Mukul Prasad1, Stephen Charles Bronson2, Tushar Warrier1, Agnihotram Badarinath1, Shivam Rai1, Kaushal Baid1, Sneha Sitaraman1, Alex George1, Anand Moses2, Radha Saraswathy1, Ranganathan Vasuki2, Alagianambi Shanmugam2
1Biomedical Genetics Research Lab, Division of Biomolecules and Genetics, School of Bio Sciences and Technology, VIT University, Vellore, India.
2Institute of Diabetology, Madras Medical College & Rajiv Gandhi Government General Hospital, Chennai, Tamil Nadu, India.
Background: The increasing incidence of Type 2 diabetes mellitus globally has collaterally increased the incidence of diabetes-associated complications such as neuropathy. Oxidative stress induced DNA damage is one of the mechanisms implicated in the pathogenesis of diabetic complications. Here we aimed to evaluate the extent of DNA damage in diabetes patients with and without clinical neuropathy using the Cytokinesis Block Micronucleus Cytome assay, in a group of South Indian population. Materials and Methods: The Cytokinesis Block Micronucleus Cytome assay was performed in lymphocyte cultures of 42 type 2 diabetes patients (22 with neuropathy and 20 without neuropathy) and 42 age and sex matched controls. Nuclear aberrations like Nuclear Buds, Nucleoplasmic Bridges and Micronuclei were analyzed. Results: The frequency of nuclear aberrations in diabetes patients with neuropathy was higher than compared to diabetes patients without neuropathy. The mean frequencies of nuclear aberrations per cell in diabetes patients with neuropathy and without neuropathy were 0.02 ± 0.02 and 0.01 ± 0.01, respectively. This was significantly higher than in the controls (0.002 ± 0.002) (P < 0.0001). An increasing trend of nuclear aberrations in correlation with the duration of diabetes was observed. Conclusion: This study highlights the use of the Cytokinesis Block Micronucleus Cytome assay as a potent tool for the identification of DNA damage, which may prove to be useful biomarker to assess the severity diabetes-associated complications such as neuropathy. Implementation of this technique at the clinical level would potentially enhance the quality of management of patients with diabetes and its complications like neuropathy.
Keywords: CBMN Cyt assay, DNA damage, diabetic neuropathy, nuclear aberrations, oxidative stress, Type 2 diabetes mellitus.