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Protective effect of recombinant Lactobacillus plantarum against H 2 O 2 -induced oxidative stress in HUVEC cells.
- Source :
-
Journal of Zhejiang University. Science. B [J Zhejiang Univ Sci B] 2021 May 15; Vol. 22 (5), pp. 348-365. - Publication Year :
- 2021
-
Abstract
- This study probed the protective effect of recombinant Lactobacillus plantarum against hydrogen peroxide (H <subscript>2</subscript> O <subscript>2</subscript> )-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). We constructed a new functional L. plantarum (NC8-pSIP409-alr-angiotensin-converting enzyme inhibitory peptide (ACEIP)) with a double-gene-labeled non-resistant screen as an expression vector. A 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) colorimetric assay was carried out to determine the cell viability of HUVEC cells following pretreatment with NC8-pSIP409-alr-ACEIP. Flow cytometry (FCM) was used to determine the apoptosis rate of HUVEC cells. Cysteinyl aspartate specific proteinase (caspase)-3/8/9 activity was also assayed and western blotting was used to determine protein expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), inducible nitric oxide synthase (iNOS), nicotinamide adenine dinucleotide phosphate oxidase 2 (gp91phox), angiotensin II (AngII), and angiotensin-converting enzyme 2 (ACE2), as well as corresponding indicators of oxidative stress, such as reactive oxygen species (ROS), mitochondrial membrane potential (MMP), malondialdehyde (MDA), and superoxide dismutase (SOD). NC8-pSIP409-alr-ACEIP attenuated H <subscript>2</subscript> O <subscript>2</subscript> -induced cell death, as determined by the MTT assay. NC8-pSIP409-alr-ACEIP reduced apoptosis of HUVEC cells by FCM. In addition, compared to the positive control, the oxidative stress index of the H <subscript>2</subscript> O <subscript>2</subscript> -induced HUVEC (Hy-HUVEC), which was pretreated by NC8-pSIP409-alr-ACEIP, iNOS, gp91phox, MDA, and ROS, was decreased obviously; SOD expression level was increased; caspase-3 or -9 was decreased, but caspase-8 did not change; Bcl-2/Bax ratio was increased; permeability changes of mitochondria were inhibited; and loss of transmembrane potential was prevented. Expression of the hypertension-related protein (AngII protein) in HUVEC cells protected by NC8-pSIP409-alr-ACEIP decreased and expression of ACE2 protein increased. These plantarum results suggested that NC8-pSIP409-alr-ACEIP protects against H <subscript>2</subscript> O <subscript>2</subscript> -induced injury in HUVEC cells. The mechanism for this effect is related to enhancement of antioxidant capacity and apoptosis.
- Subjects :
- Angiotensin-Converting Enzyme 2 analysis
Angiotensin-Converting Enzyme Inhibitors metabolism
Apoptosis drug effects
Cells, Cultured
Human Umbilical Vein Endothelial Cells drug effects
Human Umbilical Vein Endothelial Cells metabolism
Humans
Membrane Potential, Mitochondrial drug effects
Reactive Oxygen Species metabolism
Recombinant Fusion Proteins biosynthesis
Antioxidants pharmacology
Hydrogen Peroxide pharmacology
Lactobacillus plantarum
Oxidative Stress drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1862-1783
- Volume :
- 22
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of Zhejiang University. Science. B
- Publication Type :
- Academic Journal
- Accession number :
- 33973418
- Full Text :
- https://doi.org/10.1631/jzus.B2000441