Society of Black Academic SurgeonsAlpha lipoic acid attenuates microvascular endothelial cell hyperpermeability by inhibiting the intrinsic apoptotic signaling
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Chemicals and solutions
ALA and fluorescein isothiocyanate-bovine albumin (FITC-albumin) were obtained from Sigma (St. Louis, MO). JC-1 was obtained from Cell Technology Inc. (Mountain View, CA). JC-1 was prepared by reconstituting the lyophilized reagent with 500 μL of dimethyl sulfoxide to obtain a 100× stock solution. Immediately before the experiments the 100× solution was diluted 1:100 in 1× assay buffer. BAK (BH3) peptide and BAK L to A mutant peptide (R&D Systems, Minneapolis, MN), a 1 μg/μL stock, was mixed
Monolayer hyperpermeability
Transfection of BAK (BH3) peptide (5 μg/mL) on RLMEC monolayers induced hyperpermeability. The FITC-albumin fluorescent intensity of the control cells was taken as 100%. The FITC-albumin fluorescent intensity was significant in the BAK (BH3) peptide–transfected group compared with the control group (215.4% ± 8.4% of the control; P < .05; Fig. 1A), suggesting leakage of albumin across the endothelial monolayer. Cells pretreated with ALA (100 μmol/L) showed a decrease in BAK (BH3)-induced
Comments
In this study, we tested the ability of ALA, an endogenous antioxidant with anti-apoptotic properties, to attenuate microvascular endothelial cell hyperpermeability. BAK, a pro-apoptotic factor, was used to induce hyperpermeability. Our results showed that BAK (BH3) transfection increased microvascular endothelial cell hyperpermeability, which was attenuated by ALA pretreatment. In addition, ALA inhibited BAK (BH3)-induced mitochondrial ROS formation, prevented the collapse of mitochondrial
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