Rameshi, N. (2016) Evaluation of immounomedulatory effects of bitter gourd fruit aquatic extract on human peripheral blood mononuclear cells. Masters thesis, university of zabol.
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Abstract
Biotechnology tools and their techniques provide new research to understand how healthy organs function. Understanding the molecular basis of health and disease leads to the discovery of new methods for treating and preventing the disease. Today, extensive studies have been conducted on the identification of modulators of immune response to natural compounds. One of these compounds is the extracts from the Karla or a bitter melon. This plant is known as "scientific name" Momordica charantia belongs to the cucurbitaceae family, which is traditionally used both as food and medicine. It is along with different properties such as anti-inflammatory, anti-diabetes, anti- Leukemia, anti-tumor, anti-mutation, antiviral, antimicrobial, etc. as well, it plays another role such as laxation of the body's immune system. The aim of this study was to investigate the immunomodulatory effects of its aqueous extract on the human peripheral blood mononuclear normal cells in the in-vitro conditions. The seed of the plant was cultivated in the greenhouse and its fruit was dried in dark green to light one green in shade and its powder was extracted using a water solvent by maturation method. The antioxidant capacity of the extract was determined in seven concentrations (30, 45, 60, 125, 250, 500, 1000 μg/ml) by DPPH method and its IC50 was determined. In addition, the protective effect of the extract (anti-genotoxicity, anti-cytotoxicity, anti-apoptosis), side effects of the extract (toxicity), as well as the expression of cytokines TNF-alph in the peripheral blood mononuclear normal cells were investigated in a randomized block design with three replications. These experiments were performed using 4 techniques such as chromosomal abnormalities (studying the cell genotoxicity), trypan blue coloring method (studying the cell cytotoxicity), DAPI coloring method (studying the cell apoptosis), Real Time PCR Technique (measuring the expression of TNF-alph cytokine). In all of these experiments, three concentrations of aqueous extract (60, 125 and 250μg/ml) were used for 24 hours and 48 hours on single-core cells under in-vitro conditions. To investigate the productive effect of extract, mitomycin C (dose: 0.25 μg/ml) for induction of cytotoxicity was used. The results showed that by increasing the concentration of aqueous extract of Carla, its antioxidant capacity will increase, and IC50 extract will be 108.05 μg/ml compared to positive control of BHA (IC50=30.90). Results of analysis of variance and mean comparison indicated that the extract alone in the studied concentrations is not along with the toxicity (genotoxicity, cytotoxicity ) effect on the normal cells, but also it will be decreased by increasing the concentration of genotoxicity extract and cytotoxicity was decreased at a concentration of 250 μg/ml (P<0.01), but did not have a significant effect on apoptosis and anti-apoptosis (P>0.05). On the other hand, by increasing the extract concentration, the protective effect of the aqueous extract of fruit (antigenotoxicity at a concentration of 125 μg/ml) and (P<0.01), anti-cytotoxicity at a concentration of 250 μg (P<0.01) and anti-apoptosis at a concentration of 125 μg/ml and (P<0.05) against mitomycin-C toxicity will be increased. By increasing the concentration of the extract after 24 hours, the expression of TNF-alph decreased (P<0.01), and the interaction of extract and mitomycin C significantly reduced the expression of TNF-alph (P<0.01). These findings, by illuminating Carla's immune modulatory properties, provide new perspectives for the production of effective therapeutic compounds.
| Item Type: | Thesis (Masters) |
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| Uncontrolled Keywords: | Carla Extract, Side Effects, Protective Effects, Mitomycin C, TNF-alpha |
| Subjects: | S Agriculture > SB Plant culture |
| Depositing User: | Mrs najmeh khajeh |
| Date Deposited: | 04 Dec 2018 09:21 |
| Last Modified: | 03 Mar 2019 08:07 |
| URI: | http://eprints.uoz.ac.ir/id/eprint/2315 |
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