二甲基甲酰胺对小鼠肝毒性及毒理学机制研究  

Hepatotoxicity and Toxicological Mechanism of N,N-dimethylformamide in Mice

作　　者：崔烨 王扬眉 桑灵丽 何逍 肖静[1] CUI Ye;WANG Yangmei;SANG Lingli;HE Xiao;XIAO Jing(Department of Occupational Medicine and Environmental Toxicology,School of Public Health,Nantong University,Nantong 226019,China)

机构地区：[1]南通大学公共卫生学院职业卫生与环境毒理学教研室,南通226019

出　　处：《生态毒理学报》2025年第2期225-232,共8页Asian Journal of Ecotoxicology

基　　金：南通市科技计划项目(MS2023002);国家自然科学基金资助项目(82173554)。

摘　　要：二甲基甲酰胺(N,N-dimethylformamide,DMF)是一种广泛应用的化工原料,具有明确的肝脏毒性,但损伤机制不明。本研究通过雄性小鼠亚慢性经口暴露观察DMF对肝脏的影响及潜在机制。雄性ICR小鼠(48只)经适应性喂养后随机分组,DMF经口暴露28 d,浓度分别为0、350、700及1400 mg·kg^(-1);记录体质量及脏器系数,苏木精-伊红(hematoxylin-eosin staining,HE)和原位末端标记术(TdT-mediated-dUTP nick end labeling,TUNEL)法观察小鼠肝脏病理学改变;显色法检测丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)及碱性磷酸酶(alkaline phosphatase,ALP)含量;实时荧光定量PCR(quantitative real-time PCR,qPCR)和蛋白免疫印迹法对Bcl-2同源结构域蛋白(bcl-2 homologous domain protein,Beclin1)、微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3B,LC3B)、螯合体1(sequestosome 1,SQSTM1/P62)、B细胞淋巴瘤/白血病-2(B cell lymphoma/leukemia-2,Bcl-2)以及Bcl-2相关X蛋白(BCL2-associated X,Bax)基因和蛋白表达水平进行定量。结果表明DMF造成小鼠肝质量增加(P<0.05)及形态改变;与0 mg·kg^(-1)组相比,暴露组ALT、AST和ALP随暴露浓度增加而增高,在1400 mg·kg^(-1)组达到最高,表现出剂量-效应关系(P<0.05);同样,随着DMF剂量增加,Bax和P62的mRNA水平显著升高,而Bcl-2、Beclin1和LC3B的mRNA水平显著降低(P<0.05),Western blot也检测到了以上蛋白的水平改变。以上结果提示DMF可能通过抑制细胞自噬引起肝细胞凋亡造成小鼠肝损伤。N,N-dimethylformamide(DMF),a widely utilized chemical precursor,exhibits well-characterized hepatotoxic properties.However,the precise mechanisms underlying its toxicity remain elusive.The present study aimed to explore the effects of DMF on hepatic function and to elucidate the potential mechanisms involved through subchronic oral exposure in male mice.Following an acclimatization period,48 male ICR mice were randomly allocated and subjected to oral administration of DMF at concentrations of 0 mg·kg^(-1),350 mg·kg^(-1),700 mg·kg^(-1),and 1400 mg·kg^(-1)for 28 days.Body weight and organ coefficients were monitored,and histopathological alterations in the liver were examined using hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL).Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and alkaline phosphatase(ALP)were quantified using colorimetric assays.Gene and protein expression l evels of Bcl-2 homology 3 domain-containing protein(Beclin1),microtubule-associated protein 1 light chain 3B(LC3B),sequestosome 1(SQSTM1/P62),B cell lymphoma/leukemia-2(Bcl-2),and BCL2-associated X(Bax)were determined using quantitative real-time PCR(qPCR)and Western blotting.The results demonstrated that DMF exposure led to an increase in liver weight(P<0.05)and morphological alterations in the mice.Compared to the control group,the exposed groups exhibited elevated levels of ALT,AST,and ALP,which correlated positively with increasing DMF concentrations,peaking at 1400 mg·kg^(-1),indicating a dose-dependent relationship(P<0.05).With increasing DMF doses,mRNA levels of Bax and P62 were significantly upregulated,whereas those of Bcl2,Beclin1,and LC3B were notably downregulated(P<0.05).Western blot analysis corroborated these findings at the protein level.These data suggest that DMF may induce hepatocellular apoptosis and subsequent hepatic injury in mice by suppressing cellular autophagy.

关 键 词：二甲基甲酰胺 肝损伤 细胞凋亡 自噬 

分 类 号：X171.5[环境科学与工程—环境科学]