June 2016 Vol. 4 No.6

Search Pubmed for articles by:
 

Original Research Article

The use of GST-µ Gene and Isoenzymes as Biomarkers to Evaluate the Mutagenicity and Hepatic Carcinogenicity in the Mouse by Carmoisine ‘E122’

Mohamed S. Salama¹, Mohamed A. Ismail^2*, Mohamed A. Shahin² and Heba M. Yassin²

¹Faculty of Science, Ain Shams University, Cairo, Egypt
²Department of Biology, Faculty of Education, Ain Shams University, Cairo, Egypt

*Corresponding Author E-mail: mohesmael@hotmail.com

Accepted June 13, 2016

Abstract

In the past few years there has been a sharp rise in the use of food and beverages colourants to improve their characteristics. More worryingly is the commonly used colourant in most pharmaceutical syrups and coating of pills and capsules, particularly in children medicines. So, the present work was planned to study the effects of carmoisine as a common synthetic food-drug colorant in Egypt on the liver of the male mouse. The work plan was designed to cover four parameters: Morphological or external symptoms, protein expression and seven isozymes changes that might appear on the carmoisine-treated animals. Besides, molecular studies involving two main aspects: firstly, application of restriction endonuclease (RE) digestion (SmaΙ, XbaΙ, HindIII) on genomic DNA of mice liver in both treated and control groups. Secondly, PCR analysis to investigate the capability of carmoisine to induce mutations in GST-µ gene in treated groups comparable to control ones. According to FAO/WHO, the mice were given the Acceptable Daily Intake (ADI) dose of carmoisine (0.02mg/kg b.wt). This dose was also given in half and double fashions for 45 and 90 days for each. The results revealed that the carmoisine caused many morphological symptoms reflected mainly by agitation and a marked loss of their body weights in a significant decrease (p<0.05). The protein banding patterns in the liver carmoisine-treated mice exhibited distinct collapses when analyzed by native-PAGE and SDS-PAGE. The study also detected several changes in the profiles of the seven isozymes analysis for all treated groups. The RE detected liver DNA alterations. However, the GST-µ gene was not affected by this treatment, whereas the band at ~160 bp was found in entirely treated and control groups. Such phenomenon reflected the absence influences of carmoisine as a carcinogenic agent as dosages and periods used in this study on male mice, so further sequencing investigation is required for the evaluation of the safety of that E122.

Keywords: Carmoisine, GST-µ, Isozymes, Liver, Mice, PCR, Protein Expression, Restriction Endonucleases.