Protective effect of recombinant Lactobacillus plantarum against H2O2-induced oxidative stress in HUVEC cells
- 13 May 2021
- journal article
- research article
- Published by Zhejiang University Press in Journal of Zhejiang University-SCIENCE B
- Vol. 22 (5), 348-365
- https://doi.org/10.1631/jzus.b2000441
Abstract
This study probed the protective effect of recombinant Lactobacillus plantarum against hydrogen peroxide (H2O2)-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 H2O2-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 H2O2-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 H2O2-induced injury in HUVEC cells. The mechanism for this effect is related to enhancement of antioxidant capacity and apoptosis.Keywords
Funding Information
- National Natural Science Foundation of China (31672528, 31941018, 32072888)
- the national key Research and Development Program of China (2017YFD0501000, 2017YFD0501200)
- the Science and Technology Project of Jilin Provincial Department of Education (JJKH20190942KJ)
- the Science and Technology Development Program of Jilin Province , China (20180201040NY, 20190301042NY)
This publication has 51 references indexed in Scilit:
- Hypocaloric diet supplemented with probiotic cheese improves body mass index and blood pressure indices of obese hypertensive patients - a randomized double-blind placebo-controlled pilot studyNutrition Journal, 2013
- MicroRNAs in Stress Signaling and Human DiseaseCell, 2012
- Antiproliferative and anticytotoxic effects of cell fractions and exopolysaccharides from Lactobacillus casei 01Mutation Research - Genetic Toxicology and Environmental Mutagenesis, 2011
- β-Amyloid peptide increases levels of iron content and oxidative stress in human cell and Caenorhabditis elegans models of Alzheimer diseaseFree Radical Biology & Medicine, 2011
- Role of angiotensin II in liver fibrosis‐induced portal hypertension and therapeutic implicationsHepatology Research, 2010
- Mitochondria, oxidative stress and cell deathApoptosis, 2007
- Carvedilol protected diabetic rat hearts via reducing oxidative stressJournal of Zhejiang University-SCIENCE B, 2006
- Role of Caspases in Ox-LDL–Induced Apoptotic Cascade in Human Coronary Artery Endothelial CellsCirculation Research, 2004
- Comparison of the effects of omapatrilat and irbesartan/hydrochlorothiazide on endothelial function and cardiac hypertrophy in the stroke-prone spontaneously hypertensive ratJournal of Hypertension, 2004
- Effects of PDGF-BB on Rat Dermal Fibroblast Behavior in Mechanically Stressed and Unstressed Collagen and Fibrin GelsExperimental Cell Research, 2001