Beta-nicotinamide adenine dinucleotide (β-NAD) reverses the LPS-induced endothelial barrier dysfunction in sickle cell mice

4^th International Conference and Exhibition on Lung & Respiratory Care

August 01-02, 2016 Manchester, UK

Nagavedi Siddaramappa Umapathy

Georgia Health Sciences University, USA

Posters & Accepted Abstracts: J Pulm Respir Med

Abstract :

Background: Recent studies have demonstrated that sickle cell disease (SCD) is characterized by weakening of the endothelial barrier, which predisposes the lung to acute loss of barrier function, reminiscent of acute chest syndrome (ACS) with high morbidity and mortality. Our previous studies demonstrated the protection of �²-NAD against LPS-induced EC barrier dysfunction in an acute lung injury. However, the potential protective role of �²-NAD in ACS has not been explored. The purpose of the present study was to determine the protective effects of �²-NAD against LPS-induced EC barrier dysfunction in freshly isolated EC from lungs SCD mice. Methods: Freshly isolated endothelial cells from lungs of transgenic sickle cell mice were used in this study. The trans-endothelial electrical resistance (TER) was carried out using an electrical cell-substrate impedance sensing (ECIS) instrument. Immunofluorescence studies and Western immunoblotting were done according to standard laboratory protocol. All the reagents were obtained from Sigma unless otherwise stated. The �²-NAD was purchased from Calbiochem. Results: Our preliminary results suggest that EC isolated from sickle cell mice are more susceptible to LPS (100ng/ml) compared to their heterozygote littermates based on the TER analysis using ECIS. We observed a strikingly enhanced barrier function with �²-NAD (100 �¼M) in EC from sickle cell mice and the heterozygotes. In addition, �²-NAD significantly attenuates the LPS response as evidenced by attenuated actin stress fibers and VE-cadherin at the cell-cell junctions. Conclusions: Our data suggests that EC from sickle cell mice are more susceptible to LPS and �²-NAD reverses the LPS- induced EC barrier dysfunction.

Biography :

Email: usiddaramappa@gru.edu