Related Experiment Video
Updated: Jul 19, 2026
Caspase-3 Activity in the Rat Amygdala Measured by Spectrofluorometry After Myocardial Infarction
Published on: January 12, 2016
Effect of Ghrelin on Caspase 3 and Bcl2 Gene Expression in H2O2 Treated Rat's Bone Marrow Stromal Cells
Alireza Abdanipour1, Masoud Dadkhah2, Mohsen Alipour2
1Department of Anatomical Sciences, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Abstract:
The antiapoptotic effect of ghrelin in various cell lines including bone marrow stromal cells (BMSCs) has been proved. However, the real mechanism of this effect is not clear. Caspase3 and Bcl2 are well-known pro- and antiapoptotic regulatory genes in eukaryotes. The aim of the study was to find out the effect of ghrelin on Caspase 3 and Bcl2 change in BMSCs. Rat BMSCs were cultivated in DMEM. Passage 3 BMSCs were treated with ghrelin 100 μM for 48 h. Real-time PCR for Caspase 3 and Bcl2 was carried out from B (untreated BMSCs), BH (BMSCs treated with 125 µM H2O2), BGH (BMSCs treated with 100 µM ghrelin then 125 µM H2O2) and BG (BMSCs treated with 100 µM ghrelin) groups. For immunofluorescence, cells were incubated with anti Caspase 3 and Bcl2monoclonal antibodies. Primary antibodies were visualized using the FITC method. All data are presented as means ± SEM. Values of P<0.05 were considered statistically significant. Ghrelin decreased mRNA expressions of Caspase-3 significantly as compared to the BH group (P<0.05). Also, Bcl-2 gene expression showed an increment in BG group as compare with BH and BGH groups (P<0.05). A high present of Bcl-2 positive cells were observed in the BGH group while Caspase-3 positive cells were significantly decreased in the BGH group compared with the BH group (P<0.05). Ghrelin probably enhances BMSCs viability through regulation of pro- and antiapoptotic genes Caspase 3 and Bcl2. However the signaling pathway of this effect should be elucidated in the future.
Insights
Ghrelin enhances bone marrow stromal cell (BMSC) viability by regulating apoptosis. It decreases Caspase-3 and increases Bcl-2 gene expression, suggesting a protective mechanism against cell death.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Ghrelin exhibits antiapoptotic effects in various cell types, including bone marrow stromal cells (BMSCs).
- The precise molecular mechanisms underlying ghrelin's antiapoptotic action remain unclear.
- Caspase-3 and Bcl-2 are key regulators of apoptosis in eukaryotic cells.
Purpose of the Study:
- To investigate the effect of ghrelin on the expression of Caspase-3 and Bcl-2 in BMSCs.
- To elucidate the role of ghrelin in regulating apoptotic pathways in BMSCs.
Main Methods:
- Rat BMSCs were cultured and treated with ghrelin (100 μM) and/or hydrogen peroxide (H2O2, 125 µM).
- Gene expression of Caspase-3 and Bcl-2 was analyzed using real-time PCR.
- Protein expression and localization of Caspase-3 and Bcl-2 were assessed via immunofluorescence.
Main Results:
- Ghrelin significantly reduced Caspase-3 mRNA expression compared to H2O2-treated cells (P<0.05).
- Bcl-2 gene expression increased in ghrelin-treated groups compared to H2O2-treated groups (P<0.05).
- Immunofluorescence revealed increased Bcl-2 and decreased Caspase-3 positive cells in ghrelin-treated groups.
Conclusions:
- Ghrelin likely promotes BMSC viability by modulating the expression of pro-apoptotic (Caspase-3) and anti-apoptotic (Bcl-2) genes.
- Further research is needed to fully elucidate the signaling pathways involved in ghrelin's protective effects.
