It may be said that LAECV contains mitostatic and metaphase arresting components that are able to induce significant metaphase arrest in root apical meristems and also in mouse bone marrow cells.
Clerodin (C24H34O7), a clerodane diterpenoid, is a bitter principle of Clerodendrum viscosum. The present study aimed to decipher colchicine-like actions of clerodin in terms of microtubule destabilization based mitotic abnormalities, G2-M arrest, and plant polyploidy. Purified clerodin showed increased metaphase frequency in Human Peripheral Blood Lymphocytes (HPBLs), Human Embryonic Kidney cells (HEK-293), and Allium cepa root apical meristem cells. Both squashed slide of the onion root tip and flow cytometric analysis of radish protoplast revealed a significantly increased frequency of polyploid cells. Flow cytometric analysis showed an increase in frequencies of G2-M in MCF-7 cells from 6.10 to 16.25% after clerodin (200μg/mL) treatment for 24 h. Confocal microscopy imaging of tubulin in clerodin-treated MCF-7 cells revealed microtubule destabilization. Molecular docking and LIGPLOT analysis indicate that clerodin interact in the colchicine binding site, including, single hydrogen bond with Asn 101 of α-tubulin. In summary, our experimental data revealed that clerodin has metaphase arresting, microtubule destabilization, and polyploidy inducing ability similar to colchicine. Molecular docking analysis revealed for the first time that clerodin and colchicine interact at the common site of tubulin residue indicating a common mechanism of action. The results also indicate similar cytotoxic potentialities of both clerodin and colchicine even though they belong to different chemical groups. Thus, clerodin may be used in place of colchicine as a plant polyploidy inducing agent in plant breeding programs in Agriculture.
Clerodendrum viscosum is a traditionally used medicinal plant and the earlier reports indicate its leaf aqueous extract (LAECV) contains metaphase arresting, cell cycle delay, and mitotic abnormality inducing active principles. The present study aimed to isolate pro-metaphase arresting, polyploidy, micronuclei and mitotic abnormality inducing active principles of LAECV. The LAECV was successively fractionated as petroleum ether (PEF), chloroform (CHF), and ethyl acetate (EAF) fractions. All the extract fractions were tested for Allium cepa and Triticum aestivum root swelling and root growth inhibition analyses. The petroleum ether fraction was selected for further cytotoxicity analysis on A. cepa root tip cells and was processed for detection of the active principles through HPLC, LC-MS, GC-MS and IR analyses. The comparative seedlings' root growth and swelling patterns indicate the bioactive principles are effectively fractionated in PEF and GC-MS analysis revealed the presence of Clerodin (m/z 434.3), 15-hydroxy-14, 15-dihydroclerodin (m/z 452), 15-methoxy-14, 15-dihydroclerodin (m/z 466), and 14, 15-dihydroclerodin (m/z 436) with the retention time of 14.038, 14.103, 14.480 and 14.655 respectively. Thus the present study explores clerodane diterpenoids of LAECV as pro-metaphase arresting, polyploidy, micronuclei, and mitotic abnormality inducing active principles.
Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze a detailed cytotoxic effect of the nonpolar petroleum ether fraction (AQPEF) of leaf aqueous extract of C. viscosum Vent. (LAECV) in Allium cepa root tip cells. The LAECV was fractionated with petroleum ether and tested for A. cepa toxicity at early hours (2 and 4 h treatment) at concentrations 0, 0.050, 0.100, 0.150 and 0.200 mg mL-1. The highest aberrant cell percentage (10.45%) was scored from 0.150 mg mL-1 followed by0.100 mg mL-1 (8.75%) concentration at 4 h treated samples. The AQPEF treatment induced a significant (p<0.0001) increase in micronuclei frequency; 4.31, 5.08, 5.05 and 3.05% respectively at concentrations 0.50, 0.100, 0.150, and 0.200 mg mL-1. The highest polyploid frequency (20.14%) induced with 0.100 mg mL-1 of AQPEF at 16 h recovery. 0.150 mg mL-1 is the most effective concentration of AQPEF to decipher its activity similar to colchicine (0.150 mg mL-1). In summary, the present study indicates petroleum ether is suitable for extraction of the active phytochemicals of LAECV having cytotoxic effects on A. cepa root tip cells. The AQPEF has colchicine like micronuclei, polyploidy, and mitotic abnormality inducing potentials in A. cepa root apical meristem cells.
Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze cell cycle delay, pro-metaphase arrest, and c-metaphase inducing effects of the petroleum ether fraction (AQPEF) of leaf aqueous extract of C. viscosum (LAECV). The LAECV was fractionated with petroleum ether and its metaphase arrest, cell cycle delay, and c-metaphase inducing activities were tested on A. cepa root tip cells. The AQPEF induced cell cycle delay, and colchicine like metaphase, c-metaphase, in A. cepa root tip cells. Thus, the present study explores AQPEF as an active fraction of LAECV having metaphase arresting, cell cycle delay, and c-metaphase inducing potentials.
Clerodendrum viscosum is a traditionally used medicinal plant. The present study aimed to analyze a detailed cytotoxic effect of the non-polar petroleum ether fraction (AQPEF) of leaf aqueous extract of C. viscosum Vent. (LAECV) in Allium cepa root tip cells. The LAECV was fractionated with petroleum ether and tested for A. cepa toxicity. Micronuclei, polyploidy, and mitotic abnormalities were analyzed after AQPEF treatment. The AQPEF induced concentration-dependent increased mitotic abnormalities, micronuclei, and polyploid cell frequency in A. cepa root tip cells and that were comparable with the colchicine effects. Thus, the present study explores that petroleum ether is a suitable solvent for extraction of the active phytochemicals of LAECV; AQPEF having colchicine like micronuclei, polyploidy and mitotic abnormality inducing potentials in A. cepa root tip cells, indicating its perspective use in karyotyping, plant breeding programs, and cytotoxicity studies.
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