دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 Genetic Diversity of Hesperis persica, Based On Inter-Retrotransposon Amplified Polymorphism تنوع ژنتیکی Hesperis persica ، بر اساس DNA چند شکلی تقویت شده بین رتروترانسپوزونی (IRAP) 322 336 731556 10.66224/JCT.16.4.322 FA سمیه اسفندانی بزچلوئی پژوهشکده فناوری نوین زیستی، دانشگاه زنجان، زنجان، ایران Journal Article 2025 02 05 Introduction: Knowledge of spatial genetic structures provides a valuable tool for inferring the evolutionary forces such as selective pressures and drift. Low gene flow due to spatial isolation of populations may even increase the degree of local differentiation. Nevertheless, phenotypic plasticity rather than genetic differentiation may be an alternative way of matching genotypes to environment; indeed increasing environmental variation favors higher levels of plasticity.Genetic diversity is one aspect of biological diversity that is extremely important for conservation strategies, especially in rare and narrowly endemic species. The genus Hesperis L. (Brassicaceae) comprises biennial and perennial herbs and consists of 46 species worldwide, mainly occurring in different parts of Europe, Caucasus, Transcaucasia, and to a lesser extent in northern and central Asia, and mostly in Turkey with 28 species. The genus is represented by 11 or six species belonging to sections Hesperis Dvořák, Diaplictos Dvořák and Pachycarpos Fourn. in Iran. Aim: Moreover, due to extensive morphological variability of this species in the country, there is possibility of having infra-specific taxonomic forms in this species. Therefore, we carried out population genetic analysis and morphometric study of 11 geographical populations for the first time in the country. For genetic study, we used the inter-retrotransposon amplified polymorphism (IRAP) method that displays insertional polymorphisms by amplifying the segments of DNA between two retrotransposons. It has been used in numerous studies of genetic diversity. The objectives of this research were to study genetic diversity Hesperis persica with a different geographical origin by inter-retrotransposon amplified polymorphism (IRAP) method. Materials and Methods: A total of 73 individuals were sampled representing 11 natural populations of H. persica Boiss. subsp. persica and H. persica subsp. kurdica (F. Dvořák & Hadac) F. Dvořák, in Mazandaran, East Azerbaijan, Kohgilouye-Boirahmad, Chaharmahal Bakhtiari, Fars, Zanjan, Tehran, Kordestan, Provinces of Iran during July-Agust 2019-2024. Fresh leaves were used randomly from 6-10 plants in each of the studied populations. These were dried by silica gel powder. CTAB activated charcoal protocol was used to extract genomic DNA. The quality of extracted DNA was examined by running on 0.8% agarose gel. A set of six outward-facing LTR primers were used for IRAP analysis. We also used 15 different combinations of outward-facing LTR pair primers. PCR reactions were carried in a 25μl volume containing 10 mM Tris-HCl buffer at pH 8; 50 mM KCl; 1.5 mM MgCl2; 0.2 mM of each dNTP (Bioron, Germany); 0.2 μM of a single primer; 20 ng genomic DNA and 3 U of Taq DNA polymerase (Bioron, Germany). The thermal program was carried out with an initial denaturation for 1 min at 94°C, followed by 40 cycles in three segments: 35 s at 95°C, 40s at 47°C and 55s at 72°C. Final extension was performed at 72°C for 5 min. The amplification products were observed by running on 1% agarose gel, followed by the ethidium bromide staining. The fragment size was estimated by using a 100 bp molecular size ladder (Fermentas, Germany).The IRAP profiles obtained for each samples were scored as binary characters. Parameter like Nei’s gene diversity (H), Shannon information index (I), number of effective alleles, and percentage of polymorphism were determined.Nei’s genetic distance among populations was used for Neighbor Joining (NJ) clustering and Neighbor-Net networking. Mantel test checked the correlation between geographical and genetic distance of the studied populations . These analyses were done by PAST ver. 2.17 , DARwin ver. 5 (2012) and SplitsTree4 V4.13.1 (2013) software. AMOVA (Analysis of molecular variance) test (with 1000 permutations) as implemented in GenAlex 6.4, and Nei,s Gst analysis as implemented in GenoDive ver.2 (2013) were used to show genetic difference of the populations. Moreover, populations, genetic differentiation was studied by G'ST est = standardized measure of genetic differentiation, and D_est = Jost measure of differentiation. The genetic structure of populations was studied by Bayesian based model STRUCTURE analysis , and maximum likelihood-based method of K-Means clustering of GenoDive ver. 2. (2013). For STRUCTURE analysis, data were scored as dominant markers. The Evanno test was performed on STRUCTURE result to determine proper number of K by using delta K value. In K-Means clustering, two summary statistics, pseudo-F, and Bayesian Information Criterion (BIC), provide the best fit for k. Results: The highest value of percentage