Seyedeh Tahereh Haeri, Mohammad Ali Azarbayjani, Maghsoud Peeri,
Volume 13, Issue 4 (7-2019)
Abstract
Background and Objectives: Apoptosis is essential for the survival and normal functioning of multicellular organisms, yet any interruption in this process could be detrimental. Increased production of reactive oxygen species and oxidative stress are key factors affecting apoptosis. Our objective was to determine the impact of exercise with and without vitamin D supplementation on expression of FLIP, Fas, and caspase 8 in lung of rats poisoned with H2O2.
Methods: Forty-eight adult male rats were randomly divided into six groups: (C), (H), (HD), (HE), (HDE) and dimethyl sulfoxide. Groups H, HE, HD and HDE received 1 mmol/Kg intraperitoneal injection of H2O2. HE and HDE groups ran on treadmill for eight weeks. Expression of FLIP, Fas and caspase 8 was measured in lung tissues using RT-qPCR. Statistical analysis of data was carried out using SPSS 22 at significance level of 0.05.
Results: Vitamin D supplementation caused a significant decrease in expression of Fas (P=0.014) and caspase 8 (P=0.016) compared to the control group. However, it significantly overexpressed FLIP in the lung tissues compared to the control group (P=0.005). Exercise with and without vitamin D supplementation had no significant effect on the expression of the apoptosis regulatory genes.
Conclusion: Our results show that VD exerts protective effects on lung tissue by regulating apoptotic factors. Aerobic exercise alone and combined with VD has no significant effect on the apoptotic factors. These results indicate that VD supplementation can reduce lung injury under oxidative stress conditions.
Ali Salehi, Hajar Abaszadeh, Parvin Farzanegi,
Volume 13, Issue 4 (7-2019)
Abstract
Background and objectives: Pyruvate dehydrogenase kinase 4 (PDK4) is an enzyme involved in the regulation of glucose and fatty acid metabolism and homeostasis. The purpose of this study was to investigate effects of continuous exercise and resveratrol supplementation on insulin resistance and expression of PDK4 in the soleus muscle of elderly rats with type 2 diabetes.
Methods: After inducing diabetes in 42 old male rats (mean weigh: 250-300 g, aged about two years), the rats were randomly divided into six groups: healthy control, diabetic control, continuous exercise, supplementation, continuous exercise + supplementation and sham. The continuous exercise group was subjected to eight weeks of running, five sessions a week. Daily 20 mg/Kg body weight resveratrol supplementation was administered to the supplementation and supplementation + exercise groups. To evaluate insulin resistance, the HOMA-IR insulin resistance index was calculated as the product of insulin (μIU/mL) and glucose (mmol/L) divided by 22.5. Expression of the PDK4 gene in the soleus muscle was analyzed by RT-PCR. Data were analyzed with IBM SPSS Statistics 23 using one-way ANOVA and Tukey's post hoc test at a significance level of 0.05.
Results: Resveratrol alone or combined with continuous exercise significantly increased PDK4 expression and slightly decreased insulin resistance in the subjects.
Conclusion: Our results show that diabetes is associated with reduced PDK4 expression and increased insulin resistance. These effects can be effectively reversed in the soleus muscle of elderly diabetic rats by resveratrol supplementation alone or combined with continuous exercise.
Reza Bagheri, Samane Darroudi, S Mojtaba Hosseini, Hossein Nikkar, Fatemeh Khodadadi, Somayye Kasraee, Mahbube Sazegar, Amir Rashidlamir,
Volume 14, Issue 3 (5-2020)
Abstract
Background and objectives: Atherosclerotic cardiovascular disease is the leading cause of death in industrialized countries. High level of plasma lipids including cholesterol and triglycerides is one of the most important risk factors of atherosclerosis. Previous studies have shown that three members of the ATP-binding cassette, subfamily G (ABCG4, ABCG5 and ABCG8) are involved in transporting sterols across membranes. The purpose of this study was to assess the effect of high-intensity resistance training (HIRT) and aerobic exercise (AE) on expression of the ABCG4, ABCG5 and ABCG8 genes in female athletes.
Methods: Twenty-four female athletes from the Khorasan Province (Iran) were randomly selected and assigned into three groups: control (N=8), AE (N=8) and HIRT (N=8). The subjects in the HIRT group and the AE group performed exercise at intensity of 75-80% of 1-repetion maximum and 75-80% of maximum heart rate, respectively. Blood samples were collected at baseline and immediately after the exercise session. After isolation of lymphocytes by centrifugation and purification of mRNA, gene expression changes were investigated by Real-Time-PCR. Data were analyzed using one-way ANOVA and the Tukey's test.
