Countering effects of a combination of podophyllotoxin, podophyllotoxin β-D-glucoside and rutin hydrate in minimizing radiation induced chromosomal damage, ROS and apoptosis in human blood lymphocytes. (May 2016)

• Record Type: Journal Article
• Authors: Dutta, Sangeeta; Yashavarddhan, M.H.; Srivastava, Nitya Nand; Ranjan, Rajiv; Bajaj, Sania; Kalita, Bhargab; Singh, Abhinav; Flora, Swaran J.S.; Gupta, Manju Lata
• Is Part Of: Food and chemical toxicology. Volume 91(2016)
• Page Start: 141
• Page End: 150
• Publication Date: 2016-05
• Subjects: Gamma radiation -- Premature chromosome condensation -- Apoptosis -- Spindle microtubules -- Podophyllum hexandrum; DPBS Dulbecco Phosphate Buffer saline -- PBST Phosphate Buffer saline with Tween 20 -- DMF Dose Modifying Factor -- DCFDA 2′, 7′-Dichlorofluorescein diacetate -- DMF Dose Modifying Factor -- DMSO dimethylsulfoxide -- DNA Deoxyribonucleic Acid -- DSB Double strand break -- FITC Fluorescein isothiocyanate -- HPLC High pressure liquid chromatography -- LET Linear Energy Transfer -- MFI Mean Fluorescence Intensity -- MPF Maturation Promoting Factor -- PBMCs Peripheral blood mono nuclear cells -- PCC Premature Chromosome Condensation -- PEG Polyethylene glycol -- PHA-P Phytohemagglutinin-P -- PI Propidium Iodide -- PP Protein Phosphatase -- ROS Reactive Oxygen Species -- RPMI-1640 Roswell Park Memorial Institute-1640 medium -- TdT Terminal deoxynucleotidyl Transferase -- TUNEL TdT-mediated dUTP nick end labeling -- γ -H2AX Phosphorylated H2AX; Toxicology -- Periodicals; Food poisoning -- Periodicals; Food Poisoning -- Periodicals; Toxicology -- Periodicals; Toxicologie -- Périodiques; Intoxications alimentaires -- Périodiques; Food poisoning; Toxicology; Periodicals; Electronic journals; 615.9
• DOI: 10.1016/j.fct.2016.03.007
• Languages: English
• ISSNs: 0278-6915

Abstract

Abstract: The present study was conceptualized with the aim of developing a safe radioprotector for human application against radiation induced toxicity. For this study, a formulation (G-002M) prepared by combining three active principles isolated from rhizomes of Podophyllum hexandrum, was evaluated for its potential to protect genomic DNA of human blood cells exposed to different doses of radiation (5, 7&10Gy). Blood samples were pretreated (-1hr to exposure) with G-002M. Parameters of Premature Chromosome Condensation (PCC) assay like PCC-index, PCC-rings and PCC-fragments were used to estimate radiation induced chromosomal aberrations. Radiation (7Gy) induce ROS generation and its modulation by G-002M was determined by flow-cytometry and fluorescent microscopy while apoptosis (0, 2, 24&48 hr) was analyzed using TUNEL assay. Effect on spindle organization in G2/M arrested cells by all the three compounds individually was studied using immunofluorescence microscopy. Irradiation caused dose dependent linear increase in PCC-rings and fragments, while decline in PCC index. G-002M pretreatment significantly decreased these chromosomal aberrations at all the radiation doses and assisted cell survival as indicated by increased PCC index compared with radiation only group. Significant decrease in radiation induced intracellular ROS (45 ± 3%) and apoptosis (49.9%) was also exhibited by the formulation. On podophyllotoxin treatment, most of the cells have shown blocked spindles however, depicted normal arrangement. G-002M also demonstrated a highly significant Dose Modifying Factor or DMF (PCC-rings: 2.27 and PCC-fragments: 1.60). Present study based on many parameters along with DMF study, strongly suggests that G-002M is an effective formulation with a potential to minimize chromosomal damage even at very high radiation doses. Highlights: G-002M was found highly effective in protecting human lymphocytic DNA exposed to high doses of radiation. Radioprotective efficacy of G-002M can be attributed to ROS scavenging, reversible G2/M arrest and reduction in apoptosis. G-002M demonstrates a high DMF of 2.27 and 1.60 using cytogenetic parameters.