基于微核试验的甲基丙二酸体外和体内遗传毒性研究

doi:10.3969/j.issn.1004-616x.2025.05.003

Abstract

Abstract: OBJECTIVE：This study was aimed to explore the genotoxicity of methylmalonic acid (MMA) based on in vitro and in vivo micronuclei assays， and potential factors influencing its genotoxicity. METHODS：For in vitro studies，human umbilical venous endothelial cells (HUVEC) were treated with 0， 0.1，0.5，1，5，and 10 mmol/L MMA at different times (7 d or 72 h)，and frequencies of micronuclei (MN)， nucleoplasmic bridge (NPB) and nuclear bud (NB) and genomic instability (GIN) were assessed using by the cytokinesis-blocked micronucleus assay (CBMN). In addition， the CBMN assay was employed to detect the impact of mitomycin C，homocysteine (Hcy) and folic acid on the genotoxicity of MMA in HUVEC cells. For in vivo studies，the genotoxicity of MMA was verified by the bone marrow micronucleus assay in CF-1 mice. RESULTS： Compared with control and groups of low-concentration MMA (0.1-5 mmol/L)，10 mmol/L MMA significantly increased rate of MN，NPB，and GIN in HUVEC (all P <0.01)，and the genotoxicity was derived from the intrinsic toxicity and acidic nature of MMA. When HUVEC were treated with 10 mmol/L MMA and 0.1 μg/mL mitomycin C together，rate of MN，NB and GIN were significantly elevated as compared to these of separately treated groups (P <0.05 and P <0.01). Furthermore，10 mmol/L MMA significantly reduced rate of MN and GIN induced by 100 μmol/L Hcy (P <0.05 and P <0.01). Compared with control，short-term (72 h) folic acid deficiency had no significant effect on genotoxicity of MMA， but supplementation of folic acid (45.4-90.7 μmol/L) significantly reduced rate of MN，NPB and GIN induced by 10 mmol/L MMA (P <0.05 and P <0.01). In CF-1 mice， high doses of MMA (200-400 mg/kg) exhibited significant acid-independent genotoxicity (MN rate) when compared with control (P <0.01). CONCLUSION：MMA showed intrinsic and acidmediated genotoxicity. Synergistic genotoxicity of MMA and mitomycin C was detected. Genotoxicity of MMA was mitigated by folic acid， and MMA ameliorated the genotoxicity of Hcy. This study provides new information for better understanding the genotoxicity of MMA and its intervention.

Key words: methylmalonic acid, genotoxicity, micronucleus test, CF-1 mice, HUVEC cells

CLC Number:

R394.6

LI Yuxiang, ZHU Yinlan, JI Qiuyue, LI Xiang, WANG Xu, ZHOU Tao, GUO Xihan. Genotoxicity of methylmalonic acid based on in vitro and in vivo micronucleus assays[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(5): 361-368.

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Genotoxicity of methylmalonic acid based on in vitro and in vivo micronucleus assays

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