草甘膦及其代谢物氨甲基膦酸对人外周血单个核细胞的影响  

Effect of glyphosate and its metabolite aminomethylphosphonic acid on human peripheral blood mononuclear cells

作　　者：张伟东[1] 周静芳 安迪 张伟威 阎彦[1] 张玲 党晓健 王娜[1] ZHANG Weidong;ZHOU Jingfang;AN Di;ZHANG Weiwei;YAN Yan;ZHANG Ling;DANG Xiaojian;WANG Na(Department of Pharmacy,the First Hospital of Qinhuangdao,Qinhuangdao,Hebei 066000,China)

机构地区：[1]秦皇岛市第一医院药学部,河北秦皇岛066000

出　　处：《中国工业医学杂志》2023年第3期200-205,共6页Chinese Journal of Industrial Medicine

基　　金：河北省医学科学研究课题(编号:20210364);河北省博士后基金资助项目(编号:B2021003046)。

摘　　要：目的 探讨不同浓度的草甘膦(glyphosate, GLY)及其代谢物氨甲基膦酸(aminomethylphosphonic acid, AMPA)对人外周血单个核细胞(peripheral blood mononuclear cells, PBMC)的影响。方法 将高(5 mmol/L)、中(0.5 mmol/L)、低(0.05 mmol/L)三种浓度的GLY及AMPA与人PBMC共孵育,采用逆转录-聚合酶链反应(real-time PCR,RT-PCR)和荧光探针法检测炎性因子白细胞介素(interleukin, IL)IL-1β和IL-6、肿瘤坏死因子-α(tumor necrosis factor, TNF-α)、细胞凋亡相关标志胱天蛋白酶(cell apoptosis related marker cystatin, Caspase)Caspase-3和Caspase-9、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2同源相关蛋白x(Bcl-2-associated x, Bax)、DNA甲基化标志DNA甲基转移酶(DNA methyltransferases, DNMT)DNMT1和DNMT3A、DNA去甲基化酶TET3(tet methylcytosine dioxygenase 3)、组蛋白甲基化修饰标志酶(euchromatic histone lysine methyltransferase, EHMT)EHMT1和EHMT2、组蛋白脱乙酰基酶(histone deacetylase, HDAC)HDAC3和HDAC5的基因表达,以及氧化应激标志活性氧(reactive oxygen species, ROS)和一氧化氮(nitric oxide, NO)的水平,观察GLY及AMPA对人PBMC的影响。结果 GLY及其代谢物AMPA均可诱导IL-6、IL-1β和TNF-α的分泌,其能力与浓度呈正相关,随着GLY、AMPA浓度的增加,Bcl-2基因表达降低,Bax、Caspase-3和Caspase-9基因表达增加。高浓度(5 mmol/L)GLY和AMPA可诱导DNMT1、DNMT3A和HDAC3的显著表达,及ROS和NO的含量增加,但不影响TET3、EHMT1、EHMT2基因的表达。结论 GLY及其代谢物AMPA对人PBMC具有潜在毒性,其效应可能与其参与炎性因子、细胞凋亡、氧化应激、DNA甲基化及组蛋白翻译后的修饰调控相关,且具有明显剂量依赖性。Objective To investigate the effect of glyphosate(GLY)and its metabolite aminomethylphosphonic acid(AMPA)on human peripheral blood mononuclear cells(PBMC).Methods Different concentrations(5、0.5、0.05 mmol/L)of GLY and its metabolite AMPA were co-incubated with human PBMC,and RT-PCR and fluorescent probe methods were used to detect the inflammatory factors(IL-1β,IL-6 and TNF-α),apoptosis-related markers(Caspase-3,Caspase-9,Bcl-2 and Bax),DNA methylation markers(DNMT1,DNMT3A and TET3),histone methylation-modified genes(EHMT1,EHMT2,HDAC3 and HDAC5),and the levels of oxidative stress markers(ROS,NO),to observe the effects of GLY and its metabolite AMPA on PBMC.Results The results showed that high concentration(5 mmol/L)of GLY and its metabolite AMPA can induce abnormal secretion of inflammatory factors(IL-1β,IL-6 and TNF-α)which was positively correlated with the concentration;with the increase of GLY or AMPA concentrations,the expression of Bcl-2 gene decreased,while the expressions of Bax,Caspase-3 and Caspase-9 genes increased.Additionally,high concentration(5 mmol/L)of GLY or AMPA can also induce significant rise of ROS and NO levels and significant genes expressions of DNMT1,DNMT3A and HDAC3,but did not affect the expressions of TET3、EHMT1 and EHMT2 genes.Conclusion The results suggested that GLY and its metabolite AMPA have some potential toxicity to PBMC,which may be related to its involvement in the regulation of inflammatory factors,apoptosis,oxidative stress,DNA methylation and histone post-translational modification,and has significant dose dependence.

关 键 词：草甘膦(GLY) 氨甲基膦酸(AMPA) 炎性因子 细胞凋亡 氧化应激 表观遗传学 

分 类 号：R994.3[医药卫生—毒理学]