Home
|
Inbox
|
Search
|
View Abstract
Effect of Advanced Glycation End Products Macrophages Under Hyperglycemic Concentrations
Description
.abstract img { width:300px !important; height:auto; display:block; text-align:center; margin-top:10px } .abstract { overflow-x:scroll } .abstract table { width:100%; display:block; border:hidden; border-collapse: collapse; margin-top:10px } .abstract td, th { border-top: 1px solid #ddd; padding: 4px 8px; } .abstract tbody tr:nth-child(even) td { background-color: #efefef; } .abstract a { overflow-wrap: break-word; word-wrap: break-word; }
A2990 - Effect of Advanced Glycation End Products Macrophages Under Hyperglycemic Concentrations
Author Block: A. Linderholm1, C. Mendez1, E. Ogorodnik2, G. Liu3, N. J. Kenyon4; 1IM-Pulmonary, Critical Care, Sleep Medicine, UC Davis, Davis, CA, United States, 2Biophysics Graduate Group, UC Davis, Davis, CA, United States, 3Chemistry, UC Davis, Davis, CA, United States, 4IM-Pulmonary, Critical Care, Sleep Medicine, UC Davis, Sacramento, CA, United States.
RATIONALE: There is strong evidence that obesity aggregates asthma symptoms through multiple inflammatory pathways. One potential pathway is glycosylated proteins and lipids, advanced glycation end products (AGEs), stimulating the transmembrane receptor, receptor for advanced glycation endproducts (RAGE), expressed in various tissues, including the lung. RAGE has been shown to activate NF-kB which regulates immune response. An abundance of macrophages are present in the lung and their activity is an important component in chronic inflammatory diseases such as asthma. This investigation aims at revealing how AGEs may alter the macrophages’ structural dynamics and if that is related to their activity. The outcomes potentially could lead to another metric in quantifying the dysregulation of immune response in the background of obesity. Using a technology known as single cell compression (SCC), the living macrophages were compressed individually in vitro via an atomic force microscopy (AFM) probe. The force versus deformation during compression and probe-cell adhesion during retreat were recorded to measure the cellular mechanics change due to macrophages AGEs interactions at low and high sugar conditions. We correlated these measurements with changes in gene expression to confirm that AGEs are altering immune cell dynamics. Our hypothesis is that AGEs induces changes in macrophages’ plasticity which potentially leads to alterations in function. METHODS: RAW 264.7 macrophages were incubated in both low (1g/L) and high (4.5g/L) glucose concentrations of DMEM for 24 hours at 37°C 5% CO2. Two AGEs concentrations (100µg/mL and 500µg/mL were tested at each glucose concentration with 500ug/mL of BSA as control. SCC and qPCR for GAPDH, TNFα and ARG1 was then performed. RESULTS: Both SCC measurements and TNFα and ARG1 expression demonstrate that exposure to the different glucose concentrations alone exhibit very similar mechanics. However, when AGEs were added, macrophage responses are identified at the higher glucose concentration. CONCLUSION: Our results suggest that the higher glucose concentration primes macrophages to respond to stimulus despite not generating a strong response alone. This suggests that diabetic conditions potentiate the immune system response.
Author Block: A. Linderholm1, C. Mendez1, E. Ogorodnik2, G. Liu3, N. J. Kenyon4; 1IM-Pulmonary, Critical Care, Sleep Medicine, UC Davis, Davis, CA, United States, 2Biophysics Graduate Group, UC Davis, Davis, CA, United States, 3Chemistry, UC Davis, Davis, CA, United States, 4IM-Pulmonary, Critical Care, Sleep Medicine, UC Davis, Sacramento, CA, United States.
RATIONALE: There is strong evidence that obesity aggregates asthma symptoms through multiple inflammatory pathways. One potential pathway is glycosylated proteins and lipids, advanced glycation end products (AGEs), stimulating the transmembrane receptor, receptor for advanced glycation endproducts (RAGE), expressed in various tissues, including the lung. RAGE has been shown to activate NF-kB which regulates immune response. An abundance of macrophages are present in the lung and their activity is an important component in chronic inflammatory diseases such as asthma. This investigation aims at revealing how AGEs may alter the macrophages’ structural dynamics and if that is related to their activity. The outcomes potentially could lead to another metric in quantifying the dysregulation of immune response in the background of obesity. Using a technology known as single cell compression (SCC), the living macrophages were compressed individually in vitro via an atomic force microscopy (AFM) probe. The force versus deformation during compression and probe-cell adhesion during retreat were recorded to measure the cellular mechanics change due to macrophages AGEs interactions at low and high sugar conditions. We correlated these measurements with changes in gene expression to confirm that AGEs are altering immune cell dynamics. Our hypothesis is that AGEs induces changes in macrophages’ plasticity which potentially leads to alterations in function. METHODS: RAW 264.7 macrophages were incubated in both low (1g/L) and high (4.5g/L) glucose concentrations of DMEM for 24 hours at 37°C 5% CO2. Two AGEs concentrations (100µg/mL and 500µg/mL were tested at each glucose concentration with 500ug/mL of BSA as control. SCC and qPCR for GAPDH, TNFα and ARG1 was then performed. RESULTS: Both SCC measurements and TNFα and ARG1 expression demonstrate that exposure to the different glucose concentrations alone exhibit very similar mechanics. However, when AGEs were added, macrophage responses are identified at the higher glucose concentration. CONCLUSION: Our results suggest that the higher glucose concentration primes macrophages to respond to stimulus despite not generating a strong response alone. This suggests that diabetic conditions potentiate the immune system response.
|
Home
|
Inbox
|
Search
|