邻苯二甲酸二辛酯对酿酒酵母BY4742细胞的毒性作用  

Toxic effects of dioctyl phthalate on Saccharomyces cerevisiae BY4742 cells

作　　者：徐应丽 邹伟 XU Ying-li;ZOU Wei(School of Public Health,Kunming Medical University,Kunming,Yunnan 650500,China)

机构地区：[1]昆明医科大学公共卫生学院,云南昆明650500

出　　处：《环境与健康杂志》2024年第11期954-959,F0003,共7页Journal of Environment and Health

基　　金：云南省教育厅科学研究基金(2023Y0614);云南省科技厅科技计划(202301AY070001-194)。

摘　　要：目的探讨邻苯二甲酸二辛酯(DEHP)对酿酒酵母BY4742细胞的毒性作用,为DEHP的毒性作用机制的研究提供了基础。方法以0.0 mg/L(对照)、1.0 mg/L、10.0 mg/L、100.0 mg/L邻苯二甲酸二辛酯对酿酒酵母BY4742细胞进行染毒,观察细胞的生长情况;以0.00 mg/L(对照)、0.10 mg/L、0.25 mg/L、0.50 mg/L邻苯二甲酸二辛酯对酿酒酵母BY4742细胞染毒,对细胞的骨架、细胞核、细胞凋亡进行染色观察,采用qRT-PCR实验检测细胞骨架相关基因的表达变化。结果与对照组比较,除1.0 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的相对生长率略有升高(P>0.05)外;10.0 mg/L和100.0 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的相对生长率均较高,差异有统计学意义(P<0.05)。通过斑点实验得出,与对照组0.00 mg/L相比较,随着邻苯二甲酸二辛酯浓度增大,同一梯度下酵母菌落逐渐变稀疏,即生长受到抑制。表明不同浓度1.0 mg/L、10.0 mg/L、100.0 mg/L邻苯二甲酸二辛酯均会抑制酵母细胞生长;且随着邻苯二甲酸二辛酯浓度的升高,酵母菌落生长情况变得愈加稀疏,即抑制生长效果愈明显。当邻苯二甲酸二辛酯浓度达到100.0 mg/L时,菌落变得更加稀疏,仅有少量酵母BY4742细胞生长。与对照组相比较,经0.10 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的细胞核略微缩小,但无明显结构变化,细胞核异常率略有增加(P>0.05);0.25 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的细胞核缩小,细胞核异常率明显增加(P<0.05);0.50 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的细胞核会呈现散点状,结构不完整,细胞核异常率明显增高(P<0.05)。随着邻苯二甲酸二辛酯染毒浓度的升高,BY4742细胞的细胞核出现了核碎裂,形成点块状大小不一的核碎片,核异常率呈上升趋势。与对照组比较,0.10、0.25、0.50 mg/L邻苯二甲酸二辛酯染毒组BY4742细胞的早期、晚期凋亡细胞率明显升高,差异均有统计学�Objective To understand the toxicity and mechanism of dioctyl phthalate to saccharomyces cerevisiae BY4742 cells.was studied by using different concentrations of dioctyl phthalate on the growth,skeleton and apoptosis of yeast cells.Methods The cells of saccharomyces cerevisiae BY4742 were exposed to dioctyl phthalate at the doses of 0.00 mg/L,1.00 mg/L,10.0 mg/L and 100.0 mg/L respectively,and the growth of the cells was observed.The cells of saccharomyces cerevisiae BY4742 were exposed to dioctyl phthalate at the doses of 0.00 mg/L,0.10 mg/L,0.25 mg/L and 0.50 mg/L respectively,and the cytoskeleton,nucleus and apoptosis of the cells were observed by staining,and the expression of cytoskeletons-related genes was detected by qRT-PCR.Results Dioctyl phthalate had no obvious effect on the growth of saccharomyces cerevisiae BY4742 cells at a low concentration of 1.00 mg/L(P>0.05),that is,no obvious acute toxic effect,but had obvious inhibitory effect on the growth of saccharomyces cerevisiae BY4742 cells at a high concentration of 10.0 mg/L and 100.0 mg/L(P>0.05).After exposure to dioctyl phthalate at low concentrations of 0.10 mg/L,0.25 mg/L,and 0.50 mg/L,BY4742 saw nuclear fragmentation,the formation of nuclear fragments with different size patches,the cytoskeleton showed spots and plaques,and the number of apoptosis increased.The transcription levels of cytoskeleton-related proteins Arc15p,Las17p and Crn1p were significantly down-regulated by qRT-PCR compared with the control group(P<0.05).With the increase of dioctyl phthalate,the skeletal abnormality rate of BY4742 cells increased,and the relative transcription levels of protein genes Arc15,Las17 and Crn1 in the skeletal of the cells were significantly down-regulated.Conclusion Dioctyl phthalate has toxic effect on BY4742 cells,and the mechanism may be related to the destruction of the nuclear structure of cells and the alteration of gene transcription levels of cytoskeletal proteins.

关 键 词：酿酒酵母 基因 邻苯二甲酸二辛酯 细胞骨架 

分 类 号：R994.6[医药卫生—毒理学]