Alpha-1 Antitrypsin Inhibits Senescence in Neural Cells
School Name
Academic Magnet High School
Grade Level
12th Grade
Presentation Topic
Cell and Molecular Biology
Presentation Type
Mentored
Abstract
One in three people above the age of 65 dies from Alzheimer's disease or another form of dementia in the United States. High levels of oxidative stress have been demonstrated to induce neuron cell damage through increasing cell senescence, a common feature in Alzheimer's disease. Alpha-1 antitrypsin (AAT) is a proteinase inhibitor that plays a critical role in maintaining normal lung function, with additional anti-inflammatory and anti-apoptotic effects. The goal of this study is to test whether AAT can suppress oxidative stress-induced neuron cell death via the regulation of cellular senescence facilitated through the P21/P16 cell pathways. Samples of the neuroblastoma cell line, SH-SY5Y (ATCC), were cultured in DMEM with 10% fetal bovine serum and 1% antibiotics in 5% CO2 atmosphere. Cells (0.1x106) were seeded in 6-well plates treated with 100 μM of H2O2 to induce oxidative stress. Control cells were cultured in media without H2O2. In some groups, cells were pre-treated with different concentrations (0.1-0.5 mg/ml, Grifols) of AAT for 2 hours before being stimulated with H2O2. Cells were collected 24 hours after treatment. Cell morphology was observed. Cell deaths were measured by counting the number of surviving (attaching) cells divided by the total number of control cells. The presence of senescence cells was identified by acidic senescence-associated-β-galactosidase (SA-β-gal) staining. Cells from 5 random areas were selected and counted. Each experiment was repeated 3 times with triplicates in each sample. Differences were compared using the ANOVA test. Cells treated with H2O2 showed obvious signs of cell shrinkage, rounding, or elongation compared to untreated healthy control cells which were oval-shaped and spreading to each other. In contrast, AAT pre-treated cells showed similar morphology as untreated cells, suggesting less damage. Protein samples have been collected to analyze if AAT changes the cell cycle via flow cytometry and western blot analysis.
Recommended Citation
Lee, Michael, "Alpha-1 Antitrypsin Inhibits Senescence in Neural Cells" (2024). South Carolina Junior Academy of Science. 398.
https://scholarexchange.furman.edu/scjas/2024/all/398
Location
RITA 271
Start Date
3-23-2024 11:15 AM
Presentation Format
Oral Only
Group Project
No
Alpha-1 Antitrypsin Inhibits Senescence in Neural Cells
RITA 271
One in three people above the age of 65 dies from Alzheimer's disease or another form of dementia in the United States. High levels of oxidative stress have been demonstrated to induce neuron cell damage through increasing cell senescence, a common feature in Alzheimer's disease. Alpha-1 antitrypsin (AAT) is a proteinase inhibitor that plays a critical role in maintaining normal lung function, with additional anti-inflammatory and anti-apoptotic effects. The goal of this study is to test whether AAT can suppress oxidative stress-induced neuron cell death via the regulation of cellular senescence facilitated through the P21/P16 cell pathways. Samples of the neuroblastoma cell line, SH-SY5Y (ATCC), were cultured in DMEM with 10% fetal bovine serum and 1% antibiotics in 5% CO2 atmosphere. Cells (0.1x106) were seeded in 6-well plates treated with 100 μM of H2O2 to induce oxidative stress. Control cells were cultured in media without H2O2. In some groups, cells were pre-treated with different concentrations (0.1-0.5 mg/ml, Grifols) of AAT for 2 hours before being stimulated with H2O2. Cells were collected 24 hours after treatment. Cell morphology was observed. Cell deaths were measured by counting the number of surviving (attaching) cells divided by the total number of control cells. The presence of senescence cells was identified by acidic senescence-associated-β-galactosidase (SA-β-gal) staining. Cells from 5 random areas were selected and counted. Each experiment was repeated 3 times with triplicates in each sample. Differences were compared using the ANOVA test. Cells treated with H2O2 showed obvious signs of cell shrinkage, rounding, or elongation compared to untreated healthy control cells which were oval-shaped and spreading to each other. In contrast, AAT pre-treated cells showed similar morphology as untreated cells, suggesting less damage. Protein samples have been collected to analyze if AAT changes the cell cycle via flow cytometry and western blot analysis.
