Objective(s): Fluvoxamine is a well-known selective serotonin reuptake inhibitor (SSRI); Despite its anti-inflammatory effect, little is known about the precise mechanisms involved. In our previous work, we found that IP administration of fluvoxamine produced a noticeable anti-inflammatory effect in carrageenan-induced paw edema in rats. In this study, we aimed to evaluate the effect of fluvoxamine on the expression of some inflammatory genes like intercellular adhesion molecule (ICAM1), vascular cell adhesion molecule (VCAM1), cyclooxygenases2 (COX2), and inducible nitric oxide synthase (iNOS). Materials and Methods: An in vitro model of LPS stimulated human endothelial cells and U937 macrophages were used. Cells were pretreated with various concentrations of fluvoxamine, from 10-8 M to 10-6 M. For in vivo model, fluvoxamine was administered IP at doses of 25 and 50 mg/kg-1, before injection of carrageenan. At the end of experiment, the expression of mentioned genes were measured by quantitative real time (RT)-PCR in cells and in paw edema in rat. Results: The expression of ICAM1, VCAM1, COX2, and iNOS was significantly decreased by fluvoxamine in endothelial cells, macrophages, and in rat carrageenan-induced paw edema. Our finding also confirmed that IP injection of fluvoxamine inhibits carrageenan-induced inflammation in rat paw edema. Conclusion: The results of present study provide further evidence for the anti-inflammatory effect of fluvoxamine. This effect appears to be mediated by down regulation of inflammatory genes. Further studies are needed to evaluate the complex cellular and molecular mechanisms of immunomodulatory effect of fluvoxamine.

The interaction between immune cells and endothelial lining of blood vessels is vital in many processes such as inflammatory and immune responses as well as cancer cell metastasis. The expression level of VCAM-1 is regulated by many factors including the promoter activity that is possibly affected by the single nucleotide polymorphisms (SNPs) present in the promoter. There are previous reports suggesting an important role for rs3783605 at -420 position in the pathogenesis of VCAM1-associated diseases. This is possibly due to the effect of this SNP on promoter activity and gene expression. Therefore, present study was designed to investigate the effect of rs3783605 on the activity of VCAM-1 gene promoter in human umbilical vein endothelial cells (HUVEC).In this study, two appropriate expression vectors containing VCAM1 promoter with different alleles of rs3783605 were constructed to express the Green Fluorescent Protein (GFP). Expression vectors were transfected into HUVECs and their EGFP expression level was assessed by the fluorescent microscopy and real-time PCR.Bright green fluorescence was seen in the HUVECs transfected by expression vector containing CMV promoter. The expression level in the cells transfected by vector containing promoter with A allele of rs3783605 was 0.14888 folds and G allele was about 0.37851 folds of cells transfected by vector having CMV promoter (p<0.001). Moreover, HUVECs transfected by G allele of rs3783605 showed about 2-fold higher transcriptional activity compared with the A allele, (p=0.049).Our findings showed that rs3783605 polymorphism may play a role in VCAM-1 gene expression. Therefore, it is likely that it may have an important role in the pathogenesis of VCAM1-associated diseases and tumor metastases.

Infertility is known as one of the most common problems among couples. In this regard, generation of male germ cells from adult stem ones are among the current promising priorities of researchers. Mesenchymal stromal cells (MSCs) were previously induced to differentiate into germ-like progenitors in vitro. Monophosphoryl lipid A (MPLA) is a detoxified derivative of lipopolysaccharides (LPS) that lacks many of the endotoxic properties of LPS. Our present study aimed to investigate the expression of migration genes (CXCR4, VCAM1, VEGF, MMP2, and VLA4), and differentiation markers during human umbilical mesenchymal stromal cells (hUMSCs) culture in the presence of retinoic acid (RA) and MPLA-treated acellular testis. Accordingly, the high expression levels of deleted in azoospermia-like (DAZL), piwi-like RNA-mediated gene silencing 2 (PIWIL2) transcripts as well as protein were consequently observed in treated hUMSCs. It was concluded that combination treatment (i. e., MPLA/RA) had more prominent results than each of the treatments alone, even though MPLA and RA could be regarded as inducer of migration and differentiation, respectively. Ultimately, it was suggested to introduce the use of combination treatment as a more effective strategy to improve therapies in regenerative medicine.