polymorphism (57.41%) was observed in Mazandaran, 30 km S. of Ramsar between Kash-e Chal mountain and Miankuh (population No.8, H. persica subsp. kurdica) which shows high value for gene diversity (0.34) and Shanon, information index (0.43). Population Chaharmahal Bakhtiari, Shahr-e Kurd, tang-e Sayyad protected area, Pir kuh mountain (No.3, H. persica subsp. persica) has the lowest value for percentage of polymorphism (28.11%) and the lowest value for Shanon, information index (0.088), and He (0.022).AMOVA (PhiPT = 0.98, P = 0.010), and Gst analysis (0.654, p = 0.001) revealed significant difference among the studied populations. It also revealed that, 40% of total genetic variability was due to within population diversity and 60% was due to among population genetic differentiation. Pairwise AMOVA produced significant difference among the studied populations. Moreover, we got high values for Hedrick standardized fixation index after 999 permutation (G’st = 0.654, P = 0.001) and Jost, differentiation index (D-est = 0.769, P = 0.001). These results indicate that the geographical populations of Hesperis persica are genetically differentiated from each other. The mean Nm = 0.455 was obtained for all IRAP loci, which indicates low amount of gene flow among the populations. Population assignment test also agreed with Nm result and could not identify significant gene flow among these populations. However, reticulogram obtained based on the least square method, revealed some amount of shared alleles among populations 2 and 3, and between 7 and 8, also between 1, and 4. This result is in agreement with grouping we obtained with PCoA plot, as these populations were placed close to each other. As evidenced by STRUCTURE plot based on admixture model, these shared alleles comprise very limited part of the genomes in these populations and all these results are in agreement in showing high degree of genetic stratification within Hesperis persica populations. In total 76 IRAP bands (loci) were obtained, out of which 14 bands were private. Populations 2 and 5-7 contained 2-5 private bands. Conclusion: PCoA plot of populations was in agreement with WARD clustering of molecular data. These results indicated that geographical populations of Hesperis persica are well differentiated based on (IRAP) markers. هدف : تنوع ژنتیکی یکی از جنبه‌های تنوع زیستی است که برای استراتژی‌های حفاظتی، به‌ویژه در گونه‌های کمیاب و اندمیک بسیار مهم است . جنس Hesperis L. (Brassicaceae) شامل گیاهان دو ساله و چند ساله است و از 46 گونه در سراسر جهان تشکیل شده است که عمدتاً در مناطق مختلف اروپا، قفقاز، ماوراء قفقاز و به میزان کمتر در شمال و آسیای مرکزی و به میزان بیشتر در ترکیه با 28 گونه یافت میشود. این جنس با 11 یا 6 گونه متعلق به بخش های Hesperis Dvořák, Diaplictos Dvořák and Pachycarpos Fourn. نشان داده می شود. مواد و روش ها: با توجه به اهمیت این گونه، در مجموع 73 فرد به نمایندگی از 11 جمعیت طبیعی H. persica Boiss. subsp. persica and H. persica subsp. kurdica (F. Dvořák & Hadac) F. Dvořák, نمونه برداری شدند. برای شناسایی صحیح گونه (Hesperis persica) از مراجع مختلفی استفاده شد. نتایج: آزمون AMOVA تفاوت ژنتیکی معنی‌داری (88/0=PhiPT، 010/0=P) در بین جمعیت‌های مورد مطالعه ایجاد کرد و همچنین نشان داد که 40 درصد از کل تنوع ژنتیکی به دلیل تنوع درون جمعیتی و 60 درصد به دلیل تمایز ژنتیکی بین جمعیت است. نتیجه‌گیری: نمودار PCoA جمعیت ها با خوشه بندی WARD داده های مولکولی مطابقت داشت. این نتایج نشان داد که جمعیت های جغرافیایی Hesperis persica بر اساس نشانگرهای (IRAP) به خوبی متمایز می شوند.

https://jct.araku.ac.ir/article_731556_03f8ba297808ac032311548cfe7fdf3b.pdf

دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 The effect of Paxlovid on skeletal system morphogenesis in a rat animal model (histological study) اثر داروی پاکسلوید بر مورفوژنز سیستم اسکلتی در مدل حیوانی رت ( مطالعه بافت‌شناسی) 337 351 730599 10.66224/JCT.16.4.337 FA علی عبداللهی دانشجوی کارشناسی ارشد زیست شناسی سلولی و تکوینی، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه اردکان، اردکان، ایران 0009-0005-9545-0649 مجید مروتی-شریف‌آباد دانشیار، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه اردکان، اردکان، ایران 0000-0002-4568-1562 الهام صالحی دانشیار، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه اردکان، اردکان، ایران 0000-0003-3004-1252 محسن رشیدی دکتری گروه بافت شناسی، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران 0009-0008-6253-6309 علی رضایی گلمیشه استادیار، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه اردکان، اردکان، ایران حسن مروتی استاد گروه بافت شناسی، گروه علوم پایه، دانشکده دامپزشکی، دانشگاه تهران، تهران، ایران 0000-0003-0275-1636 Journal Article 2025 05 05 Introduction: Paxlovid, an investigational oral therapeutic combining the antiviral Nirmatrelvir and the pharmacokinetic enhancer Ritonavir, is being developed to treat SARS-CoV-2 infection, aiming to reduce severe disease progression, hospitalization, and mortality. While its efficacy has been shown in Phase 2/3 trials for non-hospitalized patients, its safety, particularly regarding fetal development, remains under investigation. Coronaviruses have long posed