Update: 2025-09-23
Shahla Hosseini
Faculty of Science / Department of Biology
Master Theses
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A Morphological and Micromorphological Study of Pollen Grains in Species of the Genus Salvia L. (Lamiaceae) with Emphasis on Species from Kurdistan Province
2025Botanical and morphological studies, particularly in the genus Salvia, play a significant role in the identification, classification, and understanding of the evolutionary relationships of plant species. In this research, the morphological diversity of pollen grains and the morphological characteristics of various Salvia species in Kurdistan Province, Iran, were studied. This province, due to its unique geographical and climatic conditions, provides an ideal habitat for various species of this genus, many of which possess medicinal and ecological properties. Specifically, the morphological characteristics of pollen grains from different Salvia species were examined using light microscopy. In this study, various Salvia species from Kurdistan were identified, and their morphological characteristics were analyzed using both qualitative and quantitative data. Cluster analysis based on qualitative and quantitative traits revealed that the species were grouped into diverse clusters and subclusters. The analyses based on qualitative traits identified variations in flower color, inflorescence type, and stem and leaf characteristics. Additionally, cluster analysis based on quantitative traits such as stem length, calyx length, and number of stamens revealed evolutionary relationships among the species. The results indicated that the clustering based on both qualitative and quantitative traits did not align with previous classifications in the Flora of Iran, and some species were placed into new groups. These discrepancies highlight the need for a revision of past classifications and further studies on the evolution and phylogeny of these species. The findings of this study could contribute to a better understanding of the environmental adaptations and biological differentiation of Salvia species in Kurdistan Province and serve as a basis for future research in the conservation and utilization of these valuable plants. Furthermore, this study also carried out clustering of Salvia species based on the micro-morphology of pollen grains using light microscopy. The results showed that the species were distinctly clustered based on pollen grain characteristics, such as size, shape, and surface features. These cluster analyses highlight prominent differences in the pollen grain structure of various Salvia species, which can be used as a criterion for species identification and differentiation in systematic and taxonomic studies.
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The impact of Ferula orientalis L. extract on rotenone-treated SH-SY5Y cell line
2024Nowadays, there is increasing attention to natural compounds with antioxidant properties in plants, as they generally have fewer side effects. Studies have been conducted on the compounds and biological effects of the species Ferula orientalis from the Apiaceae family, which was collected from the highlands of the Kosalan region in Kurdistan Province, and some of its biological properties were examined. Parkinson's disease (PD) is among the incurable and rapidly growing diseases in human societies. In Parkinson's, with the loss of neurons, especially in the substantia nigra of the brain, the patient faces irreversible motor and cognitive impairments. Neuronal damage and death are associated with accumulations of the alpha-synuclein protein. In this study, we observed that certain toxins, such as rotenone, can induce Parkinson’s by creating stress conditions on neuronal cells (SH-SY5Y). Rotenone acts as a neurotoxin, leading to the destruction of neuronal cells. This study investigated the antioxidant effects and potential of Ferula extract at various concentrations on Parkinsonian cells induced by rotenone using the MTT assay. Antioxidant capacity assessments of the extract were conducted through ferric reducing power (FRAP) and DPPH radical scavenging tests. Additionally, the total phenol and flavonoid content of the species was evaluated using the Folin-Ciocalteu method and the aluminum chloride colorimetric method, respectively. The chemical composition of the plant's essential oil was also analyzed using GC-MS. The results showed that this species has significant reducing power and radical scavenging activity, which may be attributed to its high phenol and flavonoid content, amounting to 30.82 mg of gallic acid per gram of extract and 15.96 mg of quercetin per gram of extract, respectively. However, the IC50 value of 4232 µg/mL for DPPH radical scavenging indicates a relatively low antioxidant potential compared to the Trolox standard. At the tested concentrations (62.5 to 1000 µg/mL), the plant extract could not inhibit 50% of free radicals, and higher amounts of the extract would be required for inhibition. In assessing the survival rate of neuronal cells treated with rotenone and the extract through the MTT assay, it was found that treatment with the extract in all three conditions (treatment, pretreatment, and simultaneous) at relatively low extract concentrations (3 to 50 µg/mL) yielded favorable results. However, in the pretreatment and simultaneous conditions (particularly pretreatment), cell survival rates were higher than in the treatment condition, possibly due to the presence of antioxidants from the extract before rotenone, which could neutralize the free radicals generated by rotenone before causing cellular damage. At higher extract concentrations (100 and 150 µg/mL), in all conditions, cell survival rates decreased. Based on the obtained results, it can be concluded that Ferula orientalis can reduce oxidative stress, one of the main causes of Parkinson's, and prevent further cell destruction. This plant can be introduced as an effective medicinal herb with high antioxidant properties and may also serve as a potent anti-cancer agent by inhibiting the growth and proliferation of cancer cells.