Results: Both training protocols significantly increased the expression of ABCG4, ABCG5 and ABCG8 in the subjects (P<0.05), and there was no significant difference in the expression of these genes between the experimental groups (P>0.05).
Conclusion: A single session of HIRT and AE may have beneficial effects on prevention of atherosclerosis.
Mohammad Arefi, Abbas Abdollahi, Ayyoob Khosravi, Abdolavahab Moradi, Seyed Hamid Aghaee-Bakhtiari, Naimeh Javid, Mehdi Evazalipour, Anvarsadat Kianmehr,
Volume 15, Issue 2 (3-2021)
Abstract
Background and objectives: Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality in the world. MicroRNAs (miRNAs) have potential as diagnostic biomarkers for various diseases including cancer. This study was undertaken to investigate expression of miR-21 before and after surgery in patients with hereditary CRC.
Methods: After collecting blood samples from 39 patients and 39 healthy controls, total RNA was extracted by the TRIzol method. Following cDNA synthesis, expression of miR-21 in serum of subjects was evaluated using real-time PCR, along with two reference genes, let-7d and let-7g. The real-time expression results and Ct values were collected and analyzed based on the 2-∆∆ct method.
Results: In spite of tumor removal, serum miR-21 expression levels was significantly higher in hereditary CRC patients compared with controls (P=0.022).
Conclusion: Our results confirmed that samples from hereditary cases of CRC must not be included in experiments on the diagnostic potential of miRNAs.
Arash Nikyar, Azam Bolhassani, Fatemeh Rouhollah, Masoumeh Heshmati,
Volume 15, Issue 2 (3-2021)
Abstract
Background and objectives: HIV-1 Nef and Vpr antigens have been described as suitable candidates for therapeutic HIV vaccine development. The aim of this study was to generate Nef-Vpr fusion gene construct and to clone the construct into pET-23a (+), a prokaryotic expression vector.
Methods: HIV-1 Nef and Vpr genes were PCR-amplified from the pNL4-3 plasmid using specific primers and Pfu DNA polymerase. Results of PCR amplification were visualized by electrophoresis on 0.8% agarose gel. At first, the amplified Nef fragment was cloned into NheI and BamHI restriction sites of pET-23a expression vector. Next, cloning of Vpr gene was performed into BamHI and HindIII restriction sites of the pET-23a-Nef vector. Finally, purity of the recombinant pET-23-Nef-Vpr construct was determined by NanoDrop spectrophotometry.
Results: PCR amplification of Nef and Vpr genes was confirmed by detection of ~ 620 bp and ~ 291 bp bands, respectively. Cloning of the Nef-Vpr construct into the vector was confirmed by detection of a ~ 911 bp fragment following enzymatic digestion with NheI and HindIII and sequencing.
Conclusion: The successful construction of recombinant fusion plasmid encoding a chimeric Nef-Vpr gene was performed in a prokaryotic expression vector for development of HIV-1 recombinant protein vaccine in near future.
Shahram Zehtabian, Reza Alibakhshi, Seyed Yousef Seyedena, Ali-Reza Rai,
Volume 15, Issue 5 (9-2021)
Abstract
Background and objectives: Coronary artery disease (CAD) refers to stenosis or obstruction of coronary artery due to atherosclerosis or clotting. The aim of this study was to evaluate possible association of serum miRNA-197 (miR-197) expression as a biomarker for CAD diagnosis.
Methods: In this study, 100 patients with CAD who had angiography and vascular transplantation were selected. Expression of miR-197 was evaluated using real-time RT-PCR technique and the SYBR Green method. The Pearson's correlation coefficient was used to determine relationship of miR-197 expression and severity of coronary artery disease. The t-test was used to determine significance of expression of miR-197 in the study groups. All statistical analyses were carried out in SPSS 16 and at significance of 0.05.
Results: The results showed a direct relationship between miR-197 expression and CAD severity. The relative expression of miR-197 in the CAD patients was significantly higher than that in control subjects (P<0.004).
Conclusion: It seems that miR-197 can be considered as an indicator of coronary endothelial cell function. This microRNA could be used as a biomarker for CAD prognosis and treatment progression.