significant challenges to global public health, with their potential to cause severe respiratory infections in humans. Over the past two decades, two novel coronaviruses—Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV)—have emerged, leading to widespread morbidity and mortality. The SARS - CoV outbreak, which occurred in the early 2000s, infected over 8,000 individuals worldwide and resulted in nearly 800 deaths, representing a mortality rate of approximately 10 %. Similarly, MERS - CoV, identified in 2012, caused 857 confirmed infections and 3 34 deaths, with a strikingly high mortality rate of around 35 %. These outbreaks underscored the persistent threat posed by coronaviruses to human health. Aims: Pregnant women represent a particularly vulnerable population during infectious disease outbreaks, and the potential teratogenic effects of new therapeutics must be carefully evaluated. Previous studies on Nirmatrelvir have explored its effects on skeletal morphogenesis, but comprehensive data on Paxlovid's impact on fetal development remain limited. This study aims to address this gap by investigating the effects of Paxlovid on the reproduction and skeletal morphogenesis of rat embryos, providing critical insights into its safety profile during pregnancy. By elucidating the potential risks associated with Paxlovid, this research contributes to the broader effort to ensure the safe use of antiviral therapies in vulnerable populations. This study assessed the potential embryotoxic effects of Paxlovid on rat fetal skeletal morphogenesis, following ICH guidelines. Materials and methods: Pregnant rats were divided into four groups, receiving doses of 0, 100/60, 300/200, or 1000 mg/kg/day of Paxlovid. Maternal clinical symptoms and weight were monitored, and fetal outcomes, including weight, crown-rump length (CRL), and abdominal circumference (AC), were evaluated on gestational day 21. Histological analysis of fetal skeletal tissue was conducted using hematoxylin-eosin and Alizarin Red S staining to detect structural abnormalities. Results: Results showed reduced maternal weight gain in Paxlovid-treated groups compared to controls. Fetuses in treated groups also exhibited lower weight, CRL, and AC. However, histological analysis revealed no structural abnormalities in skeletal tissue. These findings suggest that while Paxlovid may transiently affect fetal growth metrics, it does not induce teratogenic effects on skeletal development. Discussion: The findings of this study revealed no histological abnormalities in the skeletal system of fetuses exposed to Paxlovid. The observed effects were limited to reductions in fetal weight, crown-rump length (CRL), and abdominal circumference (AC). Collectively, the data from this investigation, along with existing evidence, suggest a low risk of fetal harm associated with Paxlovid, which comprises Nirmatrelvir (NMV), a potent and selective inhibitor of the SARS-CoV-2 main protease, and Ritonavir, a pharmacokinetic enhancer. Nonetheless, the use of Paxlovid during pregnancy should be carefully evaluated, with a thorough discussion of potential risks and benefits conducted in consultation with a healthcare professional. Conclusion: The study concludes that Paxlovid does not compromise the histological integrity of fetal skeletal tissue, supporting its potential safety during pregnancy. However, further research is needed to understand the mechanisms behind the observed growth effects and to confirm these findings in human populations. Overall, the results indicate a low risk of fetal harm, reinforcing Paxlovid's safety profile for use in pregnant individuals هدف: پاکسلووید، یک داروی ترکیبی خوراکی حاوی نیرماترلویر (عامل ضد ویروسی) و ریتوناویر (تقویت کننده فارماکوکینتیک)، برای درمان کووید-۱۹ با هدف کاهش شدت بیماری، بستری و مرگ و میر توسعه یافته است. با این‫حال، ایمنی آن در دوران بارداری، به‫ویژه تاثیر آن بر رشد جنین، نیاز به بررسی دارد. این مطالعه با رعایت دستورالعمل‌های ICH، اثرات احتمالی پاکسلووید بر مورفوژنز اسکلتی جنین موش را ارزیابی کرد. مواد و روش‏ها: رت‌های باردار به چهار گروه تقسیم شدند. گروه اول(کنترل) دارویی دریافت نکرد، گروه دوم دوزهای 60 و 100، گروه سوم دوزهای 200 و 300 و گروه چهارم دوز ثابت 1000 میلی‌گرم بر کیلوگرم روزانه پاکسلووید را دریافت کردند. پارامترهای مادرانه (وزن و علائم بالینی) و جنینی( وزن، طول سر-دم (CRL) و دور شکم (AC)) در روز ۲۱ بارداری بررسی شدند. ناهنجاری‌های اسکلتی با رنگ‌آمیزی هماتوکسیلین-ائوزین و آلیزارین رد S ارزیابی شدند. نتایج: کاهش وزن مادران و جنین‌ها در گروه‌های تحت درمان نسبت به گروه کنترل مشاهده شد، اما آنالیز بافت‌شناسی اسکلت، ناهنجاری ساختاری نشان نداد. این یافته‌ها حاکی از آن است که پاکسلووید اگرچه ممکن است بر شاخص‌های رشد جنین تاثیر گذارد، اما اثر تراتوژنیک بر اسکلت ندارد. نتیجه‏گیری: مطالعه حاضر نشان می‌دهد که پاکسلووید یکپارچگی اسکلت جنین را مختل نمی‌کند و از ایمنی نسبی آن در بارداری حمایت می‌کند. با این حال، پژوهش‌های بیشتر برای درک مکانیسم‌های تأثیر بر رشد و تعمیم نتایج به انسان ضروری است. به‌طورکلی، یافته‌ها حاکی از خطر پایین آسیب جنینی بوده و ایمنی پاکسلووید را در جمعیت‌های باردار تقویت می‌کنند.