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The effect of the extract and essential oil of Thymus kotschyanus Boiss. et Hohen on the viability and morphology of the cellular model of Parkinson's disease
2024From ancient times to the present, medicinal plants have been utilized in various aspects. The genus Thymus, belonging to the Lamiaceae family, comprises 928 species worldwide, with 18 aromatic species found in Iran. Thymus kotschyanus has numerous medicinal and culinary uses. In this study, ecotypes of this species were collected from the Kousalan region of Kurdistan province, and some of its biological properties were examined. Parkinson's disease is the second most prevalent neurodegenerative disorder, characterized by the loss of dopaminergic neurons in the substantia nigra of the brain. Various genetic and environmental factors contribute to its pathogenesis, including oxidative stress. Oxidative stress can lead to mitochondrial dysfunction, disrupting its function and increasing apoptosis. Rotenone, a type of pesticide, can have adverse effects on human health by inhibiting Complex I of the electron transport chain, leading to mitochondrial dysfunction, increased oxidative stress, and apoptosis. This study investigated the antioxidant potential of Thymus kotschyanus extract and essential oil in enhancing the viability of a Parkinson's cellular model induced by rotenone using the MTT assay and immunocytochemistry. Additionally, the antibacterial potential of the extract and essential oil was evaluated against Bacillus subtilis and Salmonella typhi using the disk diffusion method and the tube dilution method to determine MIC and MBC. It is worth mentioning that evaluations of the antioxidant potential of the extract and essential oil were conducted using methods such as iron reduction and DPPH radical scavenging assays. Furthermore, the total phenolic and flavonoid contents of the studied species were assessed using the Folin-Ciocalteu and aluminum chloride colorimetric methods. The chemical composition of Thymus kotschyanus essential oil was also analyzed using GC-MS. The results indicated significant radical scavenging and inhibitory activities of this species, likely due to its high phenolic and flavonoid content. Although there was a significant difference compared to ascorbic acid and trolox standards, the IC50 of 557 µg/ml for DPPH radical inhibition demonstrated the high antioxidant potential of this species. The phenolic and flavonoid contents were 25.38 mg GAE/g and 58.91 mg QUE/g, respectively. The extract inhibited the growth of both Gram-positive (B. subtilis) and Gram-negative (S. typhi) bacteria, with MIC and MBC values of 100 and 200 mg/ml, respectively. The essential oil was effective at all concentrations against both strains, with the highest inhibition zone at the highest concentration (300 µl/ml) exceeding that of the antibiotic (positive control). The results of the tube dilution test showed that the extract had MIC and MBC values of 0.78 and 1.56 µl/ml for B. subtilis and 0.39 and 0.78 µl/ml for S. typhi, respectively, indicating high inhibitory and bactericidal activity of the studied species at low concentrations for both strains. Regarding the MTT assay, the essential oil was unable to reduce rotenone-induced toxicity and, conversely, acted as a factor in decreasing cell viability. Different concentrations of the essential oil showed that pre-treatment with the oil resulted in the highest cell viability, while post-rotenone treatment resulted in the lowest cell viability. Similarly, the extract showed the highest cell viability when treated before rotenone exposure, and the lowest viability post-rotenone treatment. The results of the immunocytochemistry assay demonstrated that the expression of the gene or the appearance of tyrosine hydroxylase protein increased at a concentration of 50 µg/ml, indicating an increase in dopamine precursor in the Parkinson's cellular model under study. The obtained results lead to the conclusion that ecotypes of the T. kotschyanus species from the Kousalan region of Kurdistan province can be introduced as an effective medicinal plant, possessing high antioxidant and antibacterial properties. Additionally, this plant, with its significant inhibitory effect on the growth of prokaryotic cells, can be a suitable option for inhibiting the growth and proliferation of cancer cells as well.
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Phytochemical Analysis and Biological Properties of Nanoessential Oil from Achillea Biebersteinii Afan in Kurdistan Province
2024The Asteraceae family comprises numerous flowering plants, consisting of 1600 genera and 23000 species worldwide. The species Achillea Biebersteinii has been utilized for centuries in traditional medicine globally, possessing properties such as pain relief and wound healing, and has been employed as an herbal remedy in traditional medicine. This article investigates the phytochemical properties of the extract of A. Biebersteinii species through tests including assessment of its antioxidant potential using DPPH and FRAP methods, evaluation of phenolic and flavonoid contents, as well as analysis of compounds using Gas Chromatography/Mass Spectrometry (GC/MS). Additionally, after preparing nanoemulsions via the emulsion phase inversion method, the antimicrobial properties of the extract and essential oil, along with the produced nanoemulsions, were compared through tests including the zone of inhibition assay, biofilm formation inhibition assay, antibiotic synergy test, and determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The results of the FRAP and DPPH antioxidant tests demonstrate the extract's good iron reduction and radical scavenging abilities. However, significant differences were observed compared to standards such as ascorbic acid and Trolox. The effective concentration for DPPH radical inhibition (IC50=964μg/ml) was determined. Although the extract was incapable of reducing 50% of Fe3+, the calculation of its EC50 was therefore disregarded. Furthermore, the results of total phenolic and flavonoid content analysis of the studied species indicated the presence of 66.30 mg gallic acid and 55.41 mg quercetin per gram of extract, respectively. Gas chromatography-mass spectrometry (GC-MS) analysis of the essential oil of this species revealed 58 compounds, with the highest volume percentage belonging to monoterpenoids (such as camphor, eucalyptol), terpinols, and aromatic hydrocarbons, which are utilized in food, cosmetic, hygiene, and pharmaceutical industries. Based on the results from the biofilm formation inhibition assay, nanoemulsions prepared from the essential oil exhibited better efficacy compared to natural oil. Nanoessential oil of this species showed more significant inhibition of biofilm formation in the fungal pathogen Candida albicans compared to bacterial strains Eschrichia coli and Staphylococcus aureus. The extract showed positive effects against E. coli in inhibiting biofilm formation. According to the results of the antibiogram test, based on the zone of inhibition, the plant extract exhibited significant effectiveness against C. albicans at both concentrations (100 and 300 micrograms per milliliter). The extract also inhibited the growth of S. aureus. Nanoemulsions also showed positive effects at higher concentrations against bacterial strains Bacillus cereus and S. aureus. However, no significant results were observed for antibiotic synergy tests and MIC/MBC assays. This implies that microbial growth was observed at all concentrations of the plant extract used, and no enhancement in microbial proliferation inhibition was observed in any concentration of the extract combined with antibiotics. Nevertheless, due to its notable antioxidant property and inhibition of growth and proliferation of fungal pathogen C. albicans and bacterial strains such as Bacillus cereus and S. aureus, it could be utilized, provided the preservation of its species, in pharmaceutical and food industries.
