Browsing by Autor "Hossein Maghsoudi"

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Biochemical Chgaracterization of Recombinant Baculovirus 373 K/E p53 Protein
(2018) Hossein Maghsoudi; Uttam Pati
In this study, we expressed and purified the recombinant baculovirus 373 K/E p53 protein in a baculovirus expression system to characterize this mutant and compare it with wild type p53. Gel- filtration chromatography and chemical cross-linking experiments indicated that purified recombinant baculovirus 373 K/E p53 protein assembles into multimeric forms ranging from tetramers to polymers. Gel-mobility shift assays and protein-DNA cross-linking studies demonstrated that the recombinant protein binds, to a consensus DNA target as a dimer but that additional p53 mutant molecules may then associate with the preformed p53-dimer-DNA complexes to form a larger p53_DNA complexes. These observations suggest that the p53 mutant tetramers and polymers that forms the minimal p53 mutant complex in solution dissociated upon DNA binding to form p53 mutant dimmer DNA complexes. The DNA binding activity of this mutant was then investigated using electrophoretic mobility shift assays as well as supershift assay with anti-p53 antibodies. Binding of the anti-p53 antibody PAb421to the oligomerization promoting domain on p53 stimulated the sequential formation of both the p53_dimer DNA and larger p53-DNA complexes
Evaluating the Effects of Carnosic acid on TNF-α Gene Expression in LPS-activated Synoviocytes
(Medical University of Ilam, 2018) Mehranoush Saffarpour; Hossein Maghsoudi; Mohammad Fazilati; Mohammad Fazilati
Introduction: Osteoarthritis (OA) is a major cause of disability among adults. As a progressive degenerative joint disorder, OA is characterized by cartilage damage, changes in the subchondral bone and inflammation of the synovium tissue and tendon. Non-pharmacologic agents modulating pro-inflammatory mediator expression offer considerable promise as safe and effective treatments for OA. The aim of this study was to evaluate the effects of Carnosic acid (CA) on TNF-α gene expression in synoviocytes. Material & methods: To determine the dose response of CA, synoviocytes (5 × 105 cells/well) were incubated at 5% CO2, 37ºC for 72 h with (1) control media alone or (2) CA at concentrations of 0.01, 0.09, 0.1, 0.9, 1, 9,10and 90 mg/ml. Synoviocytes were treated with LPS (20 ng/ml) for 1h and TNF-α mRNA were measured by both reverse-transcriptase PCR and Real-time PCR. Findings: CA reduced TNF-a, expression in LPS-activated synoviocytes to levels similar to nonactivated control levels. Cells activated with 20 ng/ml LPS showed a significant upregulation of TNF-α expression. In activated synoviocytes cells pretreated with CA, TNF-α was reduced by 48/11% when compared to activated control cells. Discussion & conclusions: This study demonstrates that CA acts as a potent inhibitor of TNF-α. Previous research has shown that these mediators are important in the pathogenesis of OA and reduction in these mediators has been associated with amelioration of cartilage breakdown.
The Effect of Hydroalcoholic Extract of Echinops cephalotes L. on Expression of Inflammatory Factors in Chondrocytes and THP-1 Cells as a Model of Monocyte/Macrophage and Human Cartilage Cells in Osteoarthritis
(2018) Mahdi Taherian; Hossein Maghsoudi; Reza Taherian; Hossein Rastegar
Background: Osteoarthritis (OA) is one of the main causes of physical disability worldwide. Considering the complications of common treatments of OA, including non-steroidal anti-inflammatory drug (NSAIDs) and corticosteroids, establishment of new treatments is crucial. Objective: This study aimed to explore the effect of Echinops cephalotes extract on the main inflammatory biomarkers in OA. Methods: Hydroalcoholic extract of Echinops cephalotes, Ibuprofen and betamethasone were prepared to investigate their effects on inflammatory biomarkers. Human monocyte/macrophage (THP-1) cells and chondrocytes cells were used as a model of monocyte/macrophage and human cartilage cells in osteoarthritis. Lipopolysaccharide (LPS) was used to induce production of inflammatory cytokines in both cells. After RNA extraction and production of cDNA, RT_PCR & PCR were done. Then Real Time-PCR was used to investigate the amount of expression of proinflammatory genes. Results: Echinops cephalotes extract reduced mRNA expression level of proinflammatory cytokines in the cells induced by LPS. Moreover, production of PGE2 and NO in in the LPS-induced THP-1 cells was reduced by this extract. Ibuprofen and betamethasone were more effective in reducing above inflammatory agents than the extract. Conclusion: Echinops cephalotes extract can be used as a supplementary treatment option in osteoarthritis to reduce NSAIDs and corticosteroids dose in treatment of this disease.