https://jct.araku.ac.ir/article_730599_b21a0dcfda76ae02e85681f53f0b6706.pdf

دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 The Synergistic Effect of Stem Cell-Derived Exosomes and Glucosamine on the Expression of Sox9, Acan, Col2a1, and Col10a1 Genes in Bone Marrow Mesenchymal Stem Cells of NMRI Mice in a Chondrogenic Medium اثر هم افزایی اگزوزوم‫های مشتق از سلول‫های بنیادی و گلوکزآمین بر بیان ژن‫های Sox9، Acan، Col2a1 و Col10a1 در سلول‫های بنیادی مزانشیمی مغز استخوان موش نژاد NMRI در محیط کندروژنیک 352 368 733300 10.66224/JCT.16.4.352 FA مریم لطفی گروه زیست شناسی، دانشکده علوم پایه، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران،maryam.lotfi0043@iau.ac.ir 0009-0003-1443-6067 جواد بهارآرا گروه زیست شناسی و مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران، baharara@iau.ac.ir خدیجه نژاد شاهرخ آبادی گروه زیست شناسی، دانشکده علوم پایه، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران، Shahrokhabady@yahoo.com 0000-0002-3536-3456 پژمان خورشید گروه فیزیک، دانشکده علوم پایه، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران، Pejman.khorshid@iau.ac.ir 0000-0003-4342-2759 Journal Article Introduction: Cartilage, a tissue without blood vessels and nerves, possesses inherently limited regenerative capacity following injury, often leading to progressive joint degeneration and conditions like osteoarthritis (OA) if left untreated. Current clinical interventions, such as surgical microfracture or autologous chondrocyte implantation (ACI), face significant challenges, including donor site morbidity, immune rejection, and the formation of fibrocartilage with inferior biomechanical properties. These limitations underscore the urgent need for novel therapeutic strategies that can effectively stimulate hyaline cartilage regeneration. In this context, mesenchymal stem cell-derived exosomes (MSC-Exos) have garnered attention as a cell-free regenerative tool, leveraging their cargo of bioactive molecules (e.g., miRNAs, cytokines, and growth factors) to modulate chondrogenesis, suppress inflammation, and enhance extracellular matrix (ECM) synthesis. Concurrently, glucosamine, a natural amino sugar and precursor for glycosaminoglycan (GAG) biosynthesis, has demonstrated dual functionality in joint health: not only does it serve as a building block for proteoglycans critical to cartilage integrity, but it also exhibits chondroprotective effects by mitigating ECM degradation and promoting stem cell chondrogenic differentiation. The potential synergy between MSC-Exos and glucosamine could thus address multiple facets of cartilage repair, combining anabolic stimulation (via exosomal signaling) with metabolic support (via glucosamine supplementation), offering a promising combinatorial approach to halt OA progression and restore functional cartilage. Aims: This study aimed to investigate the combined effect of mouse bone marrow stem cell-derived exosomes and glucosamine on the expression of cartilage-specific genes, including Sox9, Acan, Col2a1, and Col10a1. Materials and Methods: Bone marrow mesenchymal stem cells were prepared from NMRI mice. The mice were euthanized by cervical dislocation, the femoral heads were removed, and the bone marrow contents were transferred into a cell culture flask using a syringe containing culture medium. The bone marrow cells were cultured and were ready for use after 3 to 5 passages. The cell supernatant was separated, and exosomes were extracted from it by successive rounds of centrifugation followed by ultracentrifugation. Mesenchymal stem cell viability and determining the appropriate concentration of exosomes and glucosamine were performed using the MTT assay. The experiments were performed on mesenchymal stem cells in 4 groups: control, exosome, glucosamine, and exosome + glucosamine. The effects of exosomes and glucosamine on the expression of Sox9, Acan, Col2a1, and Col10a1 genes in mesenchymal stem cells were investigated in the presence of chondrogenic medium. Results: According to the MTT assay results demonstrating the synergistic effect of exosomes and glucosamine, the combined concentrations of 15 μg/mL exosomes and 25 μg/mL glucosamine were chosen for subsequent applications. Real-time PCR results showed that the expression of Sox9, Acan, and Col2a1 genes in stem cells treated with exosomes and glucosamine significantly increased compared to the other groups after 14 days, while the expression of the Col10a1 gene significantly decreased compared to the other groups. Discussion: The combined treatment of bone marrow–derived mesenchymal stem cell (BMSC) exosomes and glucosamine significantly upregulated the expression of key chondrogenic markers, including Sox9, Acan, and Col2a1, while downregulating the hypertrophic marker Col10a1. This gene expression profile suggests a dual beneficial effect: (1) promotion of chondrogenic differentiation and extracellular matrix (ECM) synthesis, and (2) suppression of hypertrophic differentiation, a critical factor in preventing cartilage calcification and osteoarthritis progression. These findings highlight the synergistic potential of BMSC exosomes and glucosamine as a combinatorial therapy for cartilage regeneration. By enhancing anabolic processes (Sox9-mediated chondrogenesis and aggrecan/collagen II deposition) and concurrently inhibiting catabolic pathways (Col10a1-associated hypertrophy), this strategy may offer a promising approach to delay or reverse early-stage cartilage degeneration in degenerative joint diseases Conclusion: Our study reveals that combining bone marrow stem cell-derived exosomes with glucosamine synergistically enhances chondrogenesis by upregulating key cartilage markers (Sox9, Acan, Col2a1) while suppressing hypertrophy-related Col10a1. This dual action suggests that exosomes promote cartilage matrix synthesis through their bioactive cargo (e.g., miRNAs/growth factors), while glucosamine likely inhibits hypertrophic differentiation, potentially via modulation of the Wnt/β-catenin pathway. These findings support this combination as a promising strategy for improving cartilage repair and preventing OA progression, though further in vivo validation is needed. هدف: غضروف‫ها فاقد رگ خونی و دارای توانایی ترمیم محدود می‫باشند. بازسازی آسیب غضروفی به‫عنوان یک چالش باقی مانده است. اگزوزوم‫ها وزیکول‫های خارج سلولی می‫باشند که از انواع مختلفی از سلول‫ها آزاد می‫شوند. از طرف دیگر مشاهده شده که گلوکزآمین به‫عنوان دارو نقش مهمی در بازسازی غضروف ایفا می‫کند. هدف از این پژوهش بررسی تاثیر توام اگزوزوم‫های مشتق از سلول‫های بنیادی مغز استخوان موش و گلوکز آمین بر بیان ژن‫های اختصاصی غضروف می‫باشد. مواد و روش‏ها: سلول‫های بنیادی مزانشیمی مغز استخوان موش کوچک آزمایشگاهی کشت داده شدند و اگزوزوم‫ها از محیط رویی سلول‫ها به‫روش التراسانتریفیوژ استخراج شد. اثرات اگزوزوم و گلوکزآمین بر بیان ژن‫های Sox9، Acan، Col2a1 و Col10a1 در سلول‫های بنیادی مزانشیمی در حضور محیط کندروژنیک بررسی شد. نتایج: نتایج شناسایی، حضور اگزوزوم‫ها با اندازه‫ی تقریبی 100 نانومتر را تایید کردند و همچنین نتایج Real time PCR نشان داد که بیان ژن‫های Sox9، Acan و Col2a1 در سلول‫های بنیادی تحت تیمار توام با اگزوزوم و گلوکزآمین نسبت به سایر گروه‫ها پس از 14 روز افزایش معنی دار و همچنین بیان ژن Col10a1 کاهش معنادار داشته است. نتیجه‏گیری: ترکیب اگزوزوم‫‌های مشتق از سلول‌های بنیادی مغز استخوان و گلوکوزامین باعث افزایش بیان ژن‌های اختصاصی غضروف (Sox9، Acan و Col2a1) و کاهش بیان ژن هیپرتروفیک Col10a1 شد که نشان‌دهنده تقویت فرایند کندروژنز و مهار تمایز هیپرتروفیک است. این نتایج نشان می‌دهد که استفاده هم‫زمان از این دو عامل می‌تواند رویکردی موثر برای بازسازی غضروف و پیشگیری از تخریب زودرس آن باشد.