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Investigation of phytochemistry and cytotoxic effect of Allium saralicum Fritsch R.M. extract on HeLa cancer cells
2024The genus Allium belongs to the family Amaryllidaceae (the Amaryllis family) of the flowering plant order and is part of the monocotyledonous group, one of the largest families in this group. This extensive taxon includes approximately 918 heterogeneous species worldwide, which are native to dry and temperate regions of the Northern Hemisphere. Allium saralicum, with a regional distribution in Iran, Turkey, and Iraq, is one of the species that has been reported for its medicinal properties in numerous studies. The aim of this research was to study ecotypes from the Saral region of Kurdistan province and to investigate the phytochemistry of various aerial parts (stem, leaf, and flower) of this species, as well as to examine the cytotoxic effect of ethanol and methanol extracts of these parts on HeLa cancer cells. Antioxidant capacity of the species was assessed using FRAP and DPPH methods, phenol and flavonoid content were measured using Folin-Ciocalteu and aluminum chloride colorimetric methods respectively, and cellular toxicity was evaluated using the MTT assay. Furthermore, the chemical compounds present in the methanol extract of aerial parts of A. saralicum were analyzed using GC method. FRAP test results indicated that methanol leaf extract of A. saralicum exhibited the highest reducing power compared to other samples, while the lowest reducing power was attributed to ethanol flower extract of the same species. In DPPH antioxidant assay, ethanol leaf extract showed the highest radical scavenging activity, whereas ethanol flower extract of A. saralicum exhibited the lowest activity, similar to the FRAP test results. The lowest concentration required to inhibit 30% of radicals (IC30) was obtained for ethanol leaf extract (171.68 µg/ml), while the highest belonged to ethanol flower extract (1299.42 µg/ml). In terms of phenolic content, methanol leaf extract had the highest amount (233409.0 ± 42273.17 mg AG/gr extract), while ethanol leaf extract had the lowest (50593.14 ± 275505.0 mg AG/gr extract). Ethanol leaf extract also showed the highest flavonoid content (9846.3 ± 327.37 mg Quercetin/gr extract), while ethanol flower extract had the lowest (50593.9 ± 09833.0 mg Quercetin/gr extract). Chemical analysis of methanol extract of all aerial parts of A. saralicum revealed three compounds with high percentages. The highest percentage was attributed to Methyl 2-cyano-3,4-Dimethyl-4-phenylcrotonate, which is an ester group compound, and two other compounds were examined in chemical databases but no similar compounds were found. In MTT cytotoxicity assay, all samples demonstrated significant effects on the viability of cancer cells at various concentrations, even at low concentrations, cell viability decreased. Based on the results obtained, A. saralicum can be introduced as a medicinal species in the province, and by paying attention to the conservation of its biodiversity, it can be introduced to the pharmaceutical industry.
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Investigation of phytochemical properties and determination of synergistic power of Arum sp. and Eminium intortum (Banks & Sol.) Kuntze extracts from Araceae with common antibiotics
2023Araceae is a monocotyledonous plant family that includes approximately 144 genera and over 3645 species divided into numerous subfamilies. Several Araceae species have been cultivated by humans for a variety of purposes ranging from food to medicine. The current study used dried leaves and flowers of the chosen plant species that were obtained from the Kurdistan region of Iraq and the Kosalan mountain region of Iran to assess the antioxidant, antibacterial, and synergistic effects of certain Araceae. For this study, two Araceae species—Arum conophalloids L. and Eminium intortum (Bloome) schott—were selected. After drying for one to two weeks at room temperature in the shade and out of the sun, methanolic extracts of a given species were prepared. The DPPH and FRAP tests were performed to evaluate the extracts' antioxidant activity. The Folin-Ciocalteau and aluminum chloride color complex techniques were used to measure the total phenolic and total flavonoid contents, respectively. The identification of chemical components and the produced metabolites was carried out using Gas Chromatography-Mass Spectroscopy (GC-MS) analysis. Additionally, a variety of antimicrobial tests, such as the agar well diffusion test, disc diffusion test, MIC, MBC, biofilm assay and synergistic effect with common antibiotics were conducted to determine their effect upon bacterial and fungal cells. The study's findings indicated that both Arum (leaf and flower) and Eminium (leaf and flower) extracts had low antioxidant capacities, Eminium intortum leaf extract had the highest antioxidant capacity belongs to the Arum species and Eminium intortum flower extracts. The assessment of antioxidant capacity using the FRAP method with Reduction inhibition percent showed that E.intortum leaf had the highest reductive power and the DPPH test results, with their IC20 values showed that Eminium intortum flower extract is more effective in inhibiting free radicals compared to other extracts. Additionally, E.intortum leaf extract exhibited the highest flavonoid content with 16.55952±6.101998 milligrams of quercetin per gram of extract. Although A.conophalloides leaf allocated the highest phenolic content to itself with 19.32927±1.24647 milligrams of gallic acid per gram of extract. The bioactive compounds found in Arum conophalloides leaf extract were identified by GC-MS analysis. These included cadin-3,5-diene (a sesquiterpenoids), Vitamin A, 6-Aza-5,7,12,14-tetrathiapentacene (alkaloids) and (steroids, monoterpenoids) Cyclopentane, 1,2-dimethyl-3-(1-methylethenyl). Other bioactive compounds found in Eminium intortum leaf included 2,6,10-TRIMETHYL,14-ETHYLENE-14-PENTADECNE (terpenoids), Phthalic acid esters (dialkyl), 4,7-Diphenyl-9-dimethylamino-5H-dibenzo[c,e]azepin (Benzazepines), and 3-METHYL-5-DIPHENYLDIHYDRAFURAN (Fatty Acids).Additionally, Antimicrobial tests indicated that Eminium intortum flower extract had greatest inhibitory effects on the candida albicans with low concentration of extract but does not show any inhibitory effect on the tested bacteria with the same concentration, while at higher concentration 100mg/ml the extracts have a few effects on Bacillus cereus, Staphylococcus aureus and Escherichia coli was approximately ineffective, among the studied bacteria, all extracts showed the higher ability to inhibit bacteria growth against Bacillus subtilis and they are don’t have any inhibitory effect on Pseudomonas aeruginosa even at higher concentration. The MIC and MBC test performed for all extracts against Bacillus subtilis and candida albicans, only Arum conophaloids leaf extract make inhibition at 50mg/ml two-fold decrease on Bacillus subtilis.As well as the checkerboard test was showed that the effectiveness of various extracts in augmenting the potency of the commonly employed antibiotics, penicillin and Amphotericin B, against Bacillus cereus and Candida albicans. Following a 24-hour period, noteworthy enhancements were observed in the efficacy of both antibiotics for the specified microorganisms. biofilm formation assay tests were performed for extracts do not show antimicrobial activity, it produces a false positive inhibition in studies microbes due to the presence of small extract particles.
