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    Biological behaviors of muscarinic receptors in mesenchymal stem cells derived from human placenta and bone marrow
    (Mashhad Univ Med Sciences, 2020) Yegani, Arash Alizadeh; Maytalman, Erkan; Kozanoglu, Ilknur; Terzi, Menderes Yusuf; Aksu, Fazilet
    Objective(s): Cells perform their functional activities by communicating with each other through endogenous substances and receptors. Post-translation, stem cells function properly in new host tissue by carrying specific cell surface receptors. We aimed to characterize muscarinic receptor subtypes in mesenchymal stem cells (MSCs) together with osteogenic and adipogenic differentiation markers. Materials and Methods: mRNA levels of 5 muscarinic receptor subtypes (CHRM1 to 5), BMP-6, and PPAR gamma during osteogenic and adipogenic differentiation, under the effect of atropine blockade, were measured in MSCs obtained from human fetal membrane (FM) and bone marrow (BM). Additionally, the effect of atropine on differentiation in the 1st, 2nd, and 3rd passages of MSCs, obtained from human FM and BM, were analyzed by RT-qPCR. Results: CHRM1 mRNA levels increased in the FM group, while decreasing in the BM group. We found significant decreases in CHRM3 and CHRM5 mRNA levels in FM and BM groups, respectively. Atropine had variable effects based on cell source and receptor type. BMP-6 mRNA levels in differentiated osteogenic cells increased significantly compared to undifferentiated cells in both FM and BM groups. In MSCs derived from both sources, PPAR gamma mRNA levels in differentiated adipogenic cells increased significantly. Atropine showed no effect on MSCs differentiation. Conclusion: These results indicate that expressions of muscarinic receptors in MSCs derived from BM and FM can vary and these cells keep the potential of osteogenic and adipogenic differentiation in vitro. Besides, atropine had no effect on adipogenic and osteogenic differentiation of MSCs.
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    Effects of Curcumin on Iron Overload in Rats
    (Knowledge E, 2021) Ozbolat, Guluzar; Yegani, Arash Alizadeh
    Background: Iron overload, common in patients with hematological disorders, is a key target in drug development. This study investigated the effects of curcumin on iron overload in rats. Methods: Forty male Wistar rats weighing 139.78 +/- 11.95 gm (Mean +/- SD) were divided into three equal groups: (i) controls; (ii) iron overload group that received six doses of iron dextran 1000 mg/kg(--1) by intraperitoneal injections (i.p.); and (iii) iron overload curcumin group that received six doses of curcumin (1000 mg/kg BW by i.p.). In addition to six doses of iron dextran 1000 mg/kg(--1) by i.p., we studied the effects of curcumin on liver function enzymes (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]); antioxidant enzymes (malondialdehyde [MDA], total oxidant status [TOS], total antioxidant status [TAS]); hematological parameters (hemoglobin [Hb], hematocrit [Hct], red blood cells [RBC], white blood cells [WBC], mean corpus volume [MCV], mean corpuscular hemoglobin [MCH], mean corpuscular hemoglobin concentration [MCHC]); and iron parameters (serum iron profile, transferrin, total iron-binding capacity [TIBC], ferritin, and transferrin saturation [TS%]). Results: Curcumin caused a significant decrease in the Hct and Hb concentrations in Group III (P < 0.05). It also significantly reduced the serum levels of ALT (52.45 +/- 4.51 vs 89.58 +/- 4.65 U/L) and AST (148.03 +/- 6.47 vs 265.27 +/- 13.02 U/L) at the end of the study (P < 0.05). The TIBC, transferrin levels, and TS significantly decreased when the rats were administered curcumin serum iron (P < 0.05). The TAS level significantly increased in Group III in comparison to Group I (the control group) (P < 0.05). At the end of the study, curcumin significantly reduced the serum levels of TOS (12.03 +/- 2.8 vs 16.95 +/- 5.05 mmol H2O2/L) while the TAS (1.98 +/- 0.42 vs 1.06 +/- 0.33 mmol Trolox equiv./L) was increased. Conclusion: The findings of the present study suggest the therapeutic potential of curcumin against iron overload.
