棒曲霉素对人胃黏膜细胞体外毒性机理研究  

In vitro toxicity mechanism of patulin on human gastric epithelial cells

作　　者：徐慧 贾丽娜[1] 苏婧一 徐俐俐 左树娜 张伟[1,2] 杨倩[1] XU Hui;JIA Lina;SU Jingyi;XU Lili;ZUO Shuna;ZHANG Wei;YANG Qian(College of Science and Technology,Hebei Agricultural University,Cangzhou 061100,China;College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,China)

机构地区：[1]河北农业大学理工系,河北沧州061100 [2]河北农业大学食品科技学院,河北保定071000

出　　处：《食品与发酵工业》2020年第11期92-97,共6页Food and Fermentation Industries

基　　金：河北省自然科学基金项目(C2017204027);河北农业大学引进博士专项资助项目(ZD201623)。

摘　　要：研究了棒曲霉素(patulin,PAT)对人胃黏膜细胞(gastric epithelial cells,GES-1)的体外毒性作用,从细胞增殖、凋亡、氧化损伤和DNA损伤等方面对其毒性机理进行了初步探讨。采用四甲基偶氮唑盐法检测PAT对细胞增殖的毒性作用;通过荧光显微镜观察了细胞凋亡形态;利用反转录聚合酶链式扩增反应(reverse transcription-polymerase chain reaction,RT-PCR)实验考察了Bax和Bcl-2 mRNA相对表达量变化;检测了细胞内超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、谷胱甘肽(glutathione,GSH)和丙二醛(malonaldehyde,MDA)等氧化损伤指标变化;采用Western Blot技术对DNA双链断裂标志物γ-H2AX的表达量进行了测定。研究结果表明,当c(PAT)>1.0μmol/L时,PAT对GES-1细胞抑制率大于20%,荧光染色镜检可见明显的细胞凋亡现象;Bax mRNA转录水平表达量上调,而Bcl-2 mRNA转录水平表达量显著下调,表明GES-1细胞进入了凋亡阶段。同时,总SOD、CAT检测值降幅最高可达53.76%,GSH检测值降幅最高可达56.55%,而MDA最低增幅为139.59%;γ-H2AX表达量上调。研究结果表明PAT可显著抑制GES-1细胞增殖并诱导其凋亡,具有明显的量效关系。由研究结果推断PAT可能通过体外诱发氧化损伤和DNA损伤途径引起GES-1细胞凋亡。The toxicity mechanism of patulin(PAT)on human gastric epithelial cells(GES-1)was studied in vitro from the aspects of cell proliferation,apoptosis,oxidative damage and DNA damage.When the concentration of PAT was above 1.0μmol/L,the inhibition rate of PAT on GES-1 cells was greater than 20%.Meanwhile,the obvious apoptosis phenomenon was observed under fluorescence microscope.The relative expression level of Bax mRNA showed a significant up-regulation,whereas the relative expression level of Bcl-2 mRNA was distinctly down-regulated.At the same time,the activity of SOD and CAT decreased at most by 53.76%and the content of GSH reduced at most by 56.55%.The content of MDA increased at least by 139.59%.The expression of γ-H2AX was elevated with the increasing concentration of PAT.The results showed that PAT could significantly inhibit the proliferation and induce the apoptosis of GES-1 cells,with significant dose-effect relationship.It is concluded that PAT may induce apoptosis of GES-1 cells through oxidative damage and DNA damage.

关 键 词：棒曲霉素 人胃黏膜细胞 氧化损伤 DNA损伤 凋亡 

分 类 号：R99[医药卫生—毒理学]