Assessment of PunicagranatumL. extract on the mitotic arrest of plant bioassay system.

Abstract

Analysis of mitotic process and description of chromosome aberrations are important aspects in plant genetics to assess the effect of medicinal plant extract. Ethnopharmacological relevance: P. granatum extracts have been reported to exert multiple health effects, e.g. anti-inflammatory, antibacterial, and antidiarrheal, etc. However, the antiproliferative effects were not as extensively studied, including those of protocatachuic acid, a major constituent of the extract. This work aims at analyzing the constituents of the acetone extract of the plant and testing its antiproliferative effects. Material and Methods: Fingerprint of Punica granatum L. polyphenol acetone extract was analyzed using HPLC, UV-detection. Allium cepa L. root meristem was used as bioassay system. The study groups included 3 concentrations of the extract (0.1, 0.5, 0.75/ 100 mL) with 3 exposure times for each (3, 6 and 12 hours) and control for each treatment. For every experiment, 5 samples with 1,000 cells were studied. Effects on cytological and ultrastructure assays were done. As well as biochemical and molecular analyses (estimation of total protein content, DNA ladder, detection of nuclease activity) were done. Results: HPLC authentication of the extract revealed the presence of 15 fractions, of which protocatechuic acid, O-H Benzoic, chlorogenic acid, phenol and quarecetin were quantified. The application of acetone extract resulted in changes in ultrastructure of all organelles, reverse the viability of meristem cells and induce programmed cell death. The inhibitory effect of acetone extract was found to be dose and exposure time dependent. The mitotic process was associated with metaphase arrest and the disappearance of telophase stage that abolish the continuity of mitotic activity. There was a significant increase in nuclease activity enzymes, which in turn enhanced the cleavage of DNA to induce a typical ladder form. Conclusion: The total polyphenol extract of Punica granatum L. reprogram Allium cepa L. meristematic cells resulting in programmed cell death.