Nakisa Zarrabi Ahrabi, Hedyeh Ghadiripour, Seyed Mehdi Tabaie,
Volume 16, Issue 5 (9-2022)
Abstract
Background and objectives: Human Dermal Fibroblasts (HDF) are involved in the production of the extracellular matrix, formation of the connective tissue, and wound healing. Considering the role of ciprofloxacin in the treatment of skin infections and the subsequent oxidative stress as well as the protective effects of vitamin E and low power laser against inflammation and oxidative stress, we evaluated combined effects of low power laser and vitamin E on inflammation and oxidative stress in HDF cells treated with ciprofloxacin.
Methods: Morphology of the cells was studied using an inverted microscope. Viability of the cells was assessed using the MTT assay, and the concentration of reactive oxygen species was determined after exposure of the cells to ciprofloxacin (5, 25, 50, 75, and 100 μg/ml), vitamin E (1 mg/ml), and low power laser (660 nm; power density: 30 mW.cm−2).
Results: The survival rate of the cells increased significantly after the treatment with ciprofloxacin, vitamin E, and low power laser compared with the cells treated with ciprofloxacin and vitamin E (p<0.001). The amount of reactive oxygen species increased in the treated cells when compared with those only treated with ciprofloxacin and vitamin E.
Conclusion: The low power laser treatment has favorable effects on the growth of HDF cells, which can be beneficial for wound healing, even in the presence of ciprofloxacin.
Darshana Kottahachchi, Tharushika Deshani Hewapathirana, Thisali Chandula Perera, Shashikala Suresh,
Volume 18, Issue 2 (3-2024)
Abstract
Multiple myeloma (MM) is a plasma cell neoplasm that is characterized by the clonal proliferation of malignant plasma cells in the bone marrow. It is considered the second most common hematological malignancy which accounts for approximately 1% - 2% of all cancers and among 10% of hematological malignancies. Autologous peripheral blood stem cell Transplantation (PBSCT) is the best treatment for MM. Since the optimum harvested stem cell yield is a crucial factor for sufficient engraftment, the enumeration of Mononuclear cell (MNC) count in peripheral blood and harvested CD 34+ stem cell count can be considered as the best predictive markers for the best timing of apheresis which positively correlates with engraftment outcome of PBSCT.
MNC count can be obtained using either a hematological analyzer or peripheral blood smear while flow cytometry is the advanced technology that can be used to enumerate CD 34+ stem cell count other than peripheral blood smear. The unavailability of a flow cytometer, the expensiveness of this method, and the lack of trained personnel regarding this new technology, especially in lower-middle-income countries cause disturbance in the enumeration of stem cells. In such a situation, this review describes the importance of establishing an association between peripheral blood MNCs and harvested CD 34+ cells. Furthermore, this association facilitates conducting effective PBSCT for MM patients even in the absence of a flow cytometer and eventually, it focuses on decentralizing the treatment of PBSCT.
Afrooz Daneshparvar , Iman Jamhiri , Vahid Razban, Jafar Fallahi , Nasrin Hamidizadeh , Behnam Moghtaderi , Mehdi Dianatpour ,
Volume 18, Issue 5 (9-2024)
Abstract
Background: A rare heterozygous DYRK1B mutation (R102C) recently linked to a familial form of metabolic syndrome prompted this study to introduce the R102C mutation into the mouse DYRK1B gene, utilizing recombinant lentiviruses for long-term gene expression.
Methods: In the present fundamental study, the DYRK1B R102C mutation was generated via Overlap Extension-PCR (OE-PCR) and inserted into the LeGO-iG2 transfer vector with a GFP marker. Recombinant lentiviruses were produced by co-transfection of the transfer vector carrying DYRK1B R102C, psPAX2 (Packaging vector), and pMD2 (Envelope vector) into HEK-293T cells.
Results: The accuracy of the intended mutation was confirmed through OE-PCR and sequencing. Expression of DYRK1B and successful gene transfer were visualized using a fluorescence microscope to detect the GFP marker. Lentiviral titer was quantified using flow cytometry, with an infection efficiency of 108 TU/ml in HEK-293T cells.
Conclusion: DYRK1B plays a crucial role in the pathogenesis of metabolic syndrome, central obesity, early-onset coronary artery disease, hypertension, type 2 diabetes, and adipogenesis, suggesting its potential as a target for therapeutic interventions. Lentiviruses carrying the DYRK1B R102C mutation offer significant advantages for both in vitro and in vivo research on metabolic syndrome. This study showcases the successful application of recombinant lentiviral vectors for gene transfer into eukaryotic cells.