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    In vitro effects of iron chelation of curcumin Fe (III) complex
    (Çukurova Üniversitesi, 2019) Özbolat, Gülüzar; Yegani, Arash Alizadeh
    Purpose: The aim of this study was to investigate the cytotoxicity effect, iron chelator and antioxidant activities of iron (III) ions with curcumin ligand that may be used in the treatment of iron overload. Materials and Methods: The cytotoxic activities of the ligand and the complex were evaluated by the MTT assay. The SOD activity of the complex of curcumin was determined by using its ability to inhibit the reduction of NBT. The catalytic activity studies of Fe(III) complex in DMSO towards the disproportionation of hydrogen peroxide were also performed. Results: The IC50 values are found in 6.8 μM catalase activity was measured. Where at a concentration of 2.0 mM, the activity was equivalent to 183.30 U/L. The complex shows a catalase activity. The complex showed minimal toxicity. IC50 values found 5.3 mg/ml. The observed cytotoxicity could be pursued to obtain a potential drug. The iron chelator effects were determined by Ferrozine reagent. Curcumin, the most active extract interfered with the formation of ferrous and ferrozine complex. It demonstrated strong chelating activities. The result showed that the complexes possess considerable SOD activity. This finding indicates that the iron complex is capable of removing free radicals. Conclusion: The study results revealed that the iron(III) complex of curcumin with an appropriate potential drug may act as a protector against oxidative stress. Therefore, all results suggest that curcumin may represent a new approach in the treatment of iron overload.
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    Synthesis, characterization and electrochemistry studies of iron(III) complex with curcumin ligand
    (Wiley, 2018) Ozbolat, Guluzar; Yegani, Arash Alizadeh; Tuli, Abdullah
    Iron overload is a serious clinical condition for humans and is a key target in drug development. The aim of this study was to investigate the coordination of iron(III) ions with curcumin ligand that may be used in the treatment of iron overload. Iron(III) complex of curcumin was synthesized and structurally characterized in its solid and solution state by FT-IR, UV-Vis, elemental analysis, and magnetic susceptibility. Electrochemical behaviour of the ligand and the complexes were examined using cyclic voltammetry. The cytotoxic activities of the ligand and the iron(III) complex were evaluated by the MTT assay. Curcumin reacted with iron in high concentrations at physiological pH at room temperature. Subsequently, a brown-red complex was formed. Data regarding magnetic susceptibility showed that the complexes with a 1:2 (metal/ligand) mole ratio had octahedral geometry. The complex showed higher anti-oxidant effect towards the cell line ECV304 at IC50 values of 4.83 compared to curcumin. The complex exhibited very high cytotoxic activity and showed a cytotoxic effect that was much better than that of the ligand. The potentials for redox were calculated as 0.180V and 0.350V, respectively. The electrochemistry studies showed that Fe3+/Fe2+ couple redox process occurred at low potentials. This value was within the range of compounds that are expected to show superoxide dismutase activity. This finding indicates that the iron complex is capable of removing free radicals. The observed cytotoxicity could be pursued to obtain a potential drug. Further studies investigating the use of curcumin for this purpose are needed.
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    Synthesis, characterization, biological activity and electrochemistry studies of iron(III) complex with curcumin-oxime ligand
    (Wiley, 2020) Ozbolat, Guluzar; Yegani, Arash Alizadeh
    Iron overload is a key target in drug development. This study aimed to investigate the coordination of Fe(III) ions with a curcumin-oxime ligand that may be used in the treatment of iron overload. The synthesis of the curcumin-oxime ligand and curcumin-oxime-Fe(III) complex was successfully made and characterized in its solid-state and solution-state using FT-IR, UV-Vis, elemental analysis, and H-1-NMR. However, in this study, we investigated the apoptotic effects of the curcumin-oxime Fe (III) complex on SW480. SW480 cells were exposed to 99.2% medium for 48 hours. After 48 hours, the incubation period, cells were harvested by centrifugation and washed in phosphate-buffered saline (PBS) and lysed in radio-immunoprecipitation assay (RIPA) buffer for 20 minutes and supernatants were taken and pellets were discarded. ELISA test was used to examine the expression, and activity of cleaved caspase-3, Bax, and Bcl-2 proteins in SW480 cells. ELISA test results indicated that the activities of apoptotic proteins Bax, caspase 3 and Bcl-2 in human SW480 cell lines significantly increased in 48 hours treatment. Also, the activity of Bcl-2 was observed to decrease significantly. Catalase activities of the complex were investigated. The findings showed that the complex has a catalase activity. The findings suggest that this type of complex may constitute a new and interesting basis for the future search of new and more potent drugs. The SOD activity of the result showed that the complexes possessed a considerable SOD activity with an IC50 value of 7.685 mu M. Also, when compared with the control, a complex increased the SOD levels (P < .05). Electrochemistry studies in the literature have shown that the Fe3+/Fe(2+)couple redox process occurs in low potential. This value is within the range of compounds that are expected to show superoxide dismutase activity. The I-pc/I(pa)shows that one electron transport takes place in the complex. Our results suggest that curcumin-oxime may represent a new approach in the treatment of iron overload.