scholarly journals Iron overload induced death of osteoblasts in vitro: involvement of the mitochondrial apoptotic pathway

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2611 ◽  
Author(s):  
Qing Tian ◽  
Shilei Wu ◽  
Zhipeng Dai ◽  
Jingjing Yang ◽  
Jin Zheng ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Iron Overload ◽  
Caspase 3 ◽  
Osteoblastic Cell ◽  
Ferric Ion ◽  
Oxygen Species ◽  
Mitochondrial Apoptosis ◽  
Apoptotic Pathway ◽  
Mitochondrial Apoptotic Pathway ◽  
Commercial Kits

BackgroundIron overload is recognized as a new pathogenfor osteoporosis. Various studies demonstrated that iron overload could induce apoptosis in osteoblasts and osteoporosis in vivo. However, the exact molecular mechanisms involved in the iron overload-mediated induction of apoptosis in osteoblasts has not been explored.PurposeIn this study, we attempted to determine whether the mitochondrial apoptotic pathway is involved in iron-induced osteoblastic cell death and to investigate the beneficial effect of N-acetyl-cysteine (NAC) in iron-induced cytotoxicity.MethodsThe MC3T3-E1 osteoblastic cell line was treated with various concentrations of ferric ion in the absence or presence of NAC, and intracellular iron, cell viability, reactive oxygen species, functionand morphology changes of mitochondria and mitochondrial apoptosis related key indicators were detected by commercial kits. In addition, to further explain potential mechanisms underlying iron overload-related osteoporosis, we also assessed cell viability, apoptosis, and osteogenic differentiation potential in bone marrow-derived mesenchymal stemcells(MSCs) by commercial kits.ResultsFerric ion demonstrated concentration-dependent cytotoxic effects on osteoblasts. After incubation with iron, an elevation of intracelluar labile iron levels and a concomitant over-generation of reactive oxygen species (ROS) were detected by flow cytometry in osteoblasts. Nox4 (NADPH oxidase 4), an important ROS producer, was also evaluated by western blot. Apoptosis, which was evaluated by Annexin V/propidium iodide staining, Hoechst 33258 staining, and the activation of caspase-3, was detected after exposure to iron. Iron contributed to the permeabilizatio of mitochondria, leading to the release of cytochrome C (cyto C), which, in turn, induced mitochondrial apoptosis in osteoblasts via activation of Caspase-3, up-regulation of Bax, and down-regulation of Bcl-2. NAC could reverse iron-mediated mitochondrial dysfunction and blocked the apoptotic events through inhibit the generation of ROS. In addition, iron could significantly promote apoptosis and suppress osteogenic differentiation and mineralization in bone marrow-derived MSCs.ConclusionsThese findings firstly demonstrate that the mitochondrial apoptotic pathway involved in iron-induced osteoblast apoptosis. NAC could relieved the oxidative stress and shielded osteoblasts from apoptosis casused by iron-overload. We also reveal that iron overload in bone marrow-derived MSCs results in increased apoptosis and the impairment of osteogenesis and mineralization.

scholarly journals Ginsenoside Rb1 Protects Neonatal Rat Cardiomyocytes from Hypoxia/Ischemia Induced Apoptosis and Inhibits Activation of the Mitochondrial Apoptotic Pathway

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Xu Yan ◽  
Jinwen Tian ◽  
Hongjin Wu ◽  
Yuna Liu ◽  
Jianxun Ren ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Neonatal Rat ◽  
Ginsenoside Rb1 ◽  
Caspase 9 ◽  
Apoptotic Pathway ◽  
Rat Cardiomyocytes ◽  
Mitochondrial Apoptotic Pathway ◽  
Neonatal Rat Cardiomyocytes ◽  
Hypoxia Ischemia