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Phytochemical investigation of the extract and essential oil of Thymus kotschyanus Boiss. (Lamiaceae) and its inhibitory effect on biofilm formation in gram positive and negative bacterial strains
2023In this research, the first step involved an examination of the biological properties of the extract from the species Pimpinella anthriscoides within the Umbelliferae family. The extract of this plant was assessed for its antioxidant properties using methods such as Ferric Reducing Antioxidant Power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, as well as the total phenolic and flavonoid content. Due to the absence of essential oil in this species and the extract's inability to inhibit the growth of target bacteria, the species Hymenocrater longiflorus from the Lamiaceae family was selected. Antioxidant tests, assessment of phenolic and flavonoid content, and various microbial evaluations were also conducted for the extract of this species. The amount of essential oil obtained from the aerial parts of this species was negligible, and it was insufficient for conducting additional microbial tests (biofilm inhibition, Minimum Inhibitory Concentration (MIC), and Minimum Bactericidal Concentration (MBC)). Therefore, another species from the Lamiaceae family, Thymus kotschyanus, was chosen for comparison with H. longiflorus, as it had a significantly higher essential oil yield. The biological properties of T. kotschyanus included an examination of its antioxidant capacity using FRAP and DPPH methods, phenolic content using the Folin-Ciocalteu method, flavonoid content using the aluminum chloride colorimetric method, and antimicrobial effects using various techniques (zone of inhibition using disk diffusion, tube dilution method for estimating MBC and MIC, and quantitative and qualitative assessment of biofilm inhibition) against the bacterial strains Escherichia coli and Staphylococcus aureus. The results of the FRAP assay showed that H. longiflorus exhibited higher reducing power at all concentrations compared to the other two plant species. Although the DPPH assay indicated that the extract and essential oil of T. kotschyanus were more effective in scavenging free radicals. Evaluation of phenolic and flavonoid content revealed that P. anthriscoides had the highest flavonoid content at 1/100 milligrams quercetin per gram of extract and the highest phenolic content at 36.65 milligrams gallic acid per gram of extract. However, the results of microbial tests demonstrated that even at the highest concentration, the extract of P. anthriscoides was ineffective in inhibiting the growth of the studied Gram-positive and Gram-negative bacterial strains. Therefore, further microbial tests with the presence of this species' extract were abandoned. H. longiflorus extract produced a growth inhibition zone with a diameter of 13 millimeters for S. aureus and 8 millimeters for E. coli at the highest concentration (300 mg/mlµ), which was acceptable compared to the other two species' extracts that were ineffective in inhibiting bacterial proliferation. The essential oil of H. longiflorus also generated a growth inhibition zone with a diameter of 7 millimeters against E. coli at the highest concentration (100 μl/mlµ). The essential oil of T. kotschyanus exhibited growth inhibition zones with diameters of 14 and 16 millimeters against E. coli, and 13 and 16 millimeters against S. aureus at concentrations of 25 and 50 microliters per milliliter of oil, respectively. At a concentration of 100 microliters per milliliter, this oil induced 100% killing for both bacterial strains, preventing their growth and proliferation to form biofilms. MIC tests for T. kotschyanus showed that at lower concentrations, the essential oil was capable of inhibiting bacterial proliferation (MIC=6.25 µl for S. aureus and MIC=12.5 µl for E. coli). Biofilm inhibition assessment also indicated that the essential oil of T. kotschyanus was effective at MIC/2 but showed no significant results at MIC/4 and MIC/8. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of the essential oil from T. kotschyanus revealed that its major component was Linalool (49.17%). The acceptable antioxidant and antimicrobial properties of the essential oil and extract from H. longiflorus and T. kotschyanus suggest their potential medicinal value.
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Evaluation of antioxidant power and phenolic content of of Viola odorata L. and Hymenocrater longiflorous Benth. extracts From Kosalan region of Kurdistan province
2023This research focuses on the study of two plant species, Viola odorata L. from the Violaceae family and Hymenocrater longiflorous benth. from the Lamiaceae family, as ecotypes from the Kosalan region of Kurdistan. Viola odorata is a member of the order Malpighiales within the family Violaceae, and it is native to Europe and Asia, representing the largest genus within the Violaceae family with approximately 600-525 species worldwide. In Iran, there are 98 species within this genus, with 83 of them being reported exclusively from Iran. Hymenocrater is a genus within the Lamiaceae family, comprising 89 species worldwide and having a distribution in Iran, Afghanistan, Turkmenistan, Pakistan, Turkey, and Iraq. Among these, 88 species have been reported in the Iranian flora, with four of them being endemics. The specimens for this study were collected from the Kosalan region of Kurdistan in the spring of 1402. This study primarily focuses on assessing the antioxidant properties using iron reduction and DPPH radical scavenging methods, determining the flavonoid and total phenol content, and conducting antimicrobial assays (antibiogram and minimum inhibitory concentration) on the mentioned plant species. The evaluation of antioxidant capacity using the iron reduction method revealed that the flower of H. longiflorus exhibited the highest reductive power. Furthermore, this species displayed better performance in scavenging free radicals, with the lowest IC50 (851.44 micrograms per milliliter) being closer to the inhibitory concentration compared to the standard Trolox (612.35 micrograms per milliliter). Additionally, the leaves of V. odorata had the highest flavonoid content at 21.58 milligrams of quercetin per gram of extract, while the flowers of this species had the lowest amount at 3.37 milligrams. The phenol content in the studied samples varied slightly, ranging from 30 to 33 milligrams of gallic acid per gram of extract. The results of the antimicrobial experiments indicated that the flower extract of V. odorata did not create sensitivity in both strains at different concentrations. However, the leaves of this species created a growth inhibition zone with a diameter of 7 millimeters for Escherichia coli at the highest concentration (200 milligrams per milliliter). Moreover, the leaves of H. longiflorus only induced a growth inhibition zone for Staphylococcus aureus at all concentrations and had no effect on Gram-negative bacteria. The flower of H. longiflorus displayed superior performance by creating a growth inhibition zone (13 millimeters) for S. aureus at all concentrations and exhibiting the best performance at a concentration of 200 milligrams per milliliter against E. coli compared to other samples. The results of the tube dilution assay indicated an MIC of 25 mg/ml and MBC of 50 mg/ml for the flower and leaves of H. longiflorus against S. aureus, and an MIC of 200 mg/ml against E. coli, with the flower extract of V. odorata showing no effect but the leaf extract confirming an MIC of 200 mg/ml. These findings were consistent with the antibiogram test.Gas chromatography analysis of the samples confirmed the presence of a variety of compounds, including terpenoids, sesquiterpenoids, alkaloids, sugars, esters, and more in the studied species. Based on the presence of these specific compounds, it is anticipated that these species may possess antidiabetic and anticancer properties. In conclusion, the data suggest that H. longiflorus could be introduced as a medicinal species in the future with appropriate conservation measures.
