كفاءة بعض المركبات الطبيعية المعزولة والمشخصة من بعض النباتات العراقية وتقييمها الجزيئي كمضادات أكسدة وسرطان

الباحث

ابتسام ناظم حازم العساف

المشرف

مثنى جاسم الطائي - فواز فاضل علي

الجامعة

جامعة الموصل

الكلية / القسم

كلية التربية للعلوم الصرفة / قسم علوم الحياة

الشهادة

دكتوراه

السنة

2023

لغة الرسالة

عربي

الكلمات المفتاحية

النباتات الطبية العراقية ; المركبات الطبيعية ; مضادات الأكسدة ; مضادات السرطان

This study investigates the effectiveness of secondary metabolite compounds of three plants present in the Iraqi environment, namely the fruits of the fire thistle plant (Pyracantha coccinea), the fruits of the puncture vine (Tribulus terrestris), and the flowers of the wormwood (Artemisia absinthium). The study included two main axes: The first axis was completed in the Natural Products Laboratory, Department of Biology, College of Education for Pure Science, University of Mosul, Iraq, These experiments included the separation and identification of some active compounds from the three selected plants using the continuous extraction device soxhlet by following the successive solvent system hexane, ethyl acetate, and ethanol, then the extracts were concentrated using a rotary evaporator device to obtain the concentrated crude extract The fatty acids in the hexane extract were identified using Gas Chromatography–Mass Spectrometry (GC-MS) for the three plants, palmitic acid, stearic acid, Tridecanoic acid, and Hentriacontane, beta-Tocopherol, gamma-tocopherol, Arachidonic acid, Lauric acid, cis-Vaccenic acid, gamma-Sitosterol, the extracts were fractionated using CC Column Chromatography, and several phenolic compounds were identified using High-Performance Liquid Chromatography (HPLC) and the compounds in the fruits of the thistle plant, which are gallic acid, salicylic acid, catechol, 4- hydroxy benzoic acid, vanillic acid, and hydroquinone, in addition to the mentioned compounds, coumarin was detected and identified in the fruits of the elderberry plant, and the compound resorcinol in the flowers of the wormwood plant. The second axis, it was carried out outside Iraq at Atlas Biotechnologies Company in the Republic of Turkey, and it included evaluating the biological effectiveness of the separated compounds from the three plants against oxidative stress and cancer. The viability of treated cancer cells and cells control (breast cancer lines MCF7 and colon cancer HT29 compared with the healthy cell line WI38) with the plant extracts of the three plants was evaluated using the XTT test at concentrations of 10, 5, and 20 μg/ml. The treated breast cancer showed a significant reduction in cells viability compared with cell control with a ratio of 38.72, 39.54, and 33.26, respectively, while the same extract also affected the vitality of the colon cancer cell line with at a ratio of 31.78, 41.01, and 30.53, respectively with no significant effect on the healthy cell line. The effect of plant extracts was increased by increasing the extract concentrations for 24 hours of incubation. Furthermore, cells that experienced programmed death were significantly observed. The most common morphological changes in cell cultures of MCF7 and HT29 cancer cell lines included cellular changes represented by chromatin condensation, cell detachment and a change in shape to a circular shape, the presence of vacuoles, shrinkage of the cytoplasm, a decrease in the cells number and cells vitality, loss of the normal shape of the cells followed by the fragmentation of the nucleus into separate parts by budding. The cell, as a whole, produces membrane-bound apoptotic bodies, which means suppressing and inhibiting cancer with the concentrations used. The hallmark of dead and dying cells is the breakdown of cellular and nuclear membranes. However, no obvious changes were observed in the healthy cell line WI38. The results of the TUNEL test when using the three plant extracts at a concentration of 20 μg / ml and an incubation period of 24 hours showed the appearance of apoptosis features and antiproliferative activity of the three extracts. MCF7 breast cancer cell lines and HT29 colon cancer cell lines were determined cells that suffered from apoptosis stained with high fluorescent intensity by Andy Fluor 488 dye and fluoresced in green. The anti-Hoechst 33342 dye was used to identify cells that did not suffer from programmed death, which fluoresced in blue under a fluorescent microscope. The expression of the pro-apoptotic gene BAX and TP53 was measured after using the plant extracts. Gene expression analysis using RT- qPCR showed a significant increase in the gene expression activity of BAX and TP53 in breast and colon cancer cells compared to healthy cells at a concentration of 20 μg/ml of ethanolic extract. While the ethyl acetate extract of the fruits of the fire thistle plant gave the least amount compared to the control. The results of the gene expression of the antioxidant NRF2 in the cell cytoplasm when the plant extracts of the three plants were used on the breast and colon cancer cell line after 24 hours of incubation showed an increase in the amount of gene expression of the ethanolic extract of the plants of wormwood flowers, while the amount of gene expression of the two hexane and ethyl acetate extracts of the flowers of the plant decreased. Absinthe and elderberry fruits were also compared to control cells, and this indicates the ability of extracts to scavenge free radicals. The expression of CDK6 BRCA1and KRAS cell cycle regulator genes was measured after using the plant extracts. Gene expression analysis using RT-qPCR showed a significant decrease in their gene expression activity in breast and colon cancer cells compared to healthy cells at a concentration of 20 μg/ml of the ethanolic extract compared to control. --- page 631 [OCR+] --- Efficacy of Some Isolated and Diagnosed Natural Compound From Some Iraqi Plants and Their Molecular Evaluation As Antioxidant and Anticancer A Thesis Submitted by Ibtisam Nadhim Hazim AL-Assaf To The Council of College of Education For Pure Science University of Mosul In Partial Fulfillment of Requirements For the Degree of Ph.D. In Biology Supervised By Prof. Assist. Prof. Dr. Muthana Jasim AL-Taee Dr. Fawaz Fadhil Ali A.H. A.D. [OCR of page] Efficacy of Some Isolated and Diagnosed Natural Compound From Some Iraqi Plants and Their Molecular Evaluation As Antioxidant and Anticancer A Thesis Submitted by Ibtisam Nadhim Hazim AL-Assaf To The Council of College of Education For Pure Science University of Mosul In Partial Fulfillment of Requirements For the Degree of Ph.D. In Biology Supervised By Prof. Assist. Prof. Dr. Muthana Jasim AL-Taee — Dr. Fawaz Fadhil Ali 2023A.D. 1444 A.H. --- page 632 [OCR+] --- Efficacy of Some Isolated and Diagnosed Natural Compound From Some Iraqi Plants and Their Molecular Evaluation As Antioxidant and Anticancer Ibtisam Nadhim Hazim AL-Assaf Ph.D. Thesis Biology Supervised by Prof. Assist. Prof. Dr. Muthana Jasim AL-Taee Dr. Fawaz Fadhil Ali A.D. 1444 A.H. University of Mosul College of Education for Pure Science [OCR of page] 4A» University of Mosul | | College of Education for Pure Science Efficacy of Some Isolated and Diagnosed Natural Compound From Some Iraqi Plants and Their Molecular Evaluation As Antioxidant and Anticancer Ibtisam Nadhim Hazim AL-Assaf Ph.D. Thesis Biology Supervised by Prof. Assist. Prof. Dr. Muthana Jasim AL-Taee Dr. Fawaz Fadhil Ali 2023A.D. 1444 AH.

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