title: 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.
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.
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Dutta
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: M H
authors.initials: MH
authors.lastname: Yashavarddhan
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Nitya Nand
authors.initials: NN
authors.lastname: Srivastava
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Rajiv
authors.initials: R
authors.lastname: Ranjan
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Sania
authors.initials: S
authors.lastname: Bajaj
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Bhargab
authors.initials: B
authors.lastname: Kalita
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India.
authors.forename: Abhinav
authors.initials: A
authors.lastname: Singh
authors.affiliation: Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Jhansi Road, Gwalior 474002, India.
authors.forename: Swaran J S
authors.initials: SJ
authors.lastname: Flora
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences (INMAS), Defence Research and Development Organization, S. K. Mazumdar Marg, Delhi 110054, India. Electronic address: drmanjugupta2003@yahoo.com.
authors.forename: Manju Lata
authors.initials: ML
authors.lastname: Gupta
mesh_terms: D017209:Apoptosis; D002869:Chromosome Aberrations; D005960:Glucosides / Q000737:chemistry*; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D011034:Podophyllotoxin / Q000737:chemistry / Q000494:pharmacology*; D011829:Radiation Dosage; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012431:Rutin / Q000494:pharmacology*
chemical_list: D005960:Glucosides; D017382:Reactive Oxygen Species; D012431:Rutin; D011034:Podophyllotoxin
keywords: Apoptosis; Gamma radiation; Podophyllum hexandrum; Premature chromosome condensation; Spindle microtubules