https://jct.araku.ac.ir/article_733300_de9ed3e83dda8ed7ae0bb0c6f772c64e.pdf

دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 Curcumin Reinstates Osteogenic Differentiation Disrupted by DEHP in Rat Mesenchymal Stem Cells کورکومین تمایز استئوژنیک مختل‌شده توسط DEHP را در سلول‌های بنیادی مزانشیم رت بازسازی می‌کند 369 388 730596 10.66224/JCT.16.4.369 FA محمدحسین آبنوسی دانشگاه اراک، دانشکده علوم پایه، گروه زیست شناسی 0000-0001-6655-1890 محجوبه لک گروه زیست شناسی، دانشکده علوم پایه، دانشگاه اراک، اراک، ایران 0009-0000-9158-8687 Journal Article 2025 07 21 Introduction: Di(2-ethylhexyl) phthalate (DEHP) is a commonly used plasticizer in polyvinyl chloride (PVC) products, including medical devices such as syringes, IV tubing, blood bags, respiratory circuits, and dialysis equipment. Due to its weak physical bonding to the PVC matrix and lack of covalent interactions, DEHP can leach into biological fluids upon contact. This becomes particularly concerning during prolonged clinical exposure, as it allows the compound to enter the bloodstream and potentially affect sensitive tissues, including bone marrow. Bone marrow-derived mesenchymal stem cells (BMSCs) are multipotent progenitors with the ability to differentiate into osteoblasts, playing a vital role in bone remodeling and regeneration. Given their sensitivity to environmental toxins, DEHP exposure represents a significant risk to BMSC viability and osteogenic function. Evidence suggests that DEHP disrupts osteogenesis by inducing oxidative stress and downregulating essential genes involved in matrix formation and mineralization. Curcumin (Cur), a bioactive polyphenol extracted from the rhizome of Curcuma longa, possesses strong antioxidant, anti-inflammatory, and cytoprotective properties. It is nontoxic, affordable, and widely available, making it a promising candidate for counteracting xenobiotic-induced oxidative damage. This study explores whether Curcumin can mitigate DEHP’s deleterious effects on BMSCs during osteogenic differentiation. Aim: We hypothesize that co-treatment with Cur will ameliorate DEHP-mediated disruption in cell viability, antioxidant capacity, and osteogenic marker expression. Materials and Methods: BMSCs were isolated from adult male Wistar rats under sterile conditions and cultured to passage three. Cells were divided into four experimental groups and exposed for 21 days to the following treatments: (1) control, (2) DEHP (100 μM), (3) Curcumin (0.1 μM), and (4) DEHP + Curcumin. The cells were cultured in osteogenic differentiation medium throughout the treatment period. Cell viability was assessed using the tetrazolium-based MTT assay. Osteogenic differentiation was evaluated via Alizarin Red staining for mineral deposition, calcium quantification, and alkaline phosphatase (ALP) activity measurement. Oxidative stress was assessed by quantifying intracellular malondialdehyde (MDA) levels, a lipid peroxidation marker. Antioxidant enzyme activity for catalase (CAT) and superoxide dismutase (SOD) was measured spectrophotometrically. Total antioxidant capacity (TAC) was evaluated using commercial kits. For molecular analysis, total RNA was extracted from cells and reverse-transcribed into complementary DNA (cDNA). Semi-quantitative PCR was performed to measure expression levels of osteogenic differentiation-related genes: Smad1, Bmp2, Bmp7, Runx2, Alp, Col-1A1, and Osteocalcin (Oc). Gapdh served as the internal control. Data were statistically analyzed using ANOVA with Tukey’s post-hoc test. A threshold of p < 0.05 was considered statistically significant. Results DEHP treatment caused a marked reduction in BMSC viability (p < 0.0001), confirming its cytotoxicity. Cells co-treated with Curcumin showed a significant restoration in viability (p < 0.01), indicating Cur’s protective effects. Alizarin Red staining revealed diminished extracellular matrix mineralization in DEHP-treated cells, with corresponding reductions in calcium content and ALP activity. Notably, Curcumin co-treatment restored all markers to levels comparable to those seen in control cells. ALP, an early osteogenic marker, showed complete recovery, underlining Cur’s efficacy in preserving bone-forming potential. Gene expression analysis demonstrated that DEHP downregulated osteogenic genes (Smad1, Bmp2, Bmp7, Runx2, Col-1A1, Oc). Co-treatment with Curcumin significantly reversed this suppression, elevating transcript levels to near control values. These results suggest a transcriptional rescue linked to improved redox homeostasis. Oxidative stress measurements indicated that DEHP increased MDA levels while suppressing CAT and SOD activity, as well as total antioxidant capacity (p < 0.0001). Curcumin treatment effectively reduced MDA concentrations (p < 0.05), and boosted CAT and SOD activity (p < 0.01). TAC was significantly elevated in Curcumin-treated groups (p < 0.0001), indicating improved redox balance and defense against oxidative damage. Conclusion: This study