Integrative Assessment of the Novel Radioprotective Efficacy of Cardamonin: A Comprehensive In Vitro and In Silico Analysis

Overview

Arkansas for Medical Sciences Current radiobiology research mainly focusses to identify efficient radioprotective agents that can protect normal tissues from radiation-induced injury due to accidental exposures. Recently, cardamonin has gained interest for its potential antioxidant, an ti-inflammatory, anti-cancer activity and biosafety properties The main purpose of this project is to decipher the novel radioprotective effects of cardamonin by molecular docking evaluations and in vitro experiments. Clone 9, HEK 293 and WI38 cells were exposed to 8 Gy gamma irradiation after treatment with or without Cardamonin (5 µM) and cell morphology and intracellular ROS generation (DCFDA) were assessed. Further studies were done in clone 9 cells. Gene (RTPCR) a nd protein expression (Western Blotting) of Nrf2, HO -1 SOD2, Catalase and GPx were evaluated. Apoptosis was measured by live cell imaging (Incucyte) and protein expression. Molecular docking studies were done with Discovery studio and autodock-vina computational docking software. Pretreatment of normal liver, kidney and lung cells with cardamonin significantly prevented cell death, and reduced intracellular ROS levels after irradiation. Cardamonin pretreated clone 9 cells significantly upregulated the transcription factor Nrf2 and maintained antioxidant status, which protected the cells against radiation insult. Radiation-induced apoptosis was significantly prevented by inhibiting the activation of caspases 3, 7 and 9. Docking studies revealed the potent anti-apototic, anti-inflammatory and anti-fibrotic efficacy of cardamonin. In conclusion, cardamonin activated the Nrf2 dependent antioxidant pathway and prevented apoptotic cell death after irradiation. The in vitro and in silico studies provided significant evidence that cardamonin may serve as a promising radiation countermeasure to mitigate radiation injury.