均未呈现出显著关联。基于统计显著性和生物学意义的考量，研究最终将MMA和DMA纳入多元线性模型。最终结果显示：MMA与mtDNA拷贝数呈显著正相关（

，P=0.017），而DMA与mtDNA拷贝数则呈显著负相关（

，P=0.007）。鉴于模型中MMA与DMA的方差膨胀因子（variance inflation factor,VIF）高达14.689，为保证模型稳健性，本研究转而采用其衍生指标PMI与SMI进行后续分析（VIF=1.479）。结果显示，PMI与线粒体DNA拷贝数呈显著负相关（

, P=0.041），而SMI无显著关联。此外，端粒长度与MMA、DMA之间均未发现统计学显著差异。见图2。

3　讨论

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3.1　主要发现

本研究通过横断面设计，探讨了砷暴露人群中无机砷代谢产物（MMA、DMA）与mtDNA拷贝数及端粒长度的关联。主要发现暴露组尿中的砷、MMA和DMA浓度均显著高于对照组（P<0.05），但两组在年龄、吸烟率、饮酒率、mtDNA拷贝数和端粒长度方面差异无统计学意义（P>0.05）。采用多重线性回归校正混杂因素后，发现MMA与mtDNA拷贝数呈显著正相关（

, P=0.017），而DMA则呈显著负相关（

, P=0.007）。鉴于此模型中MMA与DMA的VIF高达14.689，存在严重共线性，为获取稳健估计，本研究转而采用其衍生指标PMI与SMI进行分析。结果显示，PMI与mtDNA拷贝数呈显著负相关（

, P=0.041），而与SMI无显著关联。此外，未发现MMA或DMA与端粒长度存在统计学关联。

3.2　砷代谢物与mtDNA拷贝数、端粒长度相关性机制探讨

暴露组尿中的砷、MMA和DMA浓度均显著高于对照组，这确证了研究分组的有效性，表明人群确实存在砷暴露及代谢差异，同时也证实了这些生物标志物对近期暴露的敏感性。然而，两组在mtDNA拷贝数和端粒长度的均值上并未表现出显著差异。根据后续研究结果可知，这可能是由于MMA和DMA对mtDNA拷贝数产生的正负效应相互叠加所致。换言之，尽管砷暴露尚未在群体水平上引发宏观、显著的DNA损伤，但其影响可能已通过更精细的、代谢产物特异性的方式显现出来。

mtDNA拷贝数作为评估线粒体功能和细胞能量代谢的重要标志物，其变化可能与氧化应激、DNA合成障碍等生物通路密切相关。一方面，研究表明，砷暴露通过诱发氧化应激导致线粒体损伤，而mtDNA因缺乏组蛋白保护且靠近活性氧生成位点，对氧化损伤尤为敏感^[17]。因此，PMI与mtDNA拷贝数的负相关很可能反映了砷暴露对线粒体基因组完整性的损害。另一方面，砷主要以As³⁺、As⁵⁺的形式被人体所摄取，在代谢过程中，As⁵⁺首先被谷胱甘肽和硫氧化还原蛋白还原为毒性更强的As³⁺。随后，在肝脏经过一系列甲基化反应，依次生成MMA和DMA，最终以DMA为主要代谢物随尿液排出体外^[18]。而PMI在砷代谢中具有重要的生物学意义，其主要反映了个体将无机砷甲基化为MMA的能力^[19]。而高水平的砷甲基化往往需要消耗更多的S-腺苷甲硫氨酸（S-adenosyl methionine，SAM）作为甲基供体^[20]。此外，SAM不仅是砷代谢过程中关键的甲基供体，其在机体DNA合成的过程中也发挥着至关重要的作用^[21]，综上所述，本研究推测PMI与mtDNA拷贝数的负相关，可能与砷暴露诱发的线粒体氧化应激损伤以及过高的砷甲基化代谢引起的甲基化资源分配不均，导致mtDNA合成不足等多种因素共同作用有关。

在本研究中，经多重线性回归后，端粒长度与iAS、MMA、DMA未显示出显著的统计学意义，这可能与砷对端粒长度影响的复杂性密切相关。BORGHINI等人^[22]的研究表明，砷可能通过引发氧化应激增加活性氧，从而导致端粒长度缩短。然而，在孟加拉国进行的一项大型研究却提示，砷具有延长端粒长度的作用，这可能与端粒酶的上调及新端粒DNA序列的合成有关^[23]。此外，ZHANG等人^[24]的一项涵盖1 469人的孟加拉队列研究同样得出砷暴露水平与端粒长度无显著相关性的结论。相较于mtDNA拷贝数，端粒长度是一个变化更为缓慢的指标。mtDNA拷贝数能够对急性氧化应激和能量需求做出相对快速的响应，而端粒缩短则通常是一个长期且累积的过程^[25]。本研究的横断面设计可能未能捕捉到砷暴露对端粒的细微或长期影响。

3.3　优势与局限性

本研究的优势在于：在砷暴露人群中，探讨了关键砷代谢产物（MMA与DMA）分别与mtDNA拷贝数及端粒长度的独立关联。这一研究视角从“总砷负荷”拓展至“代谢产物”，为深入理解砷的毒作用模式提供了新视角，并为揭示不同代谢路径在砷介导细胞损伤中的特异性作用机制提供了初步证据。此外，mtDNA拷贝数有望成为砷致早期细胞效应的高敏感性生物标志物，为后续深入研究奠定了坚实基础。

本研究的局限性如下：（1）本研究属于横断面研究，难以进行因果推断。因此，相关研究结果需在前瞻性队列研究中进一步验证，且相关机制仍需通过实验性研究加以探讨。（2）本研究的样本量相对较小，这可能在一定程度上限制了结果的统计效力和结论的普遍适用性。

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参考文献

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2026年第53卷第11期

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doi: 10.20043/j.cnki.MPM.202601122

接收时间：2026-01-10
首发时间：2026-11-02
出版时间：2026-06-10

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收稿日期：2026-01-10

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“兴滇英才支持计划”项目经费支持(YNQR-QNRC-2020-103)

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云南省公共卫生与生物安全重点实验室、昆明医科大学公共卫生学院，云南昆明 650500

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李舒婷，E-mail：694199460@qq.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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