Ferulic Acid Reduces Cell Viability through Its Apoptotic Efficacy: An In vitro Approach
Murugaraj Manoj Prabhakar
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar – 608 002, Tamilnadu, India.
Shanmugam Manoharan *
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar – 608 002, Tamilnadu, India.
Tharmaraj Rejitharaji
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar – 608 002, Tamilnadu, India.
Ranganathan Selvasundaram
Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar – 608 002, Tamilnadu, India.
Villianur Ibrahim Hairul Islam
Pondicherry Centre for Biological Sciences, Boomyinpet, Puducherry-5, India.
*Author to whom correspondence should be addressed.
Abstract
Aim: Ferulic acid, a well known dietary phenolic antioxidant, possesses diverse pharmacological and biochemical effects, including anti-inflammatory, hepatoprotective, antidiabetic and anticancer properties. The present study explores the cytotoxic potential of ferulic acid using Hep-2 cell line by analyzing its effect on cell viability, reactive oxygen species generation, apoptotic induction, nuclear damage, DNA fragmentation and expression of apoptosis related proteins.
Materials and Methods: The effect of ferulic acid (2.5, 5, 10, 20 and 40 µg/ml) on Hep-2 cells viability for 24 hr was determined by MTT assay. To substantiate the cytotoxic effect of ferulic acid, the intracellular ROS level was determined using DCFH-DA assay; apoptosis by dual staining; nuclear damage by DAPI staining; DNA fragmentation by using agarose gel electrophoresis; apoptosis related proteins by western blotting.
Results: Ferulic acid significantly inhibited the Hep-2 cell growth in a dose dependent manner and ferulic acid treated Hep-2 cells exhibited features of apoptosis and increase in nuclear damage and DNA fragmentation. We also observed excess reactive oxygen species generation in ferulic acid treated Hep-2 cells. Apoptosis related proteins (p53, Bcl-2, Bax, Caspase 3 & Caspase 9) were significantly modulated in favour of programmed cell death in ferulic acid treated cells.
Conclusion: We thus conclude that the cytotoxic potential of ferulic acid might be due to its role in apoptosis induction, excessive ROS generation and DNA fragmentation in Hep-2 cells.
Keywords: Apoptosis, ferulic acid, Hep-2 cell, DNA fragmentation, oxidative stress