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l-Cysteine in vitro can restore cellular glutathione and inhibits the expression of cell adhesion molecules in G6PD-deficient monocytes

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Abstract

l-Cysteine is a precursor of glutathione (GSH), a potent physiological antioxidant. Excess glucose-6-phosphate dehydrogenase (G6PD) deficiency in African Americans and low levels of l-cysteine diet in Hispanics can contributes to GSH deficiency and oxidative stress. Oxidative stress and monocyte adhesion was considered to be an initial event in the progression of vascular dysfunction and atherosclerosis. However, no previous study has investigated the contribution of GSH/G6PD deficiency to the expression of monocyte adhesion molecules. Using human U937 monocytes, this study examined the effect of GSH/G6PD deficiency and l-cysteine supplementation on monocyte adhesion molecules. G6PD/GSH deficiency induced by either siRNA or inhibitors (6AN/BSO, respectively) significantly (p < 0.005) increased the levels of cell adhesion molecules (ICAM-1, VCAM-1, SELL, ITGB1 and 2); NADPH oxidase (NOX), reactive oxygen species (ROS) and MCP-1 were upregulated, and decreases in levels of GSH, and nitric oxide were observed. The expression of ICAM-1 and VCAM-1 mRNA levels increased in high glucose, MCP-1 or TNF-α-treated G6PD-deficient compared to G6PD-normal cells. l-Cysteine treatment significantly (p < 0.005) increased G6PD activity and levels of GSH, and decreased NOX, ROS, and adhesion molecules. Thus, GSH/G6PD deficiency increases susceptibility to monocyte adhesion processes, whereas l-cysteine supplementation can restore cellular GSH/G6PD and attenuates NOX activity and expression of cell adhesion molecules.

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Abbreviations

6-AN:

6-Aminonicotinamide

AA:

African American

BCA:

Bicinchoninic acid

BSO:

Buthionine sulfoximine

CAM:

Cell adhesion molecules

CCL2:

Chemokine (C–C motif) ligand 2

CVD:

Cardiovascular disease

G6PD:

Glucose-6-phosphate dehydrogenase

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GCLC:

Glutamate–cysteine ligase catalytic subunit

GCLM:

Glutamate–cysteine ligase modifier subunit

GSH:

Glutathione

GSS:

Glutathione synthetase

HG:

High glucose

ICAM-1:

Intercellular adhesion molecule 1

ITGB:

Integrin subunit beta

KD:

Knockdown

MCP-1:

Monocyte chemoattractant protein 1

NO:

Nitric oxide

NOX:

NADPH oxidase

ROS:

Reactive oxygen species

SELL:

Selectin L

siRNA:

Short interference RNA

SLC7A11:

Solute carrier family 7 member

TNF-α:

Tumor necrosis factor-α

VCAM-1:

Vascular cell adhesion protein 1

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Acknowledgements

The authors are supported by grants from the Malcolm W. Feist Cardiovascular Research Fellowship to Rajesh Parsanathan and Endowed Chair in Diabetes to Sushil K. Jain from the Center for Cardiovascular Diseases and Sciences (CCDS), LSUHSC, Shreveport. We also thank the Research Core Facility at Louisiana State University Health Sciences Center in Shreveport for generation of the gene expression data. Ms. Paula Polk kindly assisted with the 384-well block installation in the Applied Biosystems 7900HT Fast Real-Time PCR System. The authors thank Ms. Georgia Morgan for excellent editing.

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  1. Department of Pediatrics and Center for Cardiovascular Diseases and Sciences, Louisiana State University Health Sciences Center-Shreveport, 1501 Kings Highway, Shreveport, LA, 71130, USA

    Rajesh Parsanathan & Sushil K. Jain

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  1. Rajesh Parsanathan
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Correspondence to Sushil K. Jain.

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Parsanathan, R., Jain, S.K. l-Cysteine in vitro can restore cellular glutathione and inhibits the expression of cell adhesion molecules in G6PD-deficient monocytes. Amino Acids 50, 909–921 (2018). https://doi.org/10.1007/s00726-018-2559-x

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