provides strong evidence that DEHP impairs BMSC viability and osteogenic differentiation primarily through oxidative stress mechanisms. Curcumin co-treatment mitigates DEHP-induced cellular damage, restores osteogenic function, and enhances antioxidant defenses. These findings highlight Curcumin’s potential as a therapeutic agent to counteract phthalate toxicity in clinical contexts involving prolonged exposure to DEHP-containing materials. هدف: دی-2-اتیلهگزیل فتالات (DEHP) به‌عنوان نرم‌کننده در محصولات PVC پزشکی استفاده می‌شود و به‌دلیل عدم اتصال کووالانسی، در فرایند درمان جدا شده و موجب آلودگی مغز استخوان می‌شود. مطالعات نشان داده‌اند که DEHP با القای استرس اکسیداتیو، تمایز استئوژنیک سلول‌های بنیادی مزانشیمی (BMSCs) را مهار می‌کند. در این پژوهش، نقش آنتی‌اکسیدانتی کورکومین (Cur) در کاهش آسیب‌های DEHP بررسی شده است. مواد و روش‌ها: سلول‌ها BMSC از رت ویستار استخراج و پس از پاساژ سوم، به‌مدت ۲۱ روز در محیط استئوژنیک با DEHP (100 μM)، کورکومین (0.1 μM) و ترکیب هم‫زمان آن‫ها تیمار شدند. توانائی زیستی و تمایز استئوژنیک با آزمون‌های آلیزارین رد، کلسیم ماتریکس و آلکالین فسفاتاز ارزیابی شد. استرس اکسیداتیو با اندازه‌گیری MDA، فعالیت CAT و SOD و سطح TAC بررسی گردید. همچنین بیان ژن‌های استئوژنیک از طریق RT-PCR برای ژن‌های SMAD1، BMP2، BMP7، RUNX2، COL-1، ALP، OC و GAPDH انجام شد. نتایج: DEHP موجب کاهش شدید توانائی زیستی و تمایز استئوژنیک BMSCs شد (p<0.0001)، در حالی‫که کورکومین در تیمار هم‫زمان این اثرات را به‌طور معنی‌دار جبران نمود، به‌طوری‌که ماتریکس استخوانی، غلظت کلسیم و فعالیت آلکالین فسفاتاز به سطح کنترل رسید. همچنین بیان همه ژن‌های دخیل در تمایز استئوژنیک افزایش یافت و Cur موجب کاهش MDA (p<0.05) و افزایش فعالیت CAT، SOD و TAC شد، و توانست اثرات استرس اکسیداتیو ناشی از DEHP را خنثی کند. نتیجه‌گیری: کورکومین با مهار استرس اکسیداتیو توانست اثرات مهاری DEHP بر تمایز استئوژنیک BMSCs را جبران نماید. لذا پیشنهاد می‌شود درصورت مواجهه با DEHP در طی درمان دراز مدت از کورکومین استفاده شود.

https://jct.araku.ac.ir/article_730596_1aa57dc96cf7389f98a92b839e1fbb39.pdf

دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 Strategies to deal with heat stress in crop plants راه‫کارهای مقابله با تنش گرما در گیاهان زراعی 389 420 733406 10.66224/JCT.16.4.389 FA اسماعیل قلی نژاد دانشیار گروه علمی علوم کشاورزی، دانشگاه پیام نور، تهران، ایرانe_gholinejad@pnu.ac.ir 0000-0003-1896-8784 رضا درویش زاده استاد، گروه تولید و ژنتیک گیاهی دانشکده کشاورزی دانشگاه ارومیه. ارومیه: r.darvishzadeh@urmia.ac.ir Journal Article Introduction: Heat stress is one of the most significant environmental stresses that limits the growth, metabolism, and productivity of crops worldwide. As global temperatures rise due to climate change, the intensity and frequency of hot and dry days are increasing significantly. This phenomenon poses a serious threat to agricultural productivity, as the simultaneous occurrence of drought and heat stress adversely affects various agricultural characteristics. These include traits related to growth and development, biomass accumulation, and overall yield. In this context, various physiological traits such as leaf water content, canopy temperature, membrane stability, chlorophyll content, stomatal conductance, chlorophyll fluorescence, and photosynthesis are seriously disrupted. Understanding these impacts is crucial for developing effective strategies to mitigate heat stress and enhance crop resilience. The objective of this article is to investigate the effects, mechanisms of tolerance, management, and control of heat stress in crop plants. This article is prepared as a review of the literature and examines various strategies for coping with heat stress in plants. This article is a review article that was obtained by searching related articles in reliable sites (Google Scholar, Web of Science, PubMed, Scopus, and SID) and aims to investigate the effects, mechanisms of tolerance, management, and control of heat stress. Plants have developed a range of adaptive defense strategies to cope with heat stress. These strategies include mechanisms for removing reactive oxygen species (ROS), producing osmolytes, and modulating secondary metabolites and various hormones. The survival of the plant under heat stress depends on its ability to perceive the stress, produce and transmit signals, and initiate appropriate physiological and biochemical changes. For instance, changes in gene expression and metabolite synthesis significantly improve plant tolerance to heat stress. Adaptation mechanisms to heat stress include leaf curling, which reduces water loss, precocity, which allows for earlier maturation, and the accumulation of osmotic protectors that help maintain cellular integrity. Additionally, the activation of antioxidant defense mechanisms plays a crucial role in mitigating oxidative damage caused by heat stress. Heat stress can be effectively mitigated through various agricultural practices. These practices include selecting appropriate planting methods, choosing the right planting date, selecting suitable cultivars that are