Narges Najjarpour, Masoud Mashadi Akbar Boujar,
Volume 3, Issue 1 (6-2016)
Abstract
Carnosic acid and Rosmarinic acid are family of polyphenols that are found in Rosmary plant. They have property biological behaviors such as anti-cancer, anti-viral and anti-oxidants. This study compared the effects of these two compounds based on ceramide metabolism in cell line of Hep- G2. In this experimental study, Hep-G2 cells were cultured in DMEM supplemented containing bovine fetal serum and antibiotics. Cells with double dilution were then cultured from tow substance Carnosic acid and Rosmarinic acid from concentrations 0 to 70 µM for 24 h and viability of cells was determined by MTT method. Spectrophotometer was used to measure of caspase-3 activity.To measure the level of ceramide, a recombinant ceramidase acid enzyme and naphthalene-2,3-dialdehyde, which is fluorescent and is connected to sphingosine resulted from ceramidase acid, were added to the cell extract and was ultimately determined by HPLC. In this study also activity of ensymes as sphingomylinase, acid ceramidase, glucosyl ceramide synthase was measured. Carnosic acid increased cell viability dose-dependent in Hep-G2 cells by reducing ceramide levels and decreasing activity of ensymes as caspse-3, sphingomylinase, glucosyl ceramide synthase and increasing activity of the enzyme acid ceramidase. Rosmarinic acid in concentrations of up to 50 µM decreased cell viability by increasing ceramide levels and activity of caspse-3, sphingomylinase and decreasing activity of enzymes as acid ceramidese and glucosyl ceramide synthase. Also this substance in concentrations of up to 40 µM caused increasing activity of caspse-3 enzyme. Although in most cases, polyphenols have resulted in induction of apoptosis and decreased cell viability, but in some cases they have inversely affected and caused cell growth.
Azra Saboora, Maryam Amiri Rad, Ezat Asgarani, Tayebeh Radjabian,
Volume 5, Issue 4 (12-2018)
Abstract
DNA extraction from plant tissues often causes most problems. For example, unsuccessful removal secondary metabolites during extraction, such as phenolic compounds in aromatic and medicinal plants, cause to some mistakes in result of molecular experiments by using of the extracted DNA. Achillea wilhelmsii is a medicinal plant belong to Asteraceae family and native to Iran, there is little information about genomic data in this plant. Therefore, optimizing of the DNA extraction methods for obtaining suitable quality and quantity yield is necessary. In this study, two traditional DNA extraction methods (using fresh and herbarium leaf samples) and commercial DNA kit MAGNANTM in Yarrow have been compared. Results showed that DNA extracted from fresh leaves of yarrow according to Khanuja et al. (1999) was better than the other methods which mentioned in this research, because of the increased amount of extracted DNA and reduced harmful compounds such as RNA, polysaccharides, protein and secondary metabolites. To evaluate the quality of extracted DNA from herbarium specimens showed that, in spite of the high yielding DNA (10-50 fold ratio to the other methods), quality of this extract was low on agarose gel because of smear and broken down the molecules. It can be considered as a limitation of the extract in molecular experiments. Procedure of MAGNANTM DNA kit was appropriate for reducing time and cost of the extraction as well as low contamination to protein and RNA, but the process needs some modifications for yarrow to increase the amount of extracted DNA.