Aim. To investigate the effect of Ginsenoside Rb1 (GS-Rb1) on hypoxia/ischemia (H/I) injury in cardiomyocytesin vitroand the mitochondrial apoptotic pathway mediated mechanism.Methods. Neonatal rat cardiomyocytes (NRCMs) for the H/I groups were kept in DMEM without glucose and serum, and were placed into a hypoxic jar for 24 h. GS-Rb1 at concentrations from 2.5 to 40 µM was given during hypoxic period for 24 h. NRCMs injury was determined by MTT and lactate dehydrogenase (LDH) leakage assay. Cell apoptosis, ROS accumulation, and mitochondrial membrane potential (MMP) were assessed by flow cytometry. Cytosolic translocation of mitochondrial cytochrome c and Bcl-2 family proteins were determined by Western blot. Caspase-3 and caspase-9 activities were determined by the assay kit.Results. GS-Rb1 significantly reduced cell death and LDH leakage induced by H/I. It also reduced H/I induced NRCMs apoptosis induced by H/I, in accordance with a minimal reactive oxygen species (ROS) burst. Moreover, GS-Rb1 markedly decreased the translocation of cytochrome c from the mitochondria to the cytosol, increased the Bcl-2/ Bax ratio, and preserved mitochondrial transmembrane potential (ΔΨm). Its administration also inhibited activities of caspase-9 and caspase-3.Conclusion. Administration of GS-Rb1 during H/Iin vitrois involved in cardioprotection by inhibiting apoptosis, which may be due to inhibition of the mitochondrial apoptotic pathway.

scholarly journals Iron Overload Adversely Effects Bone Marrow Haematogenesis via SIRT-SOD2-MROS in a Process Ameliorated by Curcumin

2020 ◽  
Author(s):  
shujuan zhou ◽  
Lan Sun ◽  
Shanhu Qian ◽  
Yongyong Ma ◽  
Ruye Ma ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Iron Overload ◽  
Protective Effect ◽  
Suppressive Effect ◽  
Autophagic Cell Death ◽  
Iron Loading ◽  
Oxygen Species ◽  
Curcumin Treatment ◽  
Bone Marrow Damage

Abstract Background: Iron overload is common in patients with haematological disorders, and it is known to have a suppressive effect on haematogenesis. However, the mechanism by which iron overload affects haematogenesis is still unclear. The antioxidant curcumin has been reported to protect against iron overload-induced bone marrow damage, although the mechanism underlying this protective effect remains to be elucidated.Methods: We established iron overload cell and mouse models. Mitochondrion-derived reactive oxygen species (mROS) levels, autophagy levels, and the SIRT3/SOD2 pathway were examined in these models and in the bone marrow of patients with iron overload.Results: Iron overload was shown to depress haematogenesis and induce mitochondrion-derived superoxide anion-dependent autophagic cell death. Iron loading decreased SIRT3 protein expression, promoted an increase in SOD2, and led to the elevation of mROS. These effects were reversed by the overexpression of SIRT3. Curcumin treatment ameliorated peripheral blood cells, enhanced SIRT3 activity, decreased SOD2 acetylation, inhibited mROS production, and suppressed iron loading-induced autophagy.Conclusions: These results suggest that curcumin exerts a protective effect on bone marrow by reducing mROS-stimulated autophagic cell death in a manner dependent on the SIRT3/SOD2 pathway.

scholarly journals Mesenchymal Stem Cells from Rat Bone Marrow Downregulate Caspase-3-mediated Apoptotic Pathway After Spinal Cord Injury in Rats

2007 ◽  
Vol 32 (12) ◽  
pp. 2080-2093 ◽  
Author(s):  
Venkata Ramesh Dasari ◽  
Daniel G. Spomar ◽  
Craig Cady ◽  
Meena Gujrati ◽  
Jasti S. Rao ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Stem Cells ◽  
Bone Marrow ◽  
Spinal Cord Injury ◽  
Mesenchymal Stem Cells ◽  
Caspase 3 ◽  
Apoptotic Pathway ◽  
Cord Injury ◽  
Rat Bone

scholarly journals Iron overload adversely effects bone marrow haematogenesis via SIRT-SOD2-mROS in a process ameliorated by curcumin

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Shujuan Zhou ◽  
Lan Sun ◽  
Shanhu Qian ◽  
Yongyong Ma ◽  
Ruye Ma ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Iron Overload ◽  
Suppressive Effect ◽  
Autophagic Cell Death ◽  
Iron Loading ◽  
Oxygen Species ◽  
Mitochondrial Reactive Oxygen Species ◽  
Curcumin Treatment ◽  
Bone Marrow Damage