more resilient to heat, and implementing effective irrigation methods. Furthermore, the exogenous use of protectants, such as osmotic protectors (e.g., proline, glycine betaine, trehalose), phytohormones (e.g., abscisic acid, gibberellic acids, jasmonic acids), signaling molecules (e.g., nitric oxide), polyamines (e.g., putrescine, spermidine, spermine), trace elements (e.g., selenium, silicon), and essential nutrients (e.g., nitrogen, phosphorus, potassium, calcium) are effective in reducing the damage caused by heat stress. These practices not only enhance plant resilience but also contribute to maintaining agricultural productivity under changing climatic conditions. Conclusion: Molecular and biotechnological strategies are also crucial for developing heat stress tolerance in plants. Advances in molecular biology have facilitated a better understanding of the mechanisms underlying heat stress tolerance. Plants respond to environmental stresses by modulating the expression of multiple genes and coordinating gene expression in various ways. The expression of heat shock proteins (HSPs) plays a vital role in protecting intracellular proteins from denaturation, thereby maintaining their stability and function. By integrating molecular approaches with traditional breeding techniques, researchers can develop crop varieties that are better equipped to withstand heat stress. Overall, a comprehensive understanding of heat stress mechanisms and effective management strategies is essential for ensuring sustainable agricultural productivity in the face of climate change. مقدمه: تنش گرمایی، با تشدید ناشی از تغییرات اقلیمی، یکی از مهم‌ترین محدودیت‌های رشد و بهره‌وری گیاهان است. وقوع همزمان گرما و خشکی تأثیرات مضاعفی بر ویژگی‌های مورفولوژیک، فیزیولوژیک و عملکرد محصولات زراعی دارد. مقاله حاضر یک مقاله مروری است که با جستجو در مقالههای مرتبط در سایتهای معتبر (Google scholar, Web of science, PubMed, Scopus, sid) به‫دست آمده است و با هدف بررسی اثرات، سازوکارهای تحمل، مدیریت و کنترل تنش گرما تهیه شده است.گیاهان برای مقابله با تنش گرمایی از راهبردهای اجتناب و سازگاری استفاده می‌کنند. اجتناب شامل واکنش‌های سریع مانند تنظیم روزنه‌ها، تغییر جهت برگ‌ها و خنک‌سازی از طریق تعریق است. سازگاری‌های بلندمدت نیز تغییرات تکاملی در صفات فنولوژیک و مورفولوژیک را در بر می‌گیرد. در سطح مولکولی و بیوشیمیایی، پاسخ‌های پیچیده‌ای از جمله تجمع اسمولیت‌های محافظ (مانند پرولین و گلیسین‌بتائین)، فعال‌سازی سیستم دفاع آنتی‌اکسیدانی برای خنثی‌سازی رادیکال‌های آزاد، بیان پروتئین‌های شوک حرارتی جهت حفاظت از پروتئین‌های سلولی و تنظیم شبکه‌های سیگنالینگ و بیان ژن‌ها رخ می‌دهد. این سازوکارها به‌طور هماهنگ، بقا و عملکرد گیاه را تحت تنش بهبود می‌بخشند. نتیجه‏گیری: مدیریت مؤثر تنش گرمایی مستلزم یک رویکرد یکپارچه است. راهکارهای زراعی مانند انتخاب ارقام متحمل، تاریخ کاشت مناسب، و مدیریت بهینه آبیاری پایه اصلی این رویکرد هستند. همچنین، کاربرد خارجی محافظ‌هایی مانند اسمولیت‌ها، فیتوهورمون‌ها (اسید آبسیزیک)، عناصر کم‌مصرف (مانند سلنیوم) و مواد مغذی می‌تواند آسیب ناشی از گرما را کاهش دهد. در نهایت، به‌کارگیری ابزارهای بیوتکنولوژیک و مولکولی برای درک عمیق‌تر سازوکارهای تحمل و توسعه ارقام مقاوم، راهکاری اساسی و آینده‌نگرانه است.

دانشگاه اراک سلول و بافت 2228-7035 16 4 2025 12 22 Investigation of antioxidant activity and seed oil fatty acid profile in diverse Okra cultivars بررسی ظرفیت آنتی ‫اکسیدانتی و ترکیب اسید چرب روغن بذر ارقام مختلف بامیه 421 434 733208 10.66224/JCT.16.4.421 FA سید مهدی طالبی دانشیار، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه اراک، اراک، ایران اسرا سلمان کارشناس ارشد، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه اراک، اراک، ایران پریسا مالکی کارشناس ارشد، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه اراک، اراک، ایران Journal Article Introduction: Okra (Abelmoschus esculentus (L.) Moench) is an annual herbaceous plant belonging to the family Malvaceae, valued for its high nutritional and medicinal properties. It is cultivated widely in various tropical and subtropical regions of the world, including many areas of Iran, where it serves as an important vegetable crop in local diets and traditional medicine. Aim: This study investigated the fatty acid composition and antioxidant activity of seed oils extracted from seven cultivars of Okra, namely White, Sultani, Red, Velvet, Green, Texas, and Fawn. Material and methods: Mature and intact seeds from each okra cultivar were powdered, and their oils were extracted using a modified Folch et al. method. The fatty acid methyl esters (FAMEs) were prepared from the extracted oils through transesterification. The resulting FAMEs were analyzed using gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS) apparatus to determine their fatty acid profiles. The antioxidant activity of the extracted oils was evaluated using the DPPH radical scavenging assay. Data were statistically analyzed using SPSS and MVSP software. Results: The seed oils of all okra cultivars contained both saturated and unsaturated fatty acids, with total proportions varying among the cultivars. The highest