Preetham Jinadatta
1*, Sharath Rajshekarappa
2, Kiran Sundera Raja Rao
1, Sujan Ganapathy Pasura Subbaiah
3, Sudhesh Shastri
41 Department of Biotechnology, Dayananda Sagar College of Engineering, Kumaraswamy Layout-Bangalore-560078, Karnataka, India
2 Department of Food Technology, Davangere University, Shivagangotri, Davangere -577007, Karnataka, India
3 Research and Development Centre, Nutrinorm Wellness Private Limited, Sahakar Nagar, Byatarayanapura, Bengaluru, Karnataka 560092, India
4 Department of Biotechnology, Kuvempu University, Jnanasahyadri, Shankaraghatta, Shimoga, Karnataka -57745, India
Abstract
Introduction: Gnetum ula is a notable medicinal plant used to cure various ailments. The stem part of the plant is used traditionally to treat jaundice and other disorders. The present work is to investigate the in vitro hepatoprotective and antioxidant activity of ethanol extract of stem of G. ula (GUE) and its isolated compound gnetol.
Methods: Column chromatography was carried out for GUE and various column fractions were obtained. DPPH and reducing power assays were performed for GUE and column fractions. The potent fraction was characterized, interpreted and tested for in vitro hepatoprotective activity on the BRL3A cell line. In silico docking studies of gnetol compound on the protein TGF-β (transforming growth factor – β) and Peroxisome proliferator-activated receptor α (PPARα) was carried out.
Results: DPPH scavenging and reducing power assay showed that the fourth column fraction has antioxidant potential than other fractions. The fourth column fraction was characterized to obtain gnetol compound. BRL3A cell line was used for the toxicity study of GUE and gnetol. Both, the extract and the isolated compound were found to be nontoxic with CTC50 value more than 1000 µg/mL. At the concentration of 200 µg/mL, GUE and gnetol offered cell protection of 50.2% and 54.3%, however, silymarin showed 77.15% protection at 200 µg/mL concentration against CCl4 treated BRL3A cell line. The docking results of the ligand molecule TGF-β showed that gnetol has the binding affinity of -7.0 and standard silymarin being -6.8. TGF-β showed good hydrophobic interactions and formed two hydrogen bonds with the amino acids. For PPARα protein, gnetol showed the binding affinity of -8.4 and silymarin with -6.5. Hydrogen bonding and good hydrophobic interactions against the amino acid molecules in relation to the PPARα protein are shown.
Conclusion: Gnetum ula stem extract and its isolated compound are safe and offered significant hepatoprotection against CCl4 induced toxicity. Isolated compound gnetol exhibited a potent antioxidant activity offering protection to liver damage. However, in vivo studies need to be carried out to validate the traditional use of G. ula.