Abstract Background Iron overload, which is common in patients with haematological disorders, is known to have a suppressive effect on haematogenesis. However, the mechanism for this effect is still unclear. The antioxidant curcumin has been reported to protect against iron overload-induced bone marrow damage through an as-yet-unknown mechanism. Methods We established iron overload cell and mouse models. Mitochondrial reactive oxygen species (mROS) levels, autophagy levels and the SIRT3/SOD2 pathway were examined in the models and in the bone marrow of patients with iron overload. Results Iron overload was shown to depress haematogenesis and induce mitochondrion-derived superoxide anion-dependent autophagic cell death. Iron loading decreased SIRT3 protein expression, promoted an increase in SOD2, and led to the elevation of mROS. Overexpression of SIRT3 reversed these effects. Curcumin treatment ameliorated peripheral blood cells generation, enhanced SIRT3 activity, decreased SOD2 acetylation, inhibited mROS production, and suppressed iron loading-induced autophagy. Conclusions Our results suggest that curcumin exerts a protective effect on bone marrow by reducing mROS-stimulated autophagic cell death in a manner dependent on the SIRT3/SOD2 pathway.

Reactive oxygen species-independent rapid initiation of mitochondrial apoptotic pathway by chelerythrine

2011 ◽  
Vol 25 (8) ◽  
pp. 1581-1587 ◽  
Author(s):  
Takeshi Funakoshi ◽  
Toshihiko Aki ◽  
Haruka Nakayama ◽  
Yumi Watanuki ◽  
Satoko Imori ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Apoptotic Pathway ◽  
Mitochondrial Apoptotic Pathway

scholarly journals Moderate exercise attenuates caspase-3 activity, oxidative stress, and inhibits progression of diabetic renal disease in db/db mice

2009 ◽  
Vol 296 (4) ◽  
pp. F700-F708 ◽  
Author(s):  
S. Ghosh ◽  
M. Khazaei ◽  
F. Moien-Afshari ◽  
L. S. Ang ◽  
D. J. Granville ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Diabetic Nephropathy ◽  
Oxidative Damage ◽  
Renal Disease ◽  
Caspase 3 ◽  
Moderate Intensity ◽  
Caspase 8 ◽  
Apoptotic Pathway ◽  
Mitochondrial Apoptotic Pathway ◽  
Early Diabetic Nephropathy

Diabetic nephropathy, the leading cause of end-stage renal disease, is characterized by a proapoptotic and prooxidative environment. The mechanisms by which lifestyle interventions, such as exercise, benefit diabetic nephropathy are unknown. We hypothesized that exercise inhibits early diabetic nephropathy via attenuation of the mitochondrial apoptotic pathway and oxidative damage. Type 2 diabetic db/db and normoglycemic wild-type mice were exercised for an hour everyday at a moderate intensity for 7 wk, following which renal function, morphology, apoptotic signaling, and oxidative stress were evaluated. Exercise reduced body weight, albuminuria, and pathological glomerular expansion in db/db mice independent of hyperglycemic status. Changes in renal morphology were also related to reduced caspase-3 (main effector caspase in renal apoptosis), caspase-8 (main initiator caspase of the “extrinsic” pathway) activities, and TNF-α expression. A role for the mitochondrial apoptotic pathway was unlikely as both caspase-9 activity (initiator caspase of this pathway) and expression of regulatory proteins such as Bax and Bcl-2 were unchanged. Kidneys from db/db mice also produced higher levels of superoxides and had greater oxidative damage concurrent with downregulation of superoxide dismutase (SOD) 1 and 3. Interestingly, although exercise also increased superoxides, there was also upregulation of multiple SODs that likely inhibited lipid (hydroperoxides) and protein (carbonyls and nitrotyrosine) oxidation in db/db kidneys. In conclusion, exercise can inhibit progression of early diabetic nephropathy independent of hyperglycemia. Reductions in caspase-3 and caspase-8 activities, with parallel improvements in SOD expression and reduced oxidative damage, could underlie the beneficial effects of exercise in diabetic kidney disease.

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