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Phytochemical screening and antimicrobial effects of extracts and essential oils of some species of Apiaceae family
2023The umbelliferous family, with around 440 genera and 3,500 species, is considered one of the largest plant families in the world. It has various applications in terms of consumption, medicinal use, cosmetics, and hygiene. Understanding the chemical compositions and biological effects of species within this family is crucial for advancing medical and pharmaceutical goals. This study focuses on investigating the essential oils and extracts of three species (Chaerophyllum macrospermum, Prangos ferulacea, Ferula orientalis) from the Kurdistan region. The biological properties of these target species were evaluated, including antioxidant capacity using FRAP and DPPH methods, phenolic and flavonoid content determination, by Folin–Ciocalteu and aluminum chloride colorimetry respectively, and antimicrobial effects against Escherichia coli and Staphylococcus aureus using various methods. Among the studied species, C. macrospermum was selected for its successful properties, and the anticancer effect of its essential oil was evaluated on HeLa cell lines using the MTT method. Probable chemical compounds in the essential oils were identified using GC-Ms. Evaluating antioxidant capacity using the FRAP method revealed that C. macrospermum and F. orientalis had the highest reductive power, with respective IC50 values of 854±1.51 μg/ml and 916±1.51 μg/ml in the DPPH assay. Moreover, C. macrospermum exhibited the highest flavonoid content at 1.3±31.28 mg quercetin per gram of extract, while P. ferulacea had the highest phenolic content at 0.38±30.82 mg gallic acid per gram of extract. Antimicrobial tests showed that P. ferulacea extracts and essential oil had no effect on the growth of Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria. However, C. macrospermum essential oil inhibited growth and induced lethality at concentrations of 75 and 150 microliters per milliliter. It also significantly affected biofilm formation, particularly in Gram-positive strains, even at concentrations below 75 microliters per milliliter. The cytotoxicity of C. macrospermum essential oil on HeLa cell lines was evaluated, demonstrating its anticancer potential even at low concentrations (18.75 μl/ml). Despite higher absorption due to partial insolubility in 1% DMSO as a solvent at higher concentrations (a pseudo-absorption effect), the images of cells indicated cytotoxic effects of the essential oil in all concentrations, confirming its positive impact. Chemical composition analysis of the studied essential oil showed that C. macrospermum had the highest percentages of ψ-Limonene, L-α-Pinene, and Ocimene-β; F. orientalis had the highest percentages of α-Pinene and Fenchyl acetate; and P. ferulacea contained predominant compounds like 3-Carene and Terpinene-ɣ. Based on the research findings, the studied species are rich sources of monoterpenes and esters, with C. macrospermum and F. orientalis showing high potential for pharmaceutical and food industries.
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مطالعه تنوع ژنتیکی تعدادی از گونه های جنس (Amaryllidaceae) Allium با استفاده از نشانگرهای ISSR
2022جنس Allium ، بهویژه زیر جنس Melanocrommyum شامل بخشهایی است که از لحاظ تاکسونومیکی بسیار پیچیده بوده و محتوای بخشهای آن در طول زمان به دفعات بازبینیشده است. در مطالعهی حاضر ارتباط بین 32 اکوتیپ متعلق به 10 گونه مختلف از جنس Allium، با استفاده از نشانگر ISSR مورد بررسی قرار گرفت. 9 آغازگر مورد استفاده، 166 باند چند شکل را تولید کردند (متوسط 18 باند به ازای هر آغازگر). از بین آغازگرهای مورد استفاده، آغازگر ISSR873، با 27 نوار، بیشترین و آغازگر ISSR4 با 2 نوار کمترین تعداد نوار چندشکل را ایجاد نمودند. PIC نشانگرها بین 04/0 تا 43/0 متغیر بود. تجزیه خوشهای به روش UPGMA و تجزیه به مختصات اصلی، اکوتیپ های موردمطالعه را در چهار گروه قرار داد. نتیجه خوشه بندی و تجزیه به مختصات اصلی نشان داد که بیشتر گونههایی که از لحاظ ریخت شناختی به هم شبیه هستند در گروههای نزدیک به هم قرار گرفتند. این مساله میتواند تک نیا بودن گونههای موردمطالعه را تایید کند. اگرچه اکوتیپی از گونه A. hooshidaryae نزدیک به گونه A. saralicum در بخش Melanocrommyum قرار گرفت که این احتمالاً میتواند با حوزه پراکنش جغرافیایی یکسان آنها توجیه شود. بر اساس ضریب تشابه Dice، بیشترین درصد تشابه در بین اکوتیپهای 4 A. stipitatum و A. saralicum 4 (72%) و کمترین شباهت در بین اکوتیپهای مربوط به گونههای A. tripedale و A. iranicum (12 %) برآورد شد. اکوتیپ های با کمترین درصد تشابه به زیر جنس های Allium و Nectaroscordum تعلق دارند که در خوشههای مجزا قرار گرفتند.
