Abstract

RUTIN THE BIOFLAVONOID FROM EUPHORBIA HIRTA AMELIORATES GAMMA RADIATION INDUCED RADIOTOXICITY IN HUMAN LYMPHOCYTES IN VITRO

Shabana S.* and Ramesh C.

ABSTRACT

The current study was executed to isolate and evaluate flavonoid from Euphorbia hirta for in vitro radioprotective potentials against gamma radiation induced radiotoxicity in human lymphocytes. The authenticated Euphorbia hirta plant material was defatted and subjected to methanol extraction using soxhlet apparatus. The prepared methanol extract of Euphorbia hirta was processed in column chromatography to isolate bioflavonoids. Isolated flavonoid rutin was tested for in vitro radioprotective potentials by micronucleus test, DNA fragmentation assay, superoxide free radical scavenging method and apoptic assay in human lymphocytes exposed to gamma radiation. Pretreatment of lymphocytes with Rutin (50μg/ml) exposed to 2 Gy gamma radiation caused significant reduction of micronuclei frequency when compared to the normal control set of lymphocytes assessed by cytokinesis blocked micronucleus assay. Similarly, treatment of human lymphocytes with rutin prior to gamma radiation exposure exhibited significantly reduced DNA damage as tested by comet parameters which includes Olive tail moment and percent tail DNA. Further, addition of rutin (50μg/ml) to lymphocytes before gamma irradiation considerably declined the percentage of apoptotic bodies investigated microscopic method and by DNA ladder assay. Similarly, there was significant inhibition of reactive oxygen species concentrations induced by gamma radiation by rutin. The results of our study demonstrated the protective effect of rutin against gamma radiation produced DNA damage and anti-apoptotic activity in lymphocytes isolated from human blood, which may be partially contributed to inhibition of free radicals produced by gamma radiation and also by the attenuation of gamma radiation induced oxidative stress.

Keywords: Radioprotective, Euphorbia hirta, Rutin, Lymphocytes, Gamma radiation, Radiotoxicity, Micronuclei, Apoptosis.