total saturated fatty acid content (42.53%) was observed in the White cultivar, while the lowest (36.41%) occurred in the Sultani cultivar. Palmitic acid was the predominant saturated fatty acid in all cultivars, with the highest (35.74%) and lowest (30.70%) levels detected in the White and Texas cultivars, respectively. Stearic acid was the second most abundant saturated fatty acid, ranging from 3.69% (Sultani) to 4.94% (White). Trace saturated fatty acids, including heptadecenoic, arachidic, behenic, and lignoceric acids were detected in all cultivars, whereas lauric acid was only found in a few cultivars (Velvet, Texas, and White cultivars). Unsaturated fatty acids constituted 57.33–63.33% of the total oil composition, with the lowest proportion in the White cultivar and the highest in the Sultani cultivar. Linoleic acid was the major unsaturated fatty acid in all cultivars except White, where oleic acid predominated. The highest linoleic acid content (11.40%) was recorded in the Red cultivar, and the lowest (8.25%) in White cultivar. In contrast, oleic acid content ranged from 18.20% (Red) to 49.30% (White). Trace amounts of other unsaturated fatty acids, including myristoleic, palmitoleic, trans-heptadecenoic, linolenic, gondoic, and erucic acids, were also detected across all cultivars. Additionally, eicosadienoic acid was present in trace amounts in all cultivars except White cultivar. Phytochemical analyses using the UPGMA dendrogram and Principal Component Analysis (PCA) grouped the cultivars into two main clusters, indicating significant phytochemical diversity. The predominance of oleic acid in the White cultivar, in contrast to linoleic acid in the others, along with the high palmitic acid content in the Velvet and White cultivars, contributed to the distinct placement of the White, Green, and Velvet cultivars from the remaining ones. The antioxidant activity, evaluated using the DPPH radical scavenging assay, revealed IC₅₀ values ranging from 689.23 µg/mL (Green cultivar) to 2551.12 µg/mL (Texas cultivar). Discussion: The variation in fatty acid composition among the seven Okra cultivars reflects significant biochemical diversity. Palmitic acid was the main saturated fatty acid in all cultivars, while oleic and linoleic acids dominated the unsaturated fractions. The White cultivar, with its high oleic acid level, exhibited greater oil stability potential, whereas other cultivars rich in linoleic acid offer higher nutritional value. Differences in fatty acid profiles may be related to cultivar-specific desaturase activity. The antioxidant activity varied notably, with the Green cultivar showing the strongest and the Texas cultivar the weakest radical scavenging capacity, likely due to variations in phenolic and unsaturated compounds. The UPGMA and PCA analyses confirmed chemical diversity and separated White, Green, and Velvet cultivars based on their distinct fatty acid compositions. هدف: بامیه گیاهی یک‫ساله خوراکی و با ارزش تغذیه ای و دارویی از خانواده مالواسه است که در مناطق مختلف جهان، از جمله ایران کشت می‫شود در این پژوهش، ترکیب اسیدهای چرب روغن و ظرفیت آنتی‫اکسیدانتی روغن حاصل از بذر هفت رقم مختلف بامیه خوراکی شامل ارقام سفید، سلطانی، قرمز، مخملی، سبز، تگزاسی و حنایی مورد بررسی قرار گرفت. مواد و روش‫ها: بذر های سالم و رسیده هر رقم پودر شده و سپس روغن آن‫ها به‫روش اصلاح شده Folch و همکاران استخراج شدند. سپس، اسید چرب موجود در روغن به متیل استر تبدیل شده و جهت شناسایی یه دستگاه‫های GS/MS و GC تزریق شدند. برای تعیین میزان ظرفیت آنتی‫اکسیدانتی از روش DPPH استفاده شد. داده‫های حاصل با نرم افزارهای MVSP و SPSS مورد تجزیه و تحلیل قرار گرفتند. نتایج: روغن این ارقام شامل طیفی از اسیدهای چرب اشباع و غیراشباع است که نسبت آن‌ها در میان ارقام مختلف تفاوت معنی‌داری دارد. بیشترین درصد مجموع اسیدهای چرب اشباع (53/42 درصد) در رقم بامیه سفید و کم‫ترین آن (41/36 درصد) در رقم سلطانی مشاهده شد. همچنین، بیش‫ترین (33/63 درصد) و کم‫ترین (33/57 درصد) میزان درصد مجموع اسیدهای چرب غیراشباع به‫ترتیب در ارقام سلطانی و سفید وجود داشت. اسید پالمتیک و اسید استئاریک به‌عنوان اسیدهای چرب اشباع غالب و اسیدهای لینولئیک و اولئیک به‌عنوان اسیدهای چرب غیراشباع اصلی شناسایی شدند. در رقم بامیه سفید، اسید اولئیک بیشترین سهم را در میان اسیدهای چرب غیراشباع داشت، در حالی‌‫که در سایر ارقام، اسید لینولئیک غالب بود. تجزیه خوشه‌ای به‫روش UPGMA و تحلیل مولفه‌های اصلی (PCA) منجر به تفکیک ارقام به دو گروه اصلی شد که نشان‌دهنده تنوع قابل توجه در ساختار شیمیایی روغن بذر بین ارقام است. همچنین، ظرفیت آنتی‌اکسیدانتی روغن‌های استخراج‌شده با استفاده از روش مهار رادیکال DPPH شناسایی و تعیین شد. میانگین IC50 روغن‌ها 67/1058 میکروگرم در میلی‌لیتر بود که از 23/689 میکروگرم در میلی‌لیتر در رقم سبز تا 12/2551 میکروگرم در میلی‌لیتر در رقم تگزاسی متغییر بود. بحث: ساختار روغن دانه بامیه رقم سفید بسیار متمایزتر از سایر ارقام است. یافته‌های این پژوهش می‌تواند در انتخاب ارقام مناسب جهت استخراج روغن برای اهداف تغذیه‌ای و صنعتی موثر واقع شود. همچنین،ساختار ترکیبات اسیدچرب غیر اشباع و ترکیبات فنولی بذر بر ظرفیت آنتی‫اکسیدانتی روغن بذر بامیه بسیار تاثیرگذار است.