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اثر عصاره ی آبی-الکلی میوه گیاه بامیه (Abelmoschus esculentus) بر سمیت سلولی ناشی از استرپتوزوتوسین در رده ی سلولی بتای پانکراس
2022دیابت یکی از شایع ترین اختلالات غدد درون ریز است که بر دو نوع است، دیابت نوع اول ناشی از تخریب خود ایمنی سلول های β پانکراس است و معمولاً منجر به کمبود مطلق انسولین می شود. دیابت نوع دوم که شامل افرادی می شود که مقاومت به انسولین دارند. در این مطالعه از استرپتوزوتوسین به عنوان عامل القا کننده دیابت استفاده شد. دیابت باعث افزایش نیتریک اکساید و افزایش رادیکال های آزاد می شود. بسیاری از گیاهان، ادویه جات و سایر مواد گیاهی برای درمان دیابت در سراسر جهان معرفی شده است. ثابت شده است که بامیه (A.esculentus) می تواند به عنوان یک مکمل غذایی به منظور جلوگیری از عوارض ناشی از استرس اکسیداتیو در دیابت استفاده شود. در این تحقیق سلول های بتای پانکراس کشت داده شدند و سپس با استرپتوزوتوسین دیابتی شدند و نهایتا اثر عصاره گیاه بامیه روی این سلول ها بررسی شد. سنجش میزان ترشح انسولین به روش الایزا و کیت اختصاصی با آنتی بادی اختصاصی برای انسولین انسانی در پلیت های 96 خانه ای انجام شد. سلول ها پس از تیمار با STZ و عصاره با تریپان بلو مخلوط و روی لام نئوبار زیر میکروسکوپ مشاهده شدند. سپس سلول های زنده و مرده شمارش و درصد سلول های زنده محاسبه شد. از مایع رویی خانه های ظرف کشت همه ی گروه ها و کنترل جمع آوری و جهت سنجش نیتریک اکساید استفاده شد. ظرفیت تام آنتی اکسیدانی به روش فرپ انجام شد. سلول های رده ی بتای پانکراس تحت تاثیر عصاره و STZ قرار گرفته شد. STZ باعث مرگ سلولی و کاهش معنی دار در تعداد آنها شد. اما همراهی عصاره با STZ، نشان داد که عصاره سمیت سلولی STZ را کاهش داده و باعث افزایش معنی دار تعداد سلول ها می شود. همچنین مشخص شد که دیابت القا شده توسط STZ باعث کاهش معنی داری در ترشح انسولین و اثر عصاره همراه با STZ باعث افزایش معنی دار ترشح انسولین شد. عصاره به تنهایی تغییر چندانی در سطح نیتریک اکسید نداشت اما در سایر گروه های تحت تیمار عصاره و STZ در سطح نیتریک اکساید، افزایش معنی داری را نشان داد. اما در گروه STZ+Extract عصاره باعث کاهش سطح نیتریک اکسید شد. در نهایت STZ سبب افزایش معنی دار در سطح FRAP شد. ظرفیت تام آنتی اکسیدانی در تمام گروه ها نسبت به گروه کنترل افزایش پیدا کرد و معنی دار بود. در حالی که گروه تیمار شده با عصاره تغییر معناداری را در سطح FRAP نشان نداد و شبیه گروه کنترل بود.
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ارزیابی کروموزومی گونههایی از جنس آلیوم (نرگسیان) در منطقه سارال کردستان
2022جنس Allium متعلق به خانوادهی Amaryllidaceae (نرگسیان) میباشد، که شامل بیش از 1000 گونه دوساله یا چندساله، پیازدار و برخی دیگر دارای ریزوم یا ریشه ذخیرهای که دارای خواص دارویی و زینتی است. دانش سیتوژنتیک اثرات ژنتیکی، رفتار و ساختار کروموزومها را موردمطالعه قرار میدهد. مطالعات سیتوژنتیک دارای اهمیت زیادی است از جمله: برای شناخت ویژگیهای ژنتیکی گیاه (بومی، وحشی) و تشخیص وضعیت کروموزومهای گیاه است و با استفاده از اطلاعات کروموزومی مقایسهی گونهها و جمعیتها ممکن میشود. در مطالعه حاضر غدهی پیاز گونههایی از زیرجنس های Allium و Melanocrommyum از حوزههای مختلفی از منطقه سارال استان کردستان جمع آوری و مورد بررسی قرار گرفت. گونههای مورد مطالعه شامل: A.Saralicum،A. haemanthoides ، A. ubipetrense، A. stipitatum و A. ampeloprasum بودند. در این تحقیق ساختار و وضعیت کروموزومهای گونهها با استفاده از روش سیتوژنتیک کلاسیک (اسکواشینگ) مورد بررسی قرار گرفت. از نرمافزار ایدیوکار برای محاسبه پارامترهای کاریوتیپی و کروموزومی استفاده شد. از هر گونه حداقل 5 پلاک سلول متافازی مورد تجزیه و تحلیل قرار گرفت. نتایج نشان داد که بجز گونه A. ampeloprasum که تتراپلویید بود بقیه گونه ها دیپلوئید بوده و دارای عدد پایه کروموزومی 8x= هستند. در گونه A. saralicum یک جفت ریز ماهواره بر روی بازوهای کوچک کروموزوم ها و همچنین در گونه های A. Saralicum، A. haemanthoides و A. stipitatum 1-2 B کروموزوم مشاهده شد. در این مطالعه تصویر میکروسکوپی پلاک های کروموزومی متافازی، ایدیوگرام و مشخصات کاریوتیپ گونه های موردمطالعه ارائه شده است. لازم به ذکر است سیتوژنتیک گونه A. Saralicum برای اولین بار گزارش شده است. نتایج این پژوهش نشان داد که A. ubipetrense و A. stipitatumبه ترتیب متقارن ترین و نامتقارن ترین کروموزومها را دارند. فرمول کاریوتیپی گونه ها به شرح ذیل تعیین گردید: A. ubipetrense =16m A. saralicum= 14m+2sm A. stipitatum= 10m+6sm A. haemanthoides= 10m+6sm
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Pharmacognosy and molecular study (ITS marker) of some Allium species (Amaryllidaceae) in Kurdistan province
2021The present study aimed to test pharmacognosy and molecular study of some Allium species (Amaryllidaceae) using internal transcribed spacer (ITS) gene region collected from Saral area of Kurdistan province, Iran. Three Allium species including Allium tripedale, Allium hooshidaryae and Allium stipitatum was selected to be included in the study. Methanolic extracts of selected species was prepared after drying in ppropriate light and temperature conditions. Ferric reducing assay was used to measure antioxidant capacity of extracts. Total phenolic and total flavonoid contents were measured using the Folin Ciocalteau and aluminum chloride color complex methods, respectively. Furthermore, different pharmacogenosy test was performed to predict of present chemical compounds in studied Alliums. Antimicrobial testing of plant extracts for gram-positive and gram-negative strains was performed by disk diffusion method. The results showed a low to moderate antioxidant capacity of studied Alliums compared to ascorbic acid. Among three studied Alliums, the highest antioxidant capacity belonged to Allium hooshidaryae; however, no significant difference was found between three studied Alliums (p>0.05). Pharmacognosy tests indicated that all studied Alliums contained phenolic, flavonoid and glycosides compounds and Allium stipitatum had the highest number of chemical compounds. Antimicrobial tests indicated that Allium stipitatum had greatest inhibitory effects on the bacterial, while Allium hooshidaryae was approximately ineffective. Among the studied bacteria, Pseudomonas aeruginosa strain was approximately resistance against studied Alliums antimicrobial activity. Moreover, Allium stipitatum showed the highest ability to inhibit bacteria growth against Staphylococcus aureus in all concentrations. The results of fragment sequencing revealed that used primers amplified a 900 bps fragment of which 383 nucleotides were vary between used species. The phylogeny analysis showed monophyletic clade for studied species among Melanocrommyum subgenus and close relationship between species. However, they were separated in three different sections.