Background and objectives: Rutin is a lipophilic natural flavonoid. It is found in vegetables, citrus fruits, and beverages. This study aims to evaluate rutin metabolic pathways in human senescent stromal cells. Methods: Data are extracted from the Gene Expression Omnibus (GEO) database and pre-evaluated via GEO2R to find the significant differentially expressed genes (DEGs). The significant DEGs were assessed via protein-protein interaction PPI network analysis to explore the hub genes. The hubs were screened via directed PPI to find the critical DEGs. Results: Data were assessed via volcano plot, Uniform Manifold Approximation and Projection (UMAP) plot, and Venn diagram. A total number of 9124 significant DEGs were analyzed to determine 33 upregulated and 61 downregulated hubs. The identified hubs were investigated via directed PPI and Il1B. ICAM1, CCL2, EGF, CXCL8, PTGS2, CAMK2B, CCN2, VCAM1, ELN, CXCL12, BGN, and TLR4 were pointed out as critical hub genes. Conclusion: Il1B, CCL2, GNAO1, ICAM1, EGF, and CXCL8 appeared as controller genes affected by rutin while PTGS2 and CAMK2B were the most controlled individuals. The finding refers to the significant advantages of the rutin effect on the function of treated cells. These advantages are corresponded to the usefulness of rutin as an herbal drug candidate. However, more investigations are required to decrease its side effects.

Objectives: The aim of the present study was to isolate MSCs using either low or high cell-seeding density culture system and to compare the cells produced in two systems in terms of their morphology, differentiation potential as well as the expression of some cell surface antigens.Materials and Methods: The bone marrow cells from Balb/c mice of 6-8-weeks old were cultivated at 2.5×106 cells/cm2. Primary culture was then tripsinized, cultured either at 50 cells/cm2 as low density culture systems or at 8×104 cell/cm2 as high density culture system. The passaged-3 cells from either system were further examined and compared in terms of morphology, differentiation potential and the expression of some surface antigens including CD135, CD44, CD31, Thy1.2, CD11b, CD45, CD34, Vcam1, Sca-1, and c-Kit.Results: In contrast to high–density culture system, in low density one, a few colons were produced and the culture contained more fibroblastic cells. According to the differentiation results, in contrast to high density system, more percentage of the cells being produced in low density culture system has been differentiated into bone and adipose cells. Furthermore, CD135, CD34, CD31 and Vcam were not appeared on the cells of low density system and Thy 1.2 surface antigen was not expressed on the cells of high density culture system. All these differences were statistically significant.Conclusions: Taken together, it seems that low density culture system is an appropriate method to isolate and expand murine mesenchymal stem cells.

Introduction: Pluripotent stem cells have been used by various researchers to differentiate and characterize endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) for the clinical treatment of vascular injuries. Studies continue to differentiate and characterize the cells with higher vascularization potential and low risk of malignant transformation to the recipient. Unlike previous studies, this research aimed to differentiate induced pluripotent stem (iPS) cells into endothelial progenitor cells (EPCs) and VSMCs using a step-wise technique. This was achieved by elucidating the spatio-temporal expressions of the stage-specific genes and proteins during the differentiation process. The presence of highly expressed oncogenes in iPS cells was also investigated during the differentiation period. Methods: Induced PS cells were differentiated into lateral mesoderm cells (Flk1 + ). The Flk1 + populations were isolated on day 5. 5 of the mesodermal differentiation period. Flk1 + cells were further differentiated into EPCs and VSMCs using VEGF165 and platelet-derived growth factor-BB (PDGFBB), respectively, and then characterized using gene expression levels, immunocytochemistry (ICC), and western blot (WB) methods. During the differentiation steps, the expression levels of the marker genes and proto-oncogenic Myc and Klf4 genes were simultaneously studied. Results: The optimal time for the isolation of Flk1 + cells was on day 5. 5. EPCs and VSMCs were differentiated from Flk1 + cells and characterized with EPC-specific markers, including Kdr, Pecam1, CD133, Cdh5, Efnb2, Vcam1,and VSMC-specific markers, including Acta2, Cnn1, Des, and Myh11. Differentiated cells were validated based on their temporal gene expressions, protein synthesis, and localization at certain time points. Significant decreases in Myc and Klf4 gene expression levels were observed during the EPCs and VSMC differentiation period. Conclusion: EPCs and VSMCs were successfully differentiated from iPS cells and characterized by gene expression levels, ICC, and WB. We observed significant decreases in oncogene expression levels in the differentiated EPCs and VSMCs. In terms of safety, the described methodology provided a better safety margin. EPCs and VSMC obtained using this method may be good candidates for transplantation and vascular regeneration.