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评价三种食品添加剂对人肝细胞系的细胞毒性和诱导凋亡作用。

Evaluation of the Cytotoxicity and Apoptotic Induction in Human Liver Cell Lines Exposed to Three Food Additives.

机构信息

Faculty of Biotechnology, October University for Modern Sciences and Arts, (MSA), Wahat Road, 6th of October City, Egypt.

Department of Genetics, Faculty of Agriculture, Ain Shams University, Shubra el-Kheima, Egypt.

出版信息

Recent Pat Food Nutr Agric. 2020;11(3):193-201. doi: 10.2174/2212798411666200217124630.

Abstract

BACKGROUND

Rapid lifestyle, especially among people living in urban areas, has led to increasing reliance on the processed food market. Unfortunately, harmful effects caused by the excessive use of food additives in such type of industry are often neglected.

OBJECTIVE

This proposal investigates in vitro cytotoxic and apoptotic effects of three food preservatives commonly consumed in daily meals; sodium sulphite, boric acid, and benzoic acid.

METHODS

The effect of the three preservatives on cell viability was tested on two different cell lines; normal liver cell line THLE2 and human hepatocellular carcinoma cancer cell line HepG2 using MTT assay. Cell cycle arrest was measured using flow cytometry by propidium iodide. Measurement of expression levels of two central genes, p53 and bcl-2 that play key roles in cell cycle and apoptosis was carried out in HepG2 cells using real time-PCR.

RESULTS

Although the effect was more significantly realized in the HepG2 cell line, the viability of both cell lines was decreased by all of the three tested compounds. Flow cytometric analysis of HepG2 cells treated with sodium sulphite, boric acid, and benzoic acid has revealed an increase in G2/M phase cell cycle arrest. In Sodium sulphite and boric acid-treated cells, expression levels of p53 were up-regulated, while that of the Bcl2 was significantly down-regulated. On the other hand, Benzoic acid has shown an anti-apoptotic feature based on the increased expression levels of Bcl-2 in treated cells.

CONCLUSION

In conclusion, all of the tested compounds have decreased the cell line viability and induced both cell cycle arrest and apoptotic events indicating their high potential of being cytotoxic and genotoxic materials.

摘要

背景

快速的生活方式,尤其是在城市居民中,导致人们越来越依赖加工食品市场。不幸的是,这种行业中过度使用食品添加剂所带来的有害影响往往被忽视。

目的

本研究旨在调查三种日常生活中常用的食品防腐剂——亚硫酸钠、硼酸和苯甲酸——对体外细胞的细胞毒性和凋亡作用。

方法

采用 MTT 法检测三种防腐剂对两种不同细胞系(正常肝细胞系 THLE2 和人肝癌细胞系 HepG2)的细胞活力的影响。采用碘化丙啶(PI)流式细胞术检测细胞周期阻滞。采用实时 PCR 法检测 HepG2 细胞中两个在细胞周期和凋亡中起关键作用的核心基因 p53 和 bcl-2 的表达水平。

结果

虽然在 HepG2 细胞系中效果更为显著,但三种测试化合物均降低了两种细胞系的活力。用亚硫酸钠、硼酸和苯甲酸处理 HepG2 细胞的流式细胞术分析显示 G2/M 期细胞周期阻滞增加。在亚硫酸钠和硼酸处理的细胞中,p53 的表达水平上调,而 Bcl2 的表达水平显著下调。另一方面,苯甲酸表现出抗凋亡特征,因为处理细胞中 Bcl-2 的表达水平增加。

结论

综上所述,所有测试化合物均降低了细胞系活力,并诱导细胞周期阻滞和凋亡事件,表明它们具有很高的细胞毒性和遗传毒性物质潜力。

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