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بررسی خاصیت آنتی اکسیدانی و ضد میکروبی عصاره گونه هایی از خانواده چتریان در منطقه حفاظت شده کوسالان
2021خانواده چتریان دارای 300 تا 455 جنس و 3000 تا 3750 گونه می باشد. در این تحقیق به مطالعه سه گونه از خانواده چتریان (Zaravschanica membranacea، Anthriscus nemorosa و Zerauchania aucheri) در منطقه حفاظت شده کوسالان پرداخته شده است. گونه های مورد مطالعه در این تحقیق در بهار 1400 از منطقه کوسالان کردستان جمع آوری شدند. در این مطالعه به بررسی خاصیت آنتی اکسیدانی به روش احیای آهن، سنجش میزان فلاونوئید و فنل تام، بررسی فارماگنوزی و آزمایش های ضد میکروبی بر روی گیاهان مذکور پرداخته شده است. ارزیابی توان آنتی اکسیدانی با استفاده از روش احیا آهن نشان داد که گونه Anthriscus nemorosa با داشتن 03/0± 464/0 میلی گرم اسید آسکوربیک در گرم عصاره بیشترین میزان قدرت احیاء کنندگی را داشته است. همچنین این گونه (Anthriscus nemorosa) با داشتن 32/8± 723/112 میلی گرم کوئرستین در گرم عصاره بیشترین مقدار فلاونوئید و با داشتن 35/0± 87/67 میلی گرم اسید گالیک در گرم عصاره بیشترین مقدار فنل تام را داشته است. نتایج بررسی فارماگنوزی نشان می دهد که گونه Zerauchania aucheriدارای بیشترین تعداد متابولیت ثانویه در بین گونه های مورد مطالعه است که شامل ساپونین، فنیک، گلیکوزید و فلاونوئید است. نتایج آزمایش های ضد میکروبی نشان داد که عصاره ی گونه Zerauchania aucheri در غلظت های مختلف برای تکثیر هر سه سویه حساسیتی ایجاد نکرد. بیشترین هاله عدم رشد، مربوط به عصاره گونه Zaravschanica membranacea در غلظت 400 میلی گرم بر روی سویه استافیلوکوکوس اورئوس (حدود 6 میلی متر) بود.
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ارزیابی میکرو آر ان ا های پلاسما به عنوان نشانگر تشخیصی و پیش شناس در سرطان ریه سلول های غیر کوچک در مقایسه با گروه کنترل
2021سرطان ریه یکی از شایع ترین علت مرگ و میر ناشی از سرطان در مردان و زنان در سراسر جهان است. تقریباً 4/1 میلیون مرگ در سال در اثر سرطان ریه اتفاق می افتد. علاوه بر این، میزان مرگ و میر زنان مبتلا به سرطان ریه در کشورهای در حال توسعه به اندازه سرطان دهانه رحم است، که هرکدام 11٪ از کل مرگ و میر ناشی از سرطان زنان را تشکیل می دهند. مطالعات بسیاری با استفاده از بیومارکرها جهت شناسایی اختلافات در بیان miRNA بین بافت های سرطانی و غیر سرطانی مجاور آن، متمرکز بوده است. میکرو RNA ها (miRNA ها) RNA های غیر کد کننده 19 تا 25 نوکلئوتیدی هستند که تنظیم ترجمه بسیاری از ژن ها را کنترل می کنند. miRNA ها در تنظیم فرآیندهای مختلف بیولوژیکی، مانند مرگ سلولی، تکثیر، تمایز سلولی، تنظیم ژن، متابولیسم و تشکیل سرطان نقش دارند. miRNA ها در روند سرطانی شدن سلول ها یا نقش انکوژنیکی دارند (سطح آنها افزایش می یابد) یا نقش سرکوب کننده تومور دارند (سطح آنها کاهش می یابد). در نتیجه می توان از آنها به عنوان نشانگر پیش آگهی و تشخیصی در مراحل اولیه سرطان استفاده کرد. در این مطالعه، بیان ژن های miR144، miR16، miR328، miR200، miR301، miR451، miR340، miR425، miR24، miR126، miR550 و miR378 در نمونه های پلاسمائی بیماران مبتلا به سرطان ریه سلول های غیر کوچک (NSCLC) و نمونه های کنترل با استفاده از Real-time PCR بررسی شده و تغییرات بیان آنها از طریق تکثیر ژن های هدف ارزیابی شده است. نتایج این مطالعه نشان داد که بیان miR144، miR16، miR200، miR301، miR451، miR340، miR425، miR24، miR550 و miR378 در نمونه های تومور نسبت به شاهد کاهش یافته و این miRNA ها به عنوان تومور ساپرسور عمل می کنند. و متعاقباً بیان miR328 و miR126 نمونه های تومور نسبت به شاهد افزایش یافته که این دو miRNA می توانند انکوژن باشند و در مراحل اولیه بیماری، می توان از آنها به عنوان نشانگرهای زیستی برای پیش آگهی و تشخیص سرطانهای ریه سلولهای غیر کوچک استفاده کرد.
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بررسی توالیهای تکراری ریبوزومی نواحی تلومری تعدادی از گونههای جنس لاله واژگون با استفاده از تکنیک هیبریداسیون فلورسنت درجا
2020جنس لاله واژگون ) ( Fritillaria Lمتعلق به تیره سوسنیان ( )Liliaceaاست، که بالغ بر 125گونه با اهمیت دارویی، زینتی و باغبانی را شامل میشود. گونههای این جنس شامل گیاهانی چند ساله، پیازدار و دارای گلهایی واژگون و پرچمهایی بنچسب هستد. لاله واژگون به عنوان یک مدل سیتوژنتیک پیشگام برای مطالعه میوز، میتوز و ساختار کروموزوم، به دلیل داشتن کروموزومهای فوقالعاده بزرگ و همچنین به عنوان جنسی که حاوی بزرگترین ژنوم گیاهی است شناخته شده است. اولین قدم جهت شناخت خصوصیات ژنتیکی یک گیاه تشخیص وضعیت کروموزومی آن گیاه میباشد و به کمک اطلاعات کروموزومی امکان مقایسه گونهها فراهم میگردد. مقایسهی کل DNAکروموزومی هر سلول در بین گونههای مختلف، حاکی از این است که ژنوم یوکاریوتهای عالی حاوی مقادیر زیادی از توالیهای تکرار شونده است، که کدکننده یا غیر کدکننده بوده و در عین حال این توالیها به شدت متغیر هستند. در پژوهش حاضر 6گونه از جنس Fritillariaاز مناطق مختلف استان کردستان جمعآوری شدند و مورد مطالعه کروموزومی و تغیین موقعیت توالیهای تکراری RNAریبوزومی قرار گرفتند. این F. zagrica, F. raddeana, F. persica, F. chlorantha, F. :گونهها شامل که با روش5srDNA و45srDNA هستند. توالیهایavromanica, F. assyriaca کیت-ترجمه نشاندار شده بودند، به عنوان کاوشگر برای تشخیص نواحی سازمان دهنده هستکی در ساختار کروموزوم این گونهها مورد استفاده قرار گرفتند. تکنیک مورد استفاده، هیبریدیزاسیون فلوروسانس درجا ( )FISHبود و پارامترهای کروموزومی و کاریوتایپی به وسیله نرمافزار ایدیوگرام آنالیز شدند. نتایج بدستآمده نشان داد که همه گونهها دیپلویید بوده و عدد کروموزومی آنها 2n=2x=24است. توالیهای هدف در این مطالعه با تنوع در تعداد در نواحی تلومری کروموزومهای گونههای مختلف دیده شدند. کروموزوم Bدر تعداد 4-3عدد، برای نخستین بار در گونه F. assyriacaدر دو جمعیت ( F.assyriacaاکوتیپ آبیدر و اکوتیپ صلواتآباد) گزارش شد و 2عدد Bکروموزوم نیز درگونه F. zagricaمشاهده گردید
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بررسی کروموزومی و کاریوتیپی گونه هایی از جنس Fritillari در ایران
2018جنس Fritillaria L. (لاله واژگون) متعلق به خانواده Liliacea (سوسنیان) است، که شامل 130 گونه پیاز دار و دارای گل های واژگون با خواص دارویی و زینتی است. سیتوژنتیک، دانشی است که اثرات ژنتیکی، ساختار و رفتار کروموزوم ها را مورد مطالعه قرار می دهد. انجام مطالعات سیتوژنتیکی در گیاهان بومی و وحشی اهمیت زیادی دارد، زیرا اولین قدم در جهت شناخت خصوصیات ژنتیکی یک گیاه تشخیص وضعیت کروموزوم های آن گیاه می باشد و به کمک اطلاعات کروموزومی امکان مقایسه گونه ها و جمعیتها فراهم می گردد. در پژوهش حاضر گونه های متفاوت از جنس Fritillaria در مناطق مختلف از استان کردستان مورد مطالعه قرار گرفت. گونه های جمع آوری شده شامل: F. avramanica، F. assyriaca، F. imperialis، F. persica، F. chlorantha، F. raddeana می باشند. در این تحقیق ساختار کروموزوم های گونه ها با استفاده از روش های سیتوژنتیک کلاسیک مثل اسکواشینگ و سیتوژنتیک مولکولی مثل هیبریداسیون فلوروسنت درمحل (فیش) مورد مطالعه قرار گرفتند. برای برآورد پارامترهای کروموزومی و کاریوتیپی از نرم افزار ایدوکار استفاده شد. حداقل 5 پلاک سلول متافازی از هرگونه مورد تجزیه و تحلیل قرار گرفتند. برای انجام روش فیش از پلاسمید pTA71 که توالی های مربوط به ژن های S rDNA45 ریبوزومی گندم در آن کلون شده بود، به عنوان کاوشگر برای تشخیص نواحی سازمان دهنده هستکی استفاده شد. نتایج فیش برای دو گونه F. persica و F. raddeana نواحی سازمان دهنده هستکی را نشان داد. نتایج نشان داد که همه گونه ها دیپلوئید بود و عدد پایه کروموزومی 12 x= است. در بیشتر گونه ها کروموزوم ماهواره دار مشاهده شد. به طور عمده ماهواره آنها بر روی بازوی بلند کروموزوم است. در این مطالعه تصویر میکروسکوپی پلاک های کروموزومی متافازی، ایدیوگرام و مشخصات کاریوتیپ گونه های مورد مطالعه ارائه شد. لازم به ذکر است گونه F.avramanica برای اولین بار در دنیا ازنظر سیتوژنتیکی مورد مطالعه قرارگرفته است. فرمول کاریوتیپی گونه ها به شرح ذیل تعیین گردید: F. avramanica = 2sm+ 20St+ 2t F. assyriaca = 8sm + 14st+ 2t F. raddeana = 4m+2sm+4st+14t F. persica = 4m+2sm+16st+2t F. chlorantha